Дисертації з теми "Tortricid pests"

Thesis (MScAgric)--Stellenbosch University, 2013.
ENGLISH ABSTRACT: Cydia pomonella (codling moth), Thaumatotibia leucotreta (False codling moth), Thaumatotibia batrachopa (Macadamia nut borer), Grapholita molesta (Oriental fruit moth), Cryptophlebia peltastica (Litchi moth), Epichoristodes acerbella (Pear leafroller/Carnation worm) and Lozotaenia capensana (Apple leafroller) are the most economically important tortricids affecting various crops in South Africa. The correct identification of these species, especially of the larval stage, is of great importance in pest management. Using available literature, augmented by additional morphological studies, an interactive identification key (Lucid key) for larval and adult stages of the seven species was developed. The colour and markings of the head, characteristics of the prothoracic and anal shields, the position of the prespiracular setae (L-group) relative to the spiracle on the prothoracic segment, the position of the spiracle on the eighth abdominal segment and L-group on the ninth abdominal segment, as well as the presence or absence of the anal comb are key characteristics for larval identification. For adult identification, wing pattern and genitalia are the most important features. However, the use of genitalia for moth identification might be difficult for the lay user, as the dissection and mounting of these structures requires certain skills and specialized equipment. Thus, genitalia have not been included in the Lucid Key. Differences in the morphological characteristics of most pupae were so minute that this stage was also not included in the Lucid key. However, the pupae of E. acerbella and L. capensana are easily distinguished from those of the other species by the presence of acremaster. This study also included the first morphological description of the pupa of L. capensana, which can be distinguished from that of E. acerbella by various features of the cremaster, antennae, spiracle shape, number of setae on abdominal segments A5-7, the size of spines on A3-7, and the presence/absence of spines on A9. A previous study by Timm (2005) indicated that geographically isolated populations of T. leucotreta tend to be genetically distinct. This raised the question of whether speciation/subspeciation has occurred or is occurring. Male moth genitalia are thought to evolve rapidly and are often the only features that can reliably distinguish similar species. Hence, variation in the shape of the valvae of T. leucotreta was used to determine whether divergence has occurred between populations of T. leucotreta. Elliptical Fourier analysis was used to analyze the valvar variation in three different populations. Although some variation in valvar shape was detected among mean population values for certain traits, no clear pattern emerged. Principle component analysis also showed no distinct clustering of valvae shape among populations, providing no evidence for divergence in male genitalia and therefore no morphological evidence of incipient speciation.
AFRIKAANSE OPSOMMING: Cydia pomonella (Kodlingmot), Thaumatotibia leucotreta (Valskodlingmot), T. batrachopa (Makadamianeutboorder), Grapholita molesta (Oosterse vrugtemot), Cryptophlebia peltastica (Lietsjiemot), Epichoristodes acerbella (Peerbladroller/Angelierrusper) en Lozotaenia capensana (Appelbladroller) is die mees ekonomies belangrike tortrisiede van die vrugtebedryf in Suid-Afrika. Die juiste identifikasie van hierdie spesies, veral van hulle larwale stadium, is van groot belang by plaagbestuur. Deur gebruik te maak van beskikbare literatuur, aangevul deur bykomstige morfologiese studies, is ‗n interaktiewe uitkenningssleutel (―Lucid key‖) vir die larwale en volwasse stadia van die sewe spesies ontwikkel. Die kleur en tekening van die kop, kenmerke van die prothorakale en anale skild, die ligging van die prespirakulêre setae (L-groep) relatief tot die spiraculum op die prothorakale segment, die ligging van die spirakulum op die agste abdominale segment en L-groep op die negende abdominale segment, asook die aan- of afwesigheid van die anale kam is sleutel kenmerke vir larwale uitkenning. Vir die volwassenes is die vlerktekening en genitalia die mees belangrike kenmerke. Die gebruik van die genitalia vir motuitkenning kan egter vir die leek gebruiker moeilik wees omdat die disseksie en montering van hierdie strukture bepaalde vaardighede en gespesialiseerde toerusting vereis. Vir die rede is die genitalia nie in die Lucid-sleutel ingesluit nie. Verskille in die morfologiese kenmerke van meeste papies is klein en die stadium is gevolglik ook nie in die sleutel ingesluit nie. Die papies van E. acerbella en L. capensana kan egter maklik van die ander spesies onderskei word deur die aanwesigheid van ‗n cremaster. Hierdie studie sluit ook die eerste morfologiese beskrywing van die papie van L. capensana in, wat van dié van E. acerbella onderskei kan word deur gebruik te maak van kenmerke van die cremaster, antennae, spirakulêre vorm, aantal setae op abdominale segmente A5-7, die grootte van stekels op A3-7, en die aan- of afwesigheid van stekels op A9. ‗n Vroeëre studie (Timm 2005) het aangedui dat geografies geïsoleerde bevolkings van T. leucotreta neig om geneties verskillend te wees. Dit het die vraag laat ontstaan of spesiasie/subspesiasie moontlik plaasgevind het of steeds plaasvind. Manlike mot genitalië word geag om vinnig te ontwikkel en is dikwels die enigste kenmerke wat betroubaar tussen soortgelyke spesies kan onderskei. Dus is die variasie in die vorm van die valvae van T. leucotreta gebruik om te bepaal of divergensie wel tussen bevolkings van T. leucotreta plaasgevind het. Elliptiese Fourier ontleding is gebruik om die valvae se variasie by drie verskillende bevolkings te ontleed. Alhoewel enkele variasie in die vorm van die valvae bespeur is by die gemiddelde bevolkingswaardes vir bepaalde eienskappe, kon geen duidelike patroon bespeur word nie. Hoofkomponentontleding het ook geen duidelike groepering van valvae se vorm tussen bevolkings getoon nie, wat geen bewys lewer van divergensie in die manlike genitalia en dus geen morfologiese bewys van beginnende spesiasie.

Pest control strategies targeting insect olfaction represent a promising venue for control of tortricid insects (Lepidoptera: Tortricidae). Among tortricids, the grapevine moth Lobesia botrana (Denis and Schiffermüller) and the codling moth Cydia pomonella (L.) are serious pests for worldwide production of fruit crops. We employed several approaches to the olfactory system, from electrophysiological and behavioral studies in the grapevine moth, to bioinformatic and molecular studies of olfactory sensory proteins in the codling moth. At the receptor level, we studied both the Olfactory Receptors (ORs), the most common class of sensory proteins mediating detection of odors in insect antennae, and the Transient Receptor Potential (TRP) channels, a novel family of receptor, that recently were also found in the antennae of lepidopterous species. We demonstrated electrophysiological and behavioral responses of the grapevine moth to volatiles emitted by a non-host, Perilla frutescens, previously known to activate TRPs in the rat, Rattus norvegicus. In the codling moth, we characterized a novel TRP channel (TRPA pyrexia-like) and we confirmed activation of its human orthologue to the same non-host compounds active on the olfactory system of the grapevine moth. ORs were heterologously expressed in vivo and in vitro, for identification of their ligands among host and non-host plant volatiles and pheromones (deorphanization). Among several ORs of codling moth, we deorphanized a candidate pheromone receptor (PR) to plant synergists, an OR to non-host volatiles and another PR candidate to a pheromone antagonist of the insect. Our study thus opens for refinement of existing pest control, or novel applications. The behavioral response of the grapevine moth to volatiles from a nonhost plant, and the identification of a novel TRP channel in the codling moth may have perspectives for application in agriculture, targeting the somatosensory system of these tortricids. The evolutionary implications of the responses of the human orthologue of TRPA pyrexia-like to volatiles active on the grapevine moth olfactory system could imply a large degree of conservation of the receptor function. In the codling moth, identification of synergist and antagonist ligands for candidate PRs and deorphanization of an OR to non-host plant volatiles suggest a possible role of these receptors in reproductive and ecological isolation. This could lead to further refinement of existing semiochemicalbased control techniques, by enabling a better understanding of mate- and host-finding in this species.

Pest control strategies targeting insect olfaction represent a promising venue for control of tortricid insects (Lepidoptera: Tortricidae). Among tortricids, the grapevine moth Lobesia botrana (Denis and Schiffermüller) and the codling moth Cydia pomonella (L.) are serious pests for worldwide production of fruit crops. We employed several approaches to the olfactory system, from electrophysiological and behavioral studies in the grapevine moth, to bioinformatic and molecular studies of olfactory sensory proteins in the codling moth. At the receptor level, we studied both the Olfactory Receptors (ORs), the most common class of sensory proteins mediating detection of odors in insect antennae, and the Transient Receptor Potential (TRP) channels, a novel family of receptor, that recently were also found in the antennae of lepidopterous species. We demonstrated electrophysiological and behavioral responses of the grapevine moth to volatiles emitted by a non-host, Perilla frutescens, previously known to activate TRPs in the rat, Rattus norvegicus. In the codling moth, we characterized a novel TRP channel (TRPA pyrexia-like) and we confirmed activation of its human orthologue to the same non-host compounds active on the olfactory system of the grapevine moth. ORs were heterologously expressed in vivo and in vitro, for identification of their ligands among host and non-host plant volatiles and pheromones (deorphanization). Among several ORs of codling moth, we deorphanized a candidate pheromone receptor (PR) to plant synergists, an OR to non-host volatiles and another PR candidate to a pheromone antagonist of the insect. Our study thus opens for refinement of existing pest control, or novel applications. The behavioral response of the grapevine moth to volatiles from a nonhost plant, and the identification of a novel TRP channel in the codling moth may have perspectives for application in agriculture, targeting the somatosensory system of these tortricids. The evolutionary implications of the responses of the human orthologue of TRPA pyrexia-like to volatiles active on the grapevine moth olfactory system could imply a large degree of conservation of the receptor function. In the codling moth, identification of synergist and antagonist ligands for candidate PRs and deorphanization of an OR to non-host plant volatiles suggest a possible role of these receptors in reproductive and ecological isolation. This could lead to further refinement of existing semiochemicalbased control techniques, by enabling a better understanding of mate- and host-finding in this species.

False codling moth (FCM), Thaumatotibia leucotreta is an extremely important pest of citrus in South Africa and with the shift away from the use of chemicals, alternate control options are needed. One avenue of control which has only recently been investigated against the soil-borne life stages of FCM is the use of entomopathogenic fungi (EPF). In 2009, 12 entomopathogenic fungal isolates collected from South African citrus orchards showed good control potential during laboratory conducted bioassays. The aim of this study was to further analyse the potential of these isolates through concentration-dose and exposure-time response bioassays. After initial re-screening, concentration-dose response and exposure-time response sandconidial bioassays, three isolates were identified as exhibiting the greatest control potential against FCM in soil, Metarhizium anisopliae var. anisopliae (G 11 3 L6 and FCM Ar 23 B3) and Beauveria bassiana (G Ar 17 B3). Percentage mycosis was found to be directly related to fungal concentration as well as the amount of time FCM 5th instar larvae were exposed to the fungal conidia. LC50 values for the three isolates were not greater than 1.92 x 10⁶ conidia.ml⁻ₑ and at the LC₅₀, FCM 5th instar larvae would need to be exposed to the fungus for a maximum of 13 days to ensure a high mortality level. These isolates along with two commercially available EPF products were subjected to field persistence trials whereby net bags filled with a mixture of autoclaved sand and formulated fungal product were buried in an Eastern Cape citrus orchard. The viability of each isolate was measured on a monthly basis for a period of six months. All isolates were capable of persisting in the soil for six months with the collected isolates persisting far better than the commercially used isolates. Two of the isolates, G 11 3 L6 and G Ar 17 B3, were subjected to small scale laboratory application trials. Two formulations were investigated at two concentrations. For each isolate, each formulation and each concentration, FCM 5th instar larvae were applied and allowed to burrow into the soil to pupate before fungal application or after fungal application. Contact between fungi and FCM host is essential as, in contrast to pre-larval treatments, percentage mortality in post-larval treatments was low for both formulations and both isolates. For isolate G Ar 17 B3, a conidial suspension applied as a spray at a concentration of 1 x 10⁷ conidia.ml⁻ₑ obtained the highest percentage mortality (80 %). For isolate G 11 3 L6 however, both formulations performed equally well at a high, 1 x10⁷ conidia.ml⁻ₑ concentration (conidial suspension: 60 %; granular: 65 %) The results obtained thus far are promising for the control of FCM in citrus, but if these EPFs are to successfully integrate into current FCM control practices more research, some of which is discussed, is essential

Control of the citrus pest, false codling moth (FCM), Thaumatotibia leucotreta Meyrick (Lepidoptera: Tortricidae) is crucial for the South African citrus industry. The economic losses and phytosanitary status of this pest, coupled with increased consumer awareness and demands, has created a need for effective, IPM-compatible control measures for use against the soil-dwelling life stages of FCM. Promising developments in the field of microbial control through the use of entomopathogenic fungi (EPF) and entomopathogenic nematodes (EPNs) have highlighted the need for research regarding pupation biology, behaviour and survival of FCM, as a good understanding of biology of the target organism is an important component of any biological control programme. The aim of this study was to improve the current understanding of FCM pupation habits through the manipulation of soil texture class, ground cover, shading, soil compaction, air temperature, and soil moisture in the laboratory. These findings would then be used to aid the biological control programmes using EPF and EPNs against FCM in the soil. Three soil texture classes (sandy loam, silt loam and silty clay loam) were obtained from orchards for use in the study. FCM larvae were allowed to drop into the soil of their own accord and the pupation behaviour that followed was then captured on film with pupae formed in the soil being kept in order to measure adult eclosion. In general, very few abiotic factors had a clear influence on FCM pupation. Larval wandering time and distance was short, but also variable between individuals. Distance did increase when soils were moist. Pupation depth was shallow, with pupal cocoons generally being formed on the soil surface. Depth of pupation was less than one centimetre for all abiotic conditions, with little burrowing into soil. Eclosion success was higher for sandier soils when these were dry and uncompacted, but the addition of both moisture and soil compaction increased FCM eclosion success. FCM was sensitive to desiccation when the soils were dry and temperature limits of 15 °C and 32 °C had a strongly negative impact on eclosion success. Preferences for particular abiotic conditions were limited to only certain moisture conditions when interacting with soil texture class and a preference for pupating in soil when it is available. Limited preference was found for particular soil textures despite this having a strong influence on eclosion success, but individuals did appear to pupate in close proximity to one another. Viable direct habitat manipulation for FCM control could not be identified. These results and all of the abiotic variables measured have important implications for EPF and EPN application, survival and persistence in the soil in order to improve the ability of these biological control agents to control FCM. These are discussed in each chapter.

The study examined the larval parasitoids of Cryptophlebia leucotreta (Meyrick) on citrus in South Africa and aimed to improve the existing rearing techniques of C. leucotreta with a view to mass rearing of biological control agents. The biological characteristics of Agathis bishopi Nixon (Hymenoptera: Braconidae) were studied, with an emphasis on parasitism rates in the field, host stage preference, developmental rate, life span and offspring sex ratios. Two larval parasitoids, A. bishopi and Apophua leucotretae (Wilkinson) (Hymenoptera: Ichneumonidae), and an egg parasitoid, Trichogrammatoidea cryptophlebiae Nagaraja (Hymenoptera: Trichogrammatidae), were recorded on C. leucotreta on citrus. A. bishopi was the more abundant of the larval parasitoids and exhibited density dependent parasitism. The highest parasitism rates were observed in December with up to 38% in Sundays River Valley and 34% in Gamtoos River Valley, at a time when the highest false codling moth infestations were observed. Agathis bishopi was recorded only in the Eastern Cape Province. The sex ratio of A. bishopi was biased towards females throughout the study (77% in Gamtoos River Valley and 72% in Sundays River Valley). Agathis bishopi is a solitary, koinobiont, larval-pupal endoparasitoid. The species showed a preference for 1st and 2"d instar hosts. Females regulate the sex of their progeny according to the size and larval stage of the host, ovipositing unfertilised eggs in younger, smaller larvae (1st instars) and fertilised eggs in older, larger larvae (2nd instars). The developmental rate of A. bishopi is in synchrony with that of the moth and adults emerge when adult moths that have escaped parasitism emerge. Agathis bishopi and T. cryptophlebiae compliment each other because they have different niches and strategies of attack. Integrating A. bishopi and T. cryptophlebiae into the management of citrus orchards has potential to suppress false codling moth. Larger rearing containers seemed ideal for large-scale rearing of false codling moth. A higher percentage of adults was obtained from larvae reared in larger containers than in smaller ones. The width of the sponges used as stoppers prevented escape of the larvae. Media prepared in larger containers are easier and simpler to prepare than in smaller ones, thus eliminating many precautions otherwise necessary to prevent contamination. Moth production was greatly reduced by the high concentration of Sporekill used for egg decontamination.

The false codling moth, Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae), is one of the major pests of citrus in South Africa, the others being mealybug, Mediterranean fruit fly, bollworm and some mites. Due to problems such as the expense of pesticides, insects evolving pesticide resistance (Hogsette 1999), chemical residue on the skin of export fruit and the negative impact of pesticides on the environment, it became necessary to find alternative methods for pest control (Viggiani 2000). Agathis bishopi (Nixon) (Hymenoptera: Braconidae), a larval parasitoid of false codling moth known only from the Sundays River Valley area (Sishuba 2003), offers a means of control for the pest. A total of 11 389 navel oranges were collected from various orchards in the Addo/Kirkwood area, and false codling moth larvae infested 36.09% of the fruit. A single parasitoid species, A. bishopi, was reared from these larvae. In 2006 the highest parasitism rate, 11.43%, was recorded in May and in 2007, the highest parasitism rate, 13.27%, was in April. Agathis bishopi parasitizes larvae in instars 2 and 3, possibly due to the accessibility of these younger instars to the female parasitoid and possibly due to the length of the life cycle of this koinobiont. Second instar hosts yielded the highest number of parasitoids, and there was no emergence of parasitoids from fifth instar larvae. Females of A. bishopi live for 18.5 days (n = 20; S.E. = 3.1) and males for 8.25 days (n = 20; S.E. = 1.23). Females produce an average of 23 offspring in a lifetime, while female false codling moths produce about 800 eggs each. A high number of parasitoids will be required per hectare to reduce the population of false codling moth. Captive rearing of A. bishopi proved difficult due to viral and fungal contamination. Agathis bishopi has potential for use in an integrated pest management programme once the hurdle of mass-rearing has been overcome.

The false codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae), is an important pest of citrus in South Africa and challenging to manage due to its inconspicuous nature. An effective method currently being employed for the area-wide suppression of the FCM is the Sterile Insect Technique (SIT) and the effective dispersal of sterile moths is very important for success with SIT. This study was conducted in the Addo area of the Sundays River Valley (Eastern Cape) where the programme is commercially used. In this study, sterile male moths were released in different orchards on a citrus farm, and in nearby veld at different times of the year, and their dispersal was monitored through the use of pheromone traps. Various climatic factors were monitored. This provided insight into the local dispersal of sterile male FCM adults in response to abiotic cues (particularly climatic factors). The movement of the FCM in four citrus cultivars, namely lemons, navel and Valencia oranges and mandarins and in the nearby veld (open field), was determined at six different stages of the year. Results clearly indicated that sterile FCM movement is concentrated within citrus orchards, as very few moths were trapped beyond 30 m from the release point, particularly in navel and Valencia orchards. Of the climatic factors measured, minimum and maximum temperatures had the most significant influence on FCM dispersal, and based on the results, various recommendations are made for the releases of sterile FCM in an area-wide SIT management programmes on citrus. A better understanding of the dispersal capabilities of the FCM in an agricultural system, under different conditions and at different times of the year, is invaluable not only in improving release strategies in an SIT programme but in planning future control strategies against the FCM.

La confusió sexual mitjançant feromones és una tàctica de control de plagues que, en impedir l'aparellament entre arnes, ha ajudat enormement a reduir els danys en cultius, així com l'ús de pesticides. La necessitat de disminuir els costos associats a la seva aplicació ha fomentat el desenvolupament de dispensadors automàtics o puffers. Aquests puffers alliberen una gran quantitat de feromona mentre els insectes estan sexualment actius. No obstant això, la falta d'informació sobre la biologia i el comportament dels insectes i de l'efecte que alguns factors externs tenen sobre la seva periodicitat diària impedeixen un correcte ajust dels puffers, la qual cosa pot suposar una disminució de la seva eficàcia en camp. A pesar que el parany automàtic pot servir per a determinar aquests períodes d'activitat i decidir quan alliberar feromona, els paranys disponibles comercialment no confereixen una resolució temporal suficient per a esbossar l'activitat dels insectes, que pot estar reduïda a unes poques hores. Per a la meva tesi hem dissenyat i posat a prova un parany assequible i fàcil de construir que m'ha permès determinar el vol sexual diari i estacional de Grapholita molesta (Busk) a partir de fotografies preses amb una gran resolució temporal. Tant en paranys encebats amb feromona sintètica com en paranys encebats amb femelles verges, la majoria de les captures van tenir lloc entre 3 hores abans i una hora després de la posta de sol, indicant una activitat diürna crepuscular en l'espècie. El tipus d'esquer (septe de feromona o femella verge) no va afectar al període de vol diari. Així i tot, les baixes temperatures van avançar els vols, mentre que les temperatures més altes retardaven el vol fins a hores pròximes a la posta de sol. Aquests resultats destaquen la necessitat de programar els puffers perquè alliberin la feromona en diferents hores al llarg de l'any, seguint la corba de vol dels insectes. Fent-t'ho així augmentaria l'eficàcia d'aquest sistema en camp, al mateix temps que reduiria el cost de la seva aplicació i evitaria un desaprofitament de feromona. Per a explicar els mecanismes subjacents al funcionament de la confusió sexual en el control de plagues, la majoria dels estudis s'han centrat en l'efecte de grans nivells de feromona sintètica sobre els mascles. Les femelles, d'altra banda, s'han considerat insensibles a la seva pròpia feromona. No obstant això, una revisió recent indica que hi ha una alteració notable en el comportament de les femelles en ser exposades a la seva pròpia feromona, un fenomen conegut com a “autodetecció”. Malgrat això, no hi ha proves que confirmin que les femelles siguin capaces de detectar la seva pròpia feromona a nivell de les neurones olfactòries. Mitjançant tècniques d'electrofisiologia, he comparat la resposta de les neurones olfactòries allotjades en sensilla trichodea de mascles i femelles a compostos biològicament rellevants per a G. molesta, una espècie en la qual s'ha descrit autodetecció. Una anàlisi d'agrupament jeràrquic indica un sistema olfactori perifèric radicalment diferent en cada sexe, la qual cosa podria estar relacionat amb les diferents necessitats biològiques de cadascun. Mentre que en els mascles no hi havia cèl·lules que responguessin a la seva pròpia feromona de festeig, cinamato d'etil, la majoria (63%) van respondre específicament als compostos de feromona sexual majoritari i minoritari (Z8-12:Ac i E8-12:Ac, respectivament). La freqüència de cadascun d'aquests tipus de cèl·lules és similar al ràtio de mescla de feromones de les femelles. En femelles, un 6% de les cèl·lules responien a volàtils de planta i un 3% són específiques per a la feromona de festeig dels mascles. A pesar que algunes cèl·lules eren estimulades per la feromona sexual de les femelles, aquestes respostes eren sempre menors que les de mascles i, a més, responien a més composts. De fet, l'agrupació jeràrquica les inclou en un grup (91%) de neurones inespecífiques. L'absència en femelles de receptors per a la feromona sexual i la seva baixa sensibilitat a la mateixa els impediria detectar variacions en les ràtios de feromona en la naturalesa. Per tant, qualsevol alteració del comportament després d'una exposició a feromona coespecífic en el laboratori no podria ser atribuïda a un reconeixement de la pròpia espècie. El comportament de les femelles sí que es pot veure alterat per altres senyals més rellevants per a les seves necessitats biològiques. L'olor característica d'una planta pot provocar un efecte diferent en femelles que provenen d'hostes diferents. En la meva tesi he analitzat respostes electroantenogràfiques de mascles i femelles adults de Lobesia botrana (Denis et Schiffermüller) (Lepidoptera: Tortricidae) recollides en la fase larvària de la vinya (Vitis vinífera L.) o del torvisco (Daphne gnidium L.) a volàtils específics i compartits dels hostes, així com a compostos de feromona sexual. Els meus resultats indiquen que la detecció de feromona no difereix entre totes dues poblacions. A més, la detecció de volàtils de planta no es veu afectada pel sexe o la planta de desenvolupament larvari. Poblacions polífagues desenvolupant-se en un hoste concret semblen retenir la capacitat per a respondre als volàtils d'altres hostes. Aquesta falta de diferències estadístiques en la diferenciació de compostos olfactoris al nivell de l'antena no implica que els individus que provenen de cada hoste mostrin preferències similars davant tots dos conjunts de volàtils, atès que les respostes biològiques depenen en última instància de la integració cerebral de cada individu. Així i tot, és important comprendre la capacitat dels insectes polífags per a reproduir-se o buscar aliments i refugis alternatius a l'hora de determinar els límits topogràfics de la confusió sexual. La meva tesi serveix per a entendre millor les repercussions que alguns factors ambientals poden tenir sobre el funcionament d'una tècnica complexa com és la confusió sexual. A pesar que l'ús de pesticides s'ha reduït, encara ens valem d'aquests químics per a evitar el descontrol de les plagues, i és necessari conèixer les variables que afecten l'eficàcia de les alternatives sostenibles.
La confusión sexual mediante feromonas es una táctica de control de plagas que, al impedir el apareamiento entre polillas, ha ayudado enormemente a reducir los daños en cultivos, así como el uso de pesticidas. La necesidad de disminuir los costes asociados a su aplicación ha fomentado el desarrollo de dispensadores automáticos o puffers. Estos puffers liberan una gran cantidad de feromona mientras los insectos están sexualmente activos. Sin embargo, la falta de información sobre la biología y el comportamiento de los insectos y del efecto que algunos factores externos tienen sobre su periodicidad diaria impiden un correcto ajuste de los puffers, lo que puede suponer una disminución de su eficacia en campo. A pesar de que el trampeo automático puede servir para determinar estos periodos de actividad y decidir cuándo liberar feromona, las trampas disponibles comercialmente no confieren una resolución temporal suficiente para bosquejar la actividad de los insectos, que puede estar reducida a unas pocas horas. Para mi tesis hemos diseñado y puesto a prueba una trampa asequible y fácil de construir que me ha permitido determinar el vuelo sexual diario y estacional de Grapholita molesta (Busk) a partir de fotografías tomadas con una gran resolución temporal. Tanto en trampas cebadas con feromona sintética como en trampas cebadas con hembras vírgenes, la mayoría de las capturas tuvieron lugar entre 3 horas antes y una hora después de la puesta de sol, indicando una actividad diurna crepuscular en la especie. El tipo de cebo (septo de feromona o hembra virgen) no afectó el periodo de vuelo diario. Aun así, las bajas temperaturas avanzaron los vuelos, mientras que las temperaturas más altas retrasaban el vuelo hasta horas cercanas a la puesta de sol. Estos resultados destacan la necesidad de programar los puffers para que liberen la feromona a distintas horas a lo largo del año, siguiendo la curva de vuelo de los insectos. Hacerlo aumentaría la eficacia de este sistema en campo, al tiempo que reduciría el coste de su aplicación y evitaría un desperdicio de feromona. Para explicar los mecanismos que subyacen al funcionamiento de la confusión sexual en el control de plagas, la mayoría de los estudios se han centrado en el efecto de grandes niveles de feromona sintética sobre los machos. Las hembras, por otro lado, se han considerado insensibles a su propia feromona. Sin embargo, una revisión reciente indica que hay una alteración notable en el comportamiento de las hembras al ser expuestas a su propia feromona, un fenómeno conocido como “autodetección”. A pesar de ello, no hay pruebas que confirmen que las hembras sean capaces de detectar su propia feromona a nivel de las neuronas olfativas. Mediante técnicas de electrofisiología, he comparado la respuesta de las neuronas olfativas alojadas en sensilla trichodea de machos y hembras a compuestos biológicamente relevantes para G. molesta, una especie en la que se ha descrito autodetección. Un análisis de agrupamiento jerárquico indica un sistema olfativo periférico radicalmente diferente en cada sexo, lo que podría estar relacionado con las diferentes necesidades biológicas de cada uno. Mientras que en los machos no había células que respondieran a su propia feromona de cortejo, cinamato de etilo, la mayoría (63%) respondieron específicamente a los compuestos de feromona sexual mayoritario y minoritario (Z8-12:Ac y E8-12:Ac, respectivamente). La frecuencia de cada uno de estos tipos de células es similar al ratio de mezcla de feromonas de las hembras. En hembras, un 6% de las células respondían a volátiles de planta y un 3% son específicas para la feromona de cortejo de los machos. A pesar de que algunas células eran estimuladas por la feromona sexual de las hembras, estas respuestas eran siempre menores que las de machos y, además, respondían a más compuestos. De hecho, la agrupación jerárquica las incluye en un grupo (91%) de neuronas inespecíficas. La ausencia en hembras de receptores para la feromona sexual y su baja sensibilidad a la misma les impediría detectar variaciones en las ratios de feromona en la naturaleza. Por lo tanto, cualquier alteración del comportamiento tras una exposición a feromona conspecífica en el laboratorio no podría ser atribuida a un reconocimiento de la propia especie. El comportamiento de las hembras sí que puede verse alterado por otras señales más relevantes para sus necesidades biológicas. El olor característico de una planta puede provocar un efecto diferente en hembras que provienen de huéspedes distintos. En mi tesis he analizado respuestas electroantenográficas de machos y hembras adultos de Lobesia botrana (Denis et Schiffermüller) (Lepidoptera: Tortricidae) recogidas en fase larvaria de la vid (Vitis vinífera L.) o del torvisco (Daphne gnidium L.) a volátiles específicos y compartidos de los huéspedes, así como a compuestos de feromona sexual. Mis resultados indican que la detección de feromona no difiere entre ambas poblaciones. Además, la detección de volátiles de planta no se ve afectada por el sexo o la planta de desarrollo larvario. Poblaciones polífagas desarrollándose en un huésped concreto parecen retener la capacidad para responder a los volátiles de otros huéspedes. Esta falta de diferencias estadísticas en la diferenciación de compuestos olfativos al nivel de la antena no implica, sin embargo, que los individuos que provienen de cada huésped muestren preferencias similares ante ambos conjuntos de volátiles, dado que las respuestas biológicas dependen en última instancia de la integración cerebral de cada individuo. Aun así, es importante comprender la capacidad de los insectos polífagos para reproducirse o buscar alimentos y refugios alternativos a la hora de determinar los límites topográficos de la confusión sexual. Mi tesis sirve para entender mejor las repercusiones que algunos factores ambientales pueden tener sobre el funcionamiento de una técnica compleja como es la confusión sexual. A pesar de que el uso de pesticidas se ha reducido, aún nos valemos de estos químicos para evitar el descontrol de las plagas, y es necesario conocer las variables que afectan a la eficacia de las alternativas sostenibles.
Pheromone-mediated mating disruption (MD), a pest management tactic that prevents moth mating, has helped reduce insecticide use and crop damage in agriculture. The need to decrease the economic and labor costs associated with its application has encouraged the development of automated dispensers. These puffers disrupt mating behavior of moth pests by releasing pheromone during the time when insects are active. However, the lack of a firm basis on the biology and behavior of moths, and how external factors can alter their periodicity, prevent the proper adjustment of the puffers, and may result in a decrease of their efficacy in the field. Although automated traps can be used to determine these activity periods and adjust puffer pheromone release, commercially automated traps do not offer enough temporal resolution to outline insect activity periods, which usually last one or a few hours. In my thesis, we have designed and tested a cheap and easy to build high temporal resolution image-sensor insect trap. Based on captures in traps lured with synthetic pheromone and virgin females I have determined the daily and seasonal Grapholita molesta male sexual responsiveness. Although the lure did not influence the daily and seasonal sexual responsiveness period of Grapholita molesta (Busk), the flight activity within 24 h circadian rhythm is often influenced by sunset time and temperature. Most captures were registered between 3 hours before and 1 hour after sunset, indicating a diurnal to crepuscular activity of the species. Lure type (septum or female) did not influence the daily time of flight. While low temperatures led to an earlier flight in males, warmer temperatures resulted in flights closer to the sunset time. To reduce the cost of MD and avoid wasting pheromone, puffers should be programmed to spray at a variable time throughout the season, following the curve of activity of the insects. In order to explain the mechanisms of pheromone-mediated MD in pest management, most studies have focused on the effect high levels of synthetic pheromones have on the behavior of males, whereas females were considered unresponsive to their own pheromone. However, a recent review shows that there is substantial behavioral evidence that female moths respond to their own sex pheromone. Nevertheless, the evidence for sex pheromone "autodetection" at the olfactory receptor neuron (ORN) level is limited. By means of electrophysiological methods I compared the responses of ORNs housed in antennal sensilla trichodea to an array of biologically relevant compounds of male and female G. molesta, a species with reported pheromone autodetection. Hierarchical cluster analysis (HCA) indicated a radically different peripheric olfactory system between sexes that could be related to their specific ecological roles. In males no cells responded to their own courtship pheromone ethyl trans-cinnamate, while most (63%) were tuned specifically to the major or minor pheromone compounds (Z8-12:Ac and E8-12:Ac, respectively), their relative abundance being similar to their ratio in the female pheromone. Plant volatile cells were relatively frequent in females (6%) and 3% of the female ORNs were also tuned to the male-produced courtship pheromone. Several female cells were excited by female-produced sex pheromone, but their responses were generally lower than in males, and they responded broadly to the other compounds as well, so the HCA grouped them in a large cluster (91%) of "unspecific" female neurons. The lack of differential sex pheromone receptor neurons in females, and their relatively low sensitivity to sex pheromone would not allow females to detect variations in the pheromone ratios in nature. Thus, the alteration of their behavior after exposure to conspecific pheromone under laboratory conditions does not appear to be species-specific. Female behavior can nonetheless be specifically altered by other cues more relevant to their biological needs. The headspace of different plants may induce a different effect on females stemming from different hosts. I analyzed electroantennogram (EAG) responses of male and female adults of the European grapevine moth Lobesia botrana (Denis et Schiffermüller) (Lepidoptera: Tortricidae) collected as larvae from grapevine (Vitis vinifera L.) and flax-leaved daphne (Daphne gnidium L.) to specific and shared plant volatiles of the two hosts, as well as to sex pheromone compounds. My results indicate that pheromone detection did not differ between the two populations. Furthermore, host-plant volatile detection was not affected by sex or larval host plant. Polyphagous populations developing on a specific host seem therefore able to detect the plant volatiles of alternative hosts. However, lack of statistical differences in odor discrimination at the antennal level does not imply that insects from each host would show similar preference for the two host-specific odor blends, since behavioral responses to plant odors require brain integration of the antennal input of each individual odorant in a blend. Differences in plant preference could still occur if there are no differences at the EAG level. Understanding the potential ability of a polyphagous moth to reproduce or find food and shelter in alternative hosts is important to assess the topographical limitations of MD. My thesis serves to understand a bit better the complex functioning of a technique like MD and the repercussions that some environmental factors can have in its proper functioning. Even though pesticide use has been reduced, we still rely on these chemicals to prevent pest outburst. A more deep and rigorous study of the variables affecting the efficacy environmentally friendly alternatives should be conducted.

The effects of pheromone trap placement on male moth catches of both species were studied. For P. flavedana, traps hung at 2.1 and 3.0 meters captured the greatest number of moths. Trap heights of 1.2, 2.1, and 3.0 meters caught the greatest number of P. idaeusalis moths. The outside-the-canopy trap position captured more P. flavedana moths, while the within-canopy trap location caught the greatest number of P. idaeusalis moths. Traps placed in the west portion of the tree captured the greatest number of P. flavedana moths. P. idaeusalis moth catches were not influenced by compass quadrants. Trap design and pheromone dispenser and rate influenced trap catches of P. flavedana. Development of P. flavedana and P. idaeusalis on a meridic diet was observed at constant temperatures in the laboratory. Lower developmental threshold values for egg, larval, and pupal stages of P. flavedana were: 10.6, 8.6, 9.0°C, respectively. Lower developmental threshold values of 9.7, 7.0, and 8.5°C were estimated for P. idaeusalis egg, larval, and pupal stages, respectively. An average of 101.5 °D_10.6’, 379.6 °D_8.6’, and 126.0 °D_9.0’ were required for development of egg, larval, and pupal stages of P. flavedana, respectively. P. idaeusalis required 104.7 °D_9.7’, 442.7 °D_7.0’, and 132.2 °D_8.5’ to complete development in the egg, larval, and pupal stages, respectively. Differences in rate of development were observed between food sources for both species. Within-tree spatial distribution of egg masses and fruit damage resulting from larval feeding for both species was investigated. P. flavedana egg masses were mostly found in the southern portion of the tree below 1.8 meters. Egg masses of P. idaeusalis were observed in greatest numbers in the southern and eastern quadrants of the tree below 2.8 meters. Fruit damage caused by larvae of both species was greatest in the lower portion of the tree. Wind dispersal of first-instar larvae between trees is believed to have influenced fruit damage distribution. The seasonal activity of P. flavedana and P. idaeusalis was monitored. Degree-day accumulations for first moth catch, first and peak egg deposition, and first and peak egg hatch of both generations are presented.
Ph. D.

The biology of key citrus pests Thaumatotibia leucotreta Meyrick (Lepidoptera: Tortricidae), Ceratitis capitata Wiedemann (Diptera: Tephritidae) and Ceratitis rosa Karsch (Diptera: Tephritidae) includes their dropping from host plants to pupate in the soil below citrus trees. Since most EP fungi are soil-borne microorganisms, the development and formulation of alternative control strategies using these fungi as subterranean control agents, targeted at larvae and pupae in the soil, can potentially benefit existing IPM management of citrus in South Africa. Thus, a survey of occurrence of entomopathogenic fungi was undertaken on soils from citrus orchards and natural vegetation (refugia) on conventionally and organically managed farms in the Eastern Cape Province in South Africa. A method for baiting soil samples with citrus pest T. leucotreta and C. capitata larvae, as well as with the standard bait insect, Galleria mellonella Linnaeus (Lepidoptera: Pyralidae), was implemented. Sixty-two potentially useful entomopathogenic fungal isolates belonging to four genera were collected from 288 soil samples, an occurrence frequency of 21.53%. The most frequently isolated entomopathogenic fungal species was Beauveria bassiana (Balsamo) Vuillemin (15.63%), followed by Metarhizium anisopliae var. anisopliae (Metschnikoff) Sorokin (3.82%). Galleria mellonella was the most effective insect used to isolate fungal species (χ2=40.13, df=2, P≤ 0.005), with a total of 45 isolates obtained, followed by C. capitata with 11 isolates, and T. leucotreta with six isolates recovered. There was a significantly (χ2=11.65, df=1, P≤ 0.005) higher occurrence of entomopathogenic fungi in soil samples taken from refugia compared to cultivated orchards of both organically and conventionally managed farms. No significant differences were observed in the recovery of fungal isolates when soil samples from both farming systems were compared. The physiological effects and host range of 21 indigenous fungal isolates obtained in the Eastern Cape were investigated in the laboratory to establish whether these isolates could be effectively used as biological control agents against the subterranean life stages of C. rosa, C. capitata and T. leucotreta. When these pests were treated with a fungal concentration of 1 x 10⁷ conidia ml⁻¹, the percentage of T. leucotreta adults which emerged in fungal treated sand ranged from 5 to 60% (F=33.295; df=21; P=0.0001) depending on fungal isolate and the percentage of pupae with visible signs of mycosis ranged from 21 to 93% (F= 96.436; df=21; P=0.0001). Based on fungal isolates, the percentage adult survival in C. rosa and C. capitata ranged from 30 to 90% and 55 to 86% respectively. The percentage of C. rosa and C. capitata puparia with visible signs of mycosis ranged from 1 to 14% and 1 to 11% respectively. Deferred mortality due to mycosis in C. rosa and C. capitata adult flies ranged from 1 to 58% and 1 to 33% respectively, depending on fungal isolate. Entomopathogenic fungal isolates had a significantly greater effect on the adults of C. rosa and C. capitata than they did on the puparia of these two fruit fly species. Further, C. rosa and C. capitata did not differ significantly in their response to entomopathogenic fungi when adult survival or adult and pupal mycosis were considered. The relative potency of the four most virulent Beauveria isolates as well as the commercially available Beauveria bassiana product, Bb Plus® (Biological Control Products, South Africa), were compared against one another as log-probit regressions of mortality against C. rosa, C. capitata and T. leucotreta which all exhibited a dose-dependent response. Against fruit flies the estimated LC50 values of all five Beauveria isolates ranged from 5.5 x 10¹¹ to 2.8 x 10¹² conidia/ml⁻¹. There were no significant differences between the relative potencies of these five fungal isolates. When T. leucotreta was considered, isolates: G Moss R10 and G 14 2 B5 and Bb Plus® were significantly more pathogenic than G B Ar 23 B3 and FCM 10 13 L1. The estimated LC₅₀ values of the three most pathogenic isolates ranged from 6.8 x 10⁵ to 2.1 x 10⁶ conidia/ml⁻¹, while those of the least pathogenic ranged from 1.6 x 10⁷ to 3.7 x 10⁷ conidia/ml⁻¹. Thaumatotibia leucotreta final instar larvae were exposed to two conidial concentrations, at four different exposure times (12, 48, 72 and 96 hrs) and showed an exposure time-dependant relationship (F=5.43; df=3; P=0.001). At 1 x 10⁷conidia/ml⁻¹ two Beauveria isolates: G Moss R10 and G 14 2 B5 were able to elicit a response in 50% of test insects at 72 hrs (3 days) exposure. Although a limited amount of mycosis was observed in the puparia of both fruit fly species, deferred adult mortality due to mycosis was high. The increased incidence of adult mortality suggests that post emergence mycosis in adult fruit flies may play a more significant role in field suppression than the control of fruit flies at the pupal stage. The increased incidence of pupal mortality, as well as the relatively low concentrations of conidia required to elicit meaningful responses in T. leucotreta pupae may suggest that pre-emergent control of false codling moth will play a more significant role in field suppression than the control of adult life stages using indigenous isolates of entomopathogenic fungi. Various entomopathogenic fungal application techniques targeted at key insect pests within integrated pest management (IPM) systems of citrus are discussed.

Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is one of the major citrus pests of economic importance for South Africa’s citrus industry. It is endemic to Africa, and therefore a phytosanitary pest with zero tolerance by most export markets. The cryptic nature of T. leucotreta makes visual inspection an inefficient method for detecting neonate larvae in fruit in the packhouse. Therefore, a more accurate method for sorting infested fruit at the packhouse, particularly for newly infested fruit could ensure market access. A recent study showed that fruit infested by T. leucotreta emit a chemical profile different from that of a healthy fruit. Several studies provide evidence that parasitoids locate their hosts feeding on fruit by exploiting the novel chemical profiles produced due to host herbivory. The aim of this study was to evaluate the potential of using the naturally occurring behaviour of a larval parasitoid Agathis bishopi (Nixon) (Hymenoptera: Braconidae) for detection of T. leucotreta infested fruit, by determining which compound in infested fruit is attractive to parasitoids. Ytube olfactometer and flight-tunnel bioassays with healthy and T. leucotreta infested fruit showed a significantly stronger response of A. bishopi female parasitoids to infested fruit. Among the volatile compounds associated with T. leucotreta infested fruit, D-limonene elicited the strongest attraction to A. bishopi female parasitoids. Attraction of mated A. bishopi female parasitoids to T. leucotreta infested fruit and D-limonene significantly increased after oviposition experience. Behavioural responses of A. bishopi female parasitoids that were associated with T. leucotreta infested fruit were investigated to determine which behaviours are distinct and interpretable. Probing and oviposition behaviours were the most noticeable and were only elicited on infested fruit when parasitoids contacted T. leucotreta frass, indicating that chemical compounds in frass are short-range cues used for final host location. Since production of D-limonene by fruit is elevated due to herbivory by different pests including mechanical injury on fruit, response of A. bishopi female parasitoids to compounds in frass offers a more specific and potentially useful mechanism for development of a detection system for T. leucotreta infested fruit. Chemical analysis of T. leucotreta frass and conditioning A. bishopi parasitoids to respond behaviourally to compounds in frass is proposed.

False codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is considered the most important indigenous pest of citrus in southern Africa. Major concerns such as progressive insecticidal resistance, the negative impact of insecticides on the environment, as well as the influence of consumers opposed to chemical residues on fruit, created opportunities for biological control methods such as Sterile Insect Technology (SIT). This technology is now established in the Western and Eastern Cape provinces of South Africa as an effective, sustainable alternative to conventional FCM control methods. Due to the prevalence of the pest in all citrus producing areas of South Africa, potential for SIT to expand is enormous. Success of an SIT programme is highly dependent on efficient application of the technology to achieve its objectives in a timeous manner. The aim of this study was to advance the application of SIT for control of FCM on citrus in South Africa, by investigating the effect of certain critical stages in the process. The effect of long-distance transportation on fitness of irradiated FCM was determined, showing reduced performance with cold-immobilized transport. A significant decrease in flight ability and longevity of irradiated FCM was found, although critically, realized fecundity was not affected. The effect of two different insecticides in the pyrethroid and organophosphate chemical groups were investigated for their residual effect on mortality of released irradiated FCM, to determine if these pest control programmes could be integrated. Both chlorpyrifos and tau-fluvalinate were effective in killing irradiated FCM for a number of days after application, after which degradation of the active ingredient rendered it harmless. This effect was found to be similar for irradiated and non-irradiated males, consequently ratios of sterile : wild male FCM should be retained regardless of whether sprays are applied or not. The modes for release of sterile FCM in an SIT programme were investigated. Efficacy of ground and aerial release platforms were tested by evaluating the recovery of released irradiated male FCM in these orchards. More irradiated FCM were recovered in orchards released from the ground compared to air. However, an economic analysis of both methods shows application of irradiated insects over a large geographical area is more cost-effective by air. Depending on the terrain and size of the target area, a combination of both methods is ideal for application of SIT for control of FCM in citrus. Development of application technology for advance of the programme is discussed and recommendations for future research and development are offered.

Dissertation (PhD(Agric))--Stellenbosch University, 2003.
ENGLISH ABSTRACT: The codling moth, Cydia pomonella (L.) has been a major pest of pome fruits since before the turn of the last century. However, despite its high economic profile little is known about the bionomics and . behaviour of this pest in apple orchards in South Africa, information required for the development of a sustainable integrated management programme. In field trials there was contingency between the time of year and the upper and lower half of the tree. First generation moths laid significantly more eggs in the bottom half of the tree while second and third generation moths laid significantly more eggs in the top half of the tree. The preferred oviposition sites on Granny Smith (GS) and Golden Delicious (GD) cultivars, in order of preference, were leaves, fruit and wood. More eggs were laid on the fruit ofGS spurs (35.6 %) than on those ofGD spurs (10.7 %). On fruit spurs there was a significant increase in the number of eggs on GD leaves and GS fruit over the season, whereas the number of eggs on GS leaves and GD fruit remained constant. On branches there was an increase in the number of eggs on GD and GS leaves, but not on the fruit or wood. The preferred oviposition site on the fruit was the fovea of the stalk insertion and the rounded cheek area surrounding the fovea. The distribution within different fruit bearing classes (1 - 4 fruit per spur) was random only for one fruit per spur, while on the other spur classes clustering occurred. In laboratory studies of the embryonic and immature stages there was a linear relationship between rate of development and constant temperatures of 15, 17,20,25 and 30·C ± l·C. The lower threshold temperatures for embryonic, larval and pupal development were 11.1, 7.9, 9.9°C respectively. The degree-days required to complete embryonic, larval and pupal development were 80.5, 345, and 279 respectively. The response of the different stages to constant temperatures was similar to that under fluctuating temperatures. At temperatures below 16°C or abouve 27°C moths did not mate and few eggs were laid. Moth longevity decreased with increasing temperature. There was seasonal variation in longevity and oviposition at constant and fluctuating temperatures. Summer adults produced significantly more eggs than spring adults at constant temperatures. At a constant temperature of2S·C and fluctuating temperatures there were five distinct larval instars. The similarity between the mean head capsule width and ranges for each instar reared on fruit of different stages of development at fluctuating temperatures indicates that fruit development and temperature have little influence on mean head capsule width. From sleeve-cage studies in the orchard there was no significant difference in the fecundity of spring and summer moths. In the beginning of October spring moths produced significantly fewer eggs than in November. Egg mortality increased from 8.2 %in spring to 21.2 %in summer. Failure of 1st instar larvae to penetrate the fruit ranged from 4.9 % to 19.5 %, while mortality oflarvae from egg hatch to emergence from the fruit ranged from 29.7 % to 42.9 %. Mortality of 5th instar larvae after emerging from the apples ranged from 0 % to 8.7 % and pupal mortality from 0 % to 3.5 %. On large 27-year old trees more overwintering larvaewere found on Golden Delicious (13.9) than on Granny Smith trees (5.7), with over 70 % oflarvae being found in pruning wounds on both cultivars. On small 7-year old Golden Delicious and Granny Smith trees the mean number oflarvae was 0.5 and 2.0 per tree. A combined mating disruption and insecticide control programme reduced codling moth resistant populations to levels requiring a minimum to no insecticide intervention for several seasons. The efficacy of a pheromone based strategy, number of pheromone treatments, number of dispenserslha and level of insecticide intervention required are strongly influenced by prevailing weather conditions. Fruit infestation in orchards under a mating disruption programme and under an insecticide programme were greater along the borders compared to the interior. The presence of horticultural mineral oil on the leaves and branches did not have a detrimental effect on oviposition nor was there any significant ovicidal effect. A significant ovicidal effect was obtained when applied after oviposition. In field trials, insecticides with lower levels of efficacy than the primary insecticide, azinphos-methyl, provided acceptable control when successfully incorporated into a spray programme which followed a policy of alternation of insecticides across generations. The least variation in the number of degree-days between biofix and first egg hatch of the spring flight was when the second trap catch (Biofix 2) was used as the biofix. A biofix based on the first evening when the temperature reached or exceeded 1TC at 18:00 after first trap catch also showed less variation than when the biofix was based on first trap catch. The mean number of degree-days accumulated between Biofix 2 and first egg hatch was found to be 139.1 ° D. The number of degreedays between the first and second flight biofixes varied between 531.2 and 488.87°D with a mean of 508.1°D.
AFRIKAANSE OPSOMMING: Kodlingmot (Cydia pomonella) is reeds sedert die vorige eeu 'n sleutelplaag van kemvrugte. Ten spyte van die hoë ekonomiese profiel, is daar min kennis betreffende die bionomie en gedrag van hierdie plaag in appelboorde in Suid-Afrika. Hierdie inligting is noodsaaklik vir die ontwikkeling van 'n volhoubare geïntegreerde bestuursprogram. Tydens veldproewe is 'n korrelasie tussen die tyd van die jaar en eierlegging in die boonste of onderste helfte van bome gevind. Eerste-generasie motte het betekenisvol meer eiers in die onderste helfte van die bome gelê, terwyl tweede- en derde-generasie motte meer eiers in die boonste helfte van die bome gelê het. In toenemende volgorde was die voorkeur eierleggingsposisies op Granny Smith (GS) en Golden Delicious (GD) appels die blare, vrugte en hout. Meer eiers is op vrugte van GS spore (35.6%) as op dié van GD spore (10.7%) gelê. Op vrugspore was daar 'n betekenisvolle toename in aantal eiers op GD blare en GS vrugte deur die seisoen, terwyl die getal eiers op GS blare en GD vrugte konstant gebly het. Op takke was daar 'n toename in aantal eiers op GD en GS blare, maar nie op vrugte of hout nie. Die voorkeur eierleggingsposisie op vrugte was die fovea van die steelaanhegting en die wang rondom die fovea. Die verspreiding tussen verskillende vrugdraende klasse (1 - 4 vrugte per spoor) was slegs in die een-Vrug-per-spoorklas ewekansig, terwyl daar in die ander spoorklasse groepering voorgekom het. In laboratoriumstudies van embrioniese en onvolwasse stadia is 'n lineêre verband tussen ontwikkelingskoers en konstante temperature van 15, 17,20,25 en 30°C±I°C gevind. Die onderste drempel-temperature vir embrioniese, larwale en papie-ontwikkeling was onderskeidelik II. 1°C, 7.f?C en 9.9°C. Graaddae benodig vir voltooiing van embrioniese, larwale en papie-ontwikkeling was onderskeidelik 80.5,345 en 279. Die respons van verskillende stadia by konstante temperature het ooreengestem met hul respons onder wisselende temperature. Motte het nie gepaar nie en min eiers is gelê by temperature onder 16°C of bo 27°C. Die lewensverwagting van motte het afgeneem met toename in temperatuur. Seisoenale variasie in . lewensverwagting en eierlegging het voorgekom by konstante sowel as wisselende temperature. By konstante temperature het somer-volwassenes betekenisvol meer eiers as lente-volwassenes geproduseer. By 'n konstante temperatuur van 25°C, sowel as by wisselende temperature, het vyf duidelik onderskeibare larwale instars voorgekom. Die ooreenkoms tussen die gemiddelde kopkapsulewydte en wydte-reeks vir elke instar wat op vrugte van verskillende stadiums van ontwikkeling by wisselende temperature geteel is, dui daarop dat vrugontwikkeling en temperatuur weinig invloed op gemiddelde kopkapsule-wydte het. Tydens mou-hok studies in die boord is geen betekenisvolle verskil in die fekunditeit van lente- en somer-motte waargeneem nie. Vroeg in Oktober het lente-motte betekenisvol meer eiers as in November geproduseer. Eiermortaliteit het van 8.2% in die lente tot 21.2% in die somer toegeneem. Faling van 1ste instar larwes om vrugte te penetreer het van 4.9% tot 19.5% gewissel, terwyl mortaliteit van larwes vanaf uitbroei tot uitkoms uit die vrug van 29.7% tot 42.9% gewissel het. Mortaliteit van Sde instar larwes na uitkoms uit die vrug het van 0% tot 8.7% gewissel, en papie-mortaliteit van 0% tot 3.5%. Op groot, 27-jaar oue bome is meer oorwinterende larwes op Golden Delicious (13.9) as op Granny Smith (5.7) gevind, en meer as 70% van die larwes op beide kultivars is op snoeiwonde gevind. Op klein, 7-jaar oue bome was die gemiddelde aantallarwes op Golden Delicious en Granny Smithbome 0.5 en 2.0 onderskeidelik. 'n Gekombineerde paringsontwrigting- en insekdoder beheerprogram het weerstandbiedende kodlingmot-populasies verminder tot 'n vlak waar minimum tot geen insekdoder-toedienings vir verskeie seisoene gemaak is. Die effektiwiteit van 'n feromoon-gebaseerde strategie, aantal feromoonbehandelings, aantal vrystellers/ha en vlak van insekdoder-toediening word sterk deur heersende weersomstandighede beïnvloed. Die rande het hoër vruginfestasie as die middel getoon in boorde onder paringsontwrigting sowel as boorde onder insekdoder-programme. Die teenwoordigheid van minerale olie op blare en takke het geen nadelige effek op eierlegging gehad nie en dit het geen betekenisvolle eierdodende effek gehad nie. Indien die olie ná eierlegging toegedien is, is daar wel 'n betekenisvolle eierdodende effek waargeneem. Tydens veldproewe het insekdoders met laer effektiwiteit as die primêre insekdoder, azinfos-metiel, aanvaarbare beheer verskaf indien dit suksesvol geïnkorporeer is in 'n spuitprogram deur 'n beleid van afwisseling van insekdoders oor generasies. Die kleinste variasie tussen die aantal graaddae tussen biofix en eerste uitbroei van eiers is gevind indien die tweede lokvalvangs as biofix gebruik is. 'n Biofix gebaseer op die eerste aand na die eerste lokval vangste wat die temperatuur 17°Cofhoër was teen 18:00, het ook 'n kleiner variasie getoon as die eerste lokvalvangs. Die aantal graaddae tussen die tweede en derde vlug biofix het tussen 531.2 en 488.87°D gewissel, met 'n gemiddelde van 508.1°D.

Com o objetivo de avaliar o potencial de utilização de espécies/linhagens de Trichogramma como mais um componente do manejo de Ecdytolopha aurantiana (Lima, 1927), importante praga da citricultura paulista, foram realizadas avaliações biológicas envolvendo seleção de linhagens, biologia em diferentes temperaturas, exigências térmicas, capacidade de parasitismo e número de ideal de parasitóides a ser liberado por ovo da praga. A seleção de linhagens/espécies de Trichogramma é básica para o controle de E. aurantiana, pois ocorreu uma grande variação no parasitismo de ovos da praga dentre os vários materiais biológicos estudados. A razão sexual das espécies/linhagens não foi afetada pela temperatura na faixa de 18 a 32ºC, sendo a temperatura de 25ºC a mais adequada para as espécies/linhagens estudadas. A duração do período ovo-adulto de T. atopovirilia Oatman & Platner, 1983 e de T. pretiosum Riley, 1879 foi inversamente relacionada como aumento da temperatura, na faixa térmica de 18 a 32ºC, sendo as exigências térmicas dessas espécies muito próximas, em torno de 108 GD. O hospedeiro de criação natural, E. aurantiana ou o alternativo, Anagasta kuehniella (Zeller, 1879) não afetaram o parasitismo das espécies/linhagens selecionadas. Em laboratório, obteve-se maior parasitismo expondo-se 30 ovos de E. aurantiana por fêmea de Trichogramma spp. durante 34 horas. As maiores taxas de parasitismo de ovos de bicho-furão para as duas espécies/linhagens ocorreram quando os parasitóides foram provenientes de ovos de E. aurantiana. A proporção estimada de 36 parasitóides por ovo de E. aurantiana possibilitou a maior porcentagem de parasitismo por T. pretiosum. Os resultados obtidos indicaram que os parasitóides de ovos do gênero Trichogramma apresentam potencial de controle de E. aurantiana, desde que liberados em grandes quantidades por unidade de área e em torno das plantas.
In order to evaluate the potential of use of Trichogramma species/strains as an additional component in the management of Ecdytolopha aurantiana (Lima, 1927), an important pest of Citrus in the state of São Paulo, biological trials were carried out involving strain selection, biology at different temperatures, thermal requirements, parasitism capacity and optimal number of parasitoids to be released per pest egg. The selection of Trichogramma strains/species is basic to control E. aurantiana for the egg parasitism of the pest varied greatly among several biological materials studied. The sex ratio of the species/strains was not affected by the temperature at a 18-32ºC range, and 25ºC was the more suitable temperature for the species/strains studied. The duration of the egg-adult period of T. atopovirilia Oatman & Platner, 1983 and T. pretiosum Riley, 1879 was inversely related to the increased temperature at a thermal range of 18-32ºC, and the thermal requirements of these species are very close, ranging 108 DD (degree days). Neither the natural rearing host, E. aurantiana, nor the factitious host, Anagasta kuehniella (Zeller, 1879) affected the parasitism of the selected species/strains. The proportion of 30 eggs of E. aurantiana per Trichogramma spp. female provided a higher parasistism rate, with eggs exposed to the parasitoid for 34 hours. The higher parasitism rates of the eggs of citrus fruit borer for the two species/strains occurred when the parasitoids were reared on eggs of E. aurantiana. The estimated ratio of 36 parasitoids per egg of E. aurantiana allowed a higher parasitism percentage of T. pretiosum. The results achieved showed that the egg parasitoids of Trichogramma have a potential to control E. aurantiana provided that they are released in large amounts per area unit and surrounding the plants.

A lagarta-enroladeira Bonagota salubricola (Meyrick, 1937) e a mariposa oriental Grapholita molesta (Busck, 1916) (Lepidoptera: Tortricidae) são duas das mais importantes pragas de frutíferas de clima temperado no Brasil. O manejo destes insetos-praga tem sido realizado quase que exclusivamente com inseticidas. Para a implementação de estratégias de manejo pró-ativo de resistência, é importante conhecer o estado atual de suscetibilidade destas espécies a inseticidas para detectar a resistência antes que se observem falhas no controle. Nas condições brasileiras, em regiões onde estes pomares são plantados próximos em algumas regiões, acredita-se que G. molesta disperse para a cultura da maçã após a colheita do pêssego, dependendo da proximidade espacial e temporal dos pomares. Portanto, este trabalho teve como objetivos, caracterizar a suscetibilidade de B. salubricola e G. molesta aos principais inseticidas recomendados para o controle e avaliar a estrutura genética de populações de G. molesta provenientes das culturas da macieira e pessegueiro no Brasil. A caracterização da suscetibilidade foi realizada com bioensaio de ingestão com tratamento superficial da dieta com inseticidas, utilizando-se lagartas neonatas provenientes de populações de B. salubricola coletadas em macieira no Estado do Rio Grande do Sul (safra 2011/12) e de G. molesta provenientes de pomares do Rio Grande do Sul (2010/11 e 2011/12), Santa Catarina (2010/11) e São Paulo (2010/11 e 2011/12). Não foram observadas diferenças na suscetibilidade de populações de campo de B. salubricola em relação à população suscetível de referência aos inseticidas chlorantraniliprole, phosmet, spinetoram, spinosad e tebufenozide, assim como não foram detectadas diferenças na suscetibilidade de populações de G. molesta a chlorantraniliprole, metaflumizone, novaluron, pyriproxyfen e spinetoram. Entretanto, foram verificadas diferenças significativas na sobrevivência de populações de B. salubricola ao novaluron (3,3% de sobrevivência) e de G. molesta aos inseticidas phosmet e tebufenozide, com 2,5 e 4,5% de sobrevivência, respectivamente. Portanto, a frequência de resistência de B. salubricola e G. molesta ainda é baixa aos inseticidas avaliados. Com base nos marcadores mitocondriais e microssatélites foi possível detectar estruturação genética significativa entre as populações de G. molesta que infestam as culturas da macieira e pessegueiro, com indicação de estruturação em função de hospedeiros (?ST = 0,198; P < 0,05) e da distância geográfica (r=0,545; valor de p<0,001). Em geral, a variabilidade genética de G. molesta foi bem distribuída nas regiões produtoras de maçã e pêssego e as barreiras geográficas, as condições edafoclimáticas e o manejo da praga parecem estar limitando o acasalamento entre os indivíduos das populações distintas avaliadas. Portanto, devido ao baixo fluxo gênico entre as populações de G. molesta, as estratégias de manejo da resistência podem ser implementadas no âmbito local para essa praga.
The apple leafroller Bonagota salubricola (Meyrick, 1937) and Oriental fruit moth Grapholita molesta (Busck, 1916) are two of the most important pests of temperate fruit trees in Brazil. Management of these both insect pest has been conducted almost exclusively with insecticides. For the implementation of proactive resistance management strategies, it is important to know the current status of pest susceptibility to insecticides to detect the resistance before control failures with the use of insecticides. In Brazilian conditions, where apple and peach orchards are very often planted close in some regions, it is believed that G. molesta may disperse to apple orchards after harvesting peaches, depending on the spatial and temporal proximity of orchards. Therefore, the objectives of this study were to characterize the susceptibility of B. salubricola and G. molesta to the main insecticides recommended for their control and to evaluate the genetic structure of G. molesta populations from the apple and peach orchards in Brazil. B. salubricola populations were collected in apple orchards in the State of Rio Grande do Sul (2011/12 growing season) and G. molesta populations from orchards in Rio Grande do Sul (2010/11 and 2011/12), Santa Catarina (2010/11) and São Paulo (2010/11 and 2011/12). There were no differences in the susceptibility among field populations of B. salubricola in compared to the susceptible reference population to the insecticides chlorantraniliprole, phosmet, spinetoram, spinosad and tebufenozide and among G. molesta populations to chlorantraniliprole, metaflumizone, pyriproxyfen and spinetoram. However, there were significant differences in survival of B. salubricola populations to novaluron (3.3% survival) and G. molesta populations to insecticides phosmet and tebufenozide, with 2.5 and 4.5% survival, respectively. Therefore, the frequency of resistance of B. salubricola and G. molesta is still low to insecticides evaluated herein. Based on mitochondrial and microsatellites markers, significant genetic structure among G. molesta populations was detected based on the host plant (?ST = 0,198; P < 0,05) and the geographic distance (r=0,545; valor de p<0,001). In general, the genetic variability of G. molesta is well distributed in the producing regions of apple and peach and the geographic barriers, soil and climatic conditions and pest management can be limiting the mating among individuals from distinct populations evaluated in this study. Therefore, due to low gene flow among G. molesta populations in Brazil, resistance management strategies can be implemented at local level.

The control of false codling moth (FCM), Thaumatotibia leucotreta (Meyrick, 1912) (Lepidoptera: Tortricidae), in citrus orchards is strongly reliant on the use of integrated pest management as key export markets impose stringent chemical restrictions on exported fruit and have a strict no entry policy towards this phytosanitary pest. Most current, registered control methods target the above-ground life stages of FCM, not the soil-dwelling life stages. As such, entomopathogenic fungi which are ubiquitous, percutaneously infective soil-borne microbes that have been used successfully as control agents worldwide, present ideal candidates as additional control agents. Following an initial identification of 62 fungal entomopathogens isolated from soil collected from citrus orchards in the Eastern Cape Province, South Africa, further laboratory research has highlighted three isolates as having the greatest control potential against FCM subterranean life stages: Metarhizium anisopliae G 11 3 L6 (Ma1), M. anisopliae FCM Ar 23 B3 (Ma2) and Beauveria bassiana G Ar 17 B3 (Bb1). These isolates are capable of causing above 80% laboratory-induced mycosis of FCM fifth instars. Whether this level of efficacy was obtainable under sub-optimal and fluctuating field conditions was unknown. Thus, this thesis aimed to address the following issues with regards to the three most laboratory-virulent fungal isolates: field efficacy, field persistence, optimal application rate, application timing, environmental dependency, compatibility with fungicides and the use of different wetting agents to promote field efficacy. Following fungal application to one hectare treatment blocks in the field, FCM infestation within fruit was reduced by 28.3% to 81.7%. Isolate Bb1 performed best under moderate to high soil moisture whilst Ma2 was more effective under low soil moisture conditions. All isolates, with the exception of Ma2 at one site, were recorded in the soil five months post-application. None of the wetting agents tested were found to be highly toxic to fungal germination and similar physical suspension characteristics were observed. Fungicide toxicity varied amongst isolates and test conditions. However, only Dithane (a.i. mancozeb) was considered incompatible with isolate Ma2. The implication of these results and the way forward is discussed. This study is the first report of the field efficacy of three laboratory-virulent fungal isolates applied to the soil of conventional citrus orchards against FCM soil-dwelling life stages. As such, it provides a foundation on which future research can build to ensure the development and commercialisation of a cost-effective and consistently reliable product.

La carpocapsa, Cydia pomonella (L.) (Lepidoptera: Tortricidae), és una de les plagues més importants del cultiu dels fruiters de pinyol, principalment de la pomera. A la zona fructícola de Lleida, a l’igual que a la majoria de les zones productives del món, s’han detectat problemes de resistència a insecticides piretroïds, organofosforats, benzoilurees, fenoxycarb i diacilhidracines. El principal mecanisme de resistència detectat ha estat l’acció dels enzims detoxificadors multifunció-oxidases (MFO) i, de manera menys important, glutathion transferases (GST) i esterases (EST). A la present tesis doctoral, s’han determinat els nivells resistència de carpocapsa a insecticides de baix risc per a la salut humana i medi ambient i la presència i extensió de les mutacions de la acetilcolinesterasa (AChE) i la knockdown (kdr), en poblacions de camp procedents de tres zones productives de poma a España, amb sistemes de maneig molt diferents. Les poblacions asturianes resultaren susceptibles a tots els insecticides provats, mentre que les poblacions de la Val de l’Ebre (Catalunya i Aragón) presentaren resistència en alguna població i tolerància de forma freqüent a metoxifenocide, lambda-cyhalotrin i tiacloprid. La resta de productes provats, etil-clorpirifos, tebufenocide, spinosad, spinetoram, indoxacarb, clorantraniliprole i emamectine, resultaren efectius fins i tot en les poblacions amb elevats nivells de resistència. La mutació AChE està present de forma generalitzada únicament a les parcel·les de la Val de l’Ebre, mentre que la kdr està present a la majoria de les parcel·les de Catalunya. A les poblacions d’ Asturias i a la població d’Extremadura analitzades no es detectaren mutacions. També s’ha determinat la línia base de susceptibilitat insecticida de la matèria activa chlorantraniliprole en poblacions de camp europees de carpocapsa, i s’ha avaluat l’evolució de la seva eficàcia al llarg dels anys d’aplicació. No s’han detectat poblacions resistents a chlorantraniliprole, però, tanmateix, sí s’ha detectat una relació entre la freqüència d’individus MFO-resistents (amb un elevat nivell d’enzims de detoxificació MFO) amb el percentatge de la mortalitat produïda per la concentració discriminant pròxima a la CL50. A més, s’estudià la freqüència d’adults MFO-resistents de C. pomonella capturats en trampes de feromones amb diferents difusors, amb i sense la cairomona (l’èster de pera, (2E, 4Z)-decadienoat de etil), degut a la major atracció d’aquests individus per la cairomona pura al laboratori. La finalitat fou la millora del monitoreig a les parcel·les amb una elevada presència d’individus resistents. Tanmateix, aquella major atracció no es produí al camp. Sí que es detectà una diferent freqüència d’individus resistents entre generacions i entre parcel·les amb o sense tractaments químics.
La carpocapsa, Cydia pomonella (L.) (Lepidoptera: Tortricidae), es una de las plagas más importantes del cultivo de los frutales de pepita, principalmente del manzano. En la zona frutícola de Lleida, al igual que en la mayoría de las zonas productivas del mundo, se han detectado problemas de resistencia a insecticidas piretroides, organofosforados, benzoilureas, fenoxycarb y diacilhidracinas. El principal mecanismo de resistencia detectado ha sido la acción de las enzimas detoxificadoras multifunción-oxidasas (MFO) y, en menor medida, glutathion transferasas (GST) y esterasas (EST). En la presente tesis doctoral, se han determinado los niveles de resistencia de carpocapsa a insecticidas de bajo riesgo para la salud humana y el medio ambiente y la presencia y extensión de las mutaciones de la acetilcolinesterasa (AChE) y la knockdown (kdr), en poblaciones de campo procedentes de tres zonas productivas de manzana en España, con sistemas de manejo muy distintos. Las poblaciones asturianas resultaron susceptibles a todos los insecticidas ensayados, mientras que las poblaciones del Valle del Ebro (Catalunya y Aragón) presentaron resistencia en alguna población y tolerancia de forma frecuente a metoxifenocide, lambda-cyhalotrin y tiacloprid. El resto de productos ensayados, etil-clorpirifos, tebufenocide, spinosad, spinetoram, indoxacarb, clorantraniliprole y emamectine, resultaron efectivos incluso en las poblaciones con elevados niveles de resistencia. La mutación AChE está presente de forma generalizada únicamente en las fincas del Valle del Ebro, mientras que la kdr está presente en la mayoría de las fincas en Catalunya. En las poblaciones de Asturias y la población de Extremadura analizadas no se detectaron mutaciones. También se ha determinado la línea base de susceptibilidad insecticida de la materia activa chlorantraniliprole en poblaciones de campo europeas de carpocapsa, y se ha evaluado la evolución su eficacia a lo largo de los años de aplicación. No se han detectado poblaciones resistentes a chlorantraniliprole, pero, sin embargo, sí se ha detectado una relación entre la frecuencia de individuos MFO-resistentes (con un elevado nivel de enzimas de detoxificación MFO) con el porcentaje de mortalidad de la concentración discriminante próxima a la CL50. Por otro lado, se estudió la frecuencia de adultos MFO-resistentes de C. pomonella capturados en trampas con distintos difusores, con y sin cairomona (el éster de pera, (2E, 4Z)-decadienoato de etilo), debido a una mayor atracción de estos individuos por la cairomona pura en laboratorio. La finalidad era la mejora del monitoreo en las fincas con una elevada presencia de insectos resistentes. Sin embargo, esa mayor atracción no se produjo en campo. Sí se detectó una distinta frecuencia de individuos resistentes entre generaciones y entre fincas con o sin tratamientos químicos.
Codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), is one of the most important pests of pome orchards, mainly of apple ones. Problems of resistance to insecticides pirethroids, organophosphates, benzoylureas, fenoxycarb, and diacylhydrazines have been detected in the fruit growing area of Lleida, as well as in the majority of the world apple growing areas. The main resistance mechanism detected has been the action of the detoxifying enzyms mixed-function oxidases (MFO) and, in a lesser extent, glutatione S-transferases (GST) and esterases (EST). In the present thesis, the levels of codling moth resistance to safe insecticides and the presence and the extension of the mutations acetylcholinesterase (AChE) and the knockdown (kdr) have been determined in field populations from three fruit growing areas of Spain, with very different production systems. The populations from Asturias were susceptible to all the tested insecticides, while some populations from the Ebro Valley (Catalonia and Aragón) were resistant, and frequently tolerant to methoxyfenozide, lambda-cyhalothrin and thiacloprid. The rest of tested insecticides chlorpyrifos-ethyl, tebufenozide, spinosad, spinetoram, indoxacarb, chlorantraniliprole and emamectin, were effective, even against populations with high resistance levels. The AChE mutation is generally present only in orchards from the Ebro Valley, while the kdr is present in the majority of the orchards from Catalonia. No mutations were detected in the populations from Asturias and Extremadura. The susceptibility baseline of chlorantraniliprole has been determined in field populations from Europe, and the evolution of its efficacy along time have been evaluated. Resistant populations to chlorantraniliprole have not been detected, but a relationship between the frequency of MFO-resistant individuals (those that show a high level of MFO detoxification enzymes) and the mortality (percentage) caused the discriminant concentration near to the LC50. Furthermore, the frequency of C. pomonella MFO-resistant adults caught in traps with different lures, with and without the kairomone (pear ester, (2E, 4Z)-2,4-decadienoate) was studied, because a higher attraction of these individuals for the pure kairomone had been detected in the laboratory. The aim was to improve monitoring in the orchards that show a high percentage of resistant individuals. However, this higher attraction was not observed in the field. A different frequency of resistant individuals among generations and among orchards chemically or non-chemically treated was observed.

Baculoviruses are arthropod-specific DNA viruses that are highly virulent to most lepidopteran insects. Their host specificity and compatibility with IPM programmes has enabled their usage as safe microbial insecticides (biopesticides). Two baculovirus-based biopesticides, Cryptogran and Cryptex, which have been formulated with Cryptophlebia leucotreta granulovirus (CrleGV) have been registered for the control of false codling moth (FCM), Thaumatotibia (=Cryptophlebia) leucotreta (Meyrick) (Lepidoptera: Tortricidae) in South Africa and have been successfully incorporated into IPM programmes. However, several studies have indicated that insects can develop resistance to baculovirus-based biopesticide as was shown with field populations of codling moth (CM), Cydia pomonella (L.), which developed resistance to the biopesticide Cydia pomonella granulovirus (CpGV-M) in Europe. Other studies have shown that, under laboratory conditions, FCM populations differ in their susceptibility to Cryptogran and Cryptex. In order to investigate difference in susceptibility as well as protect against any future resistance by FCM to Cryptogran and Cryptex, a search for novel CrleGV-SA isolates from diseased insects from different geographic regions in South Africa was performed. Six geographic populations (Addo, Citrusdal, Marble Hall, Nelspruit, Baths and Mixed colonies) of FCM were established and maintained in the laboratory. Studies on the comparative biological performance based on pupal mass, female fecundity, egg hatch, pupal survival, adult eclosion and duration of life cycle of the Addo, Citrusdal, Marble Hall, Nelspruit and Mixed colonies revealed a low biological performance for the Citrusdal colony. This was attributed to the fact that FCM populations found in the Citrusdal area are not indigenous and may have been introduced from a very limited gene pool from another region. When insects from five colonies, excluding the Baths colony, were subjected to stress by overcrowding , a latent baculovirus resident in the Addo, Nelspruit, Citrusdal, Marble Hall and Mixed colonies was brought into an overt lethal state. Transmission electron micrographs revealed the presence of GV occlusion bodies (OBs) in diseased insects. DNA profiles obtained by single restriction endonuclease analysis of viral genomic DNA using BamH 1, Sa/1, Xba1 , Pst1, Xh01 , Kpn1, Hindlll and EcoR1 revealed five CrleGV-SA isolates latent within the insect populations. The new isolates were named CrleGV-SA Ado, CrleGV-SA Cit, CrleGV-SA Mbl, CrleGVSA Nels and CrleGV-SA Mix isolates. The novelty of the five CrleGV-SA isolates was confirmed by the presence of unique submolar bands, indicating that each isolate was genetically different. PCR amplification and sequencing of the granulin and egt genes from the five isolates revealed several single nucleotide polymorph isms (SNPs) which, in some cases, resulted in amino acid substitutions. DNA profiles from RFLPs, as well as phylogenetic analysis based on granulin and egt sequencing showed the presence of two CrleGV-SA genome types for the CrleGV-SA isolate. Cryptex and CrleGV-SA Ado, CrleGV-SA Cit, CrleGV-SA Mbl and CrleGV-SA Mix were placed as members of Group one CrleGV-SA, and Cryptogran and CrleGV-SA Nels isolate were placed into Group two CrleGV-SA. In droplet feeding bioassays, the median survival time (STso) for neonate larvae inoculated with Group one and two CrleGV-SA were determined to range from 80 - 88 hours (3.33 - 3.67 days), for all five colonies. LDso values for Group one and two CrleGV-SA against neonates from the Addo, Citrusdal, Marble Hall, Nelspruit and Mixed colonies varied between some populations and ranged from 0.80 - 3.12 OBs per larva, indicating some level of variation in host susceptibility. This is the first study reporting the existence of genetically distinct CrleGV baculovirus isolates infecting FCM in different geographical areas of South Africa. The results of this study have broad-ranging implications for our understanding of baculovirus-host interactions and for the application of baculovirus basedbiopesticides.

This study investigated understorey management in Canterbury, New Zealand, apple orchards for the enhancement of populations of Dolichogenidea tasmanica (Cameron) (Braconidae) for leafroller (Lepidoptera: Tortricidae) biological control. The first objective was to determine the influence of understorey plants on the abundance of D. tasmanica and leafroller parasitism, and to investigate the mechanisms behind this influence. The second was to determine the most suitable understorey plants in terms of their ability to enhance parasitoid abundance, leafroller parasitism, parasitoid longevity, parasitoid fecundity and its ability to not benefit leafroller. Results from three consecutive field trials showed that buckwheat (Fagopyrum esculentum Moench), coriander (Coriandrum sativum L.), alyssum (Lobularia maritima (L.) Desv), and, to a lesser extent, broad bean (Vicia faba L.), enhanced parasitoid abundance and leafroller parasitism. The mechanisms behind the effects of understorey plants had previously been unexplored. However, results here showed that it was the flowers or the buckwheat that 'attracted' the parasitoid to the plant and not the shelter, aphids or microclimate that the plant may also provide. Providing flowering plants in the orchard understorey also increased immigration of parasitoids and enhanced parasitoids and enhanced parasitoid longevity and fecundity in the laboratory. In contrast, the understorey plants had no influence on the female:male ratio of D. tasmanica. Although coriander enhanced leafroller parasitism three-fold in field experiments compared with controls, it failed to enhance the longevity of both sexes of D. tasmanica in the laboratory compared with water-only. Broad bean significantly enhanced parasitoid abundance three-fold and significantly increased parasitism from 0% to 75% compared with the controls on one leafroller release date. However, laboratory trials showed that of male D. tasmancia but it did not enhance female longevity. Also, female D. tasmanica foraging on broad bean produced a total of only three parasitoid cocoons, but this result was based on an overall 6.5% survival of larvae to pupae or to parasitoid cocoon. Furthermore, results suggested that extrafloral nectar secretion decreased as the plants matured. Phacelia (Phacelia tanacetifolia Benth.) did not significantly enhance parasitism rate in the field compared with controls, and numbers of D. tasmanica captured by suction sampling were significantly lower in phacelia treatments compared with alyssum, buckwheat and control plots. Also, laboratory experiments showed that survival of D. tasmanica on phacelia flowers was equivalent to that on water-only and significantly lower than on buckwheat. These results suggest that phacelia does not provide nectar to D. tasmanica, only pollen, and therefore is not a suitable understorey plant for D. tasmanica enhancement in orchards. Buckwheat and alyssum showed the most potential as understorey plants for the enhancement of natural enemies. Buckwheat not only increased numbers of D. tasmanica seven-fold, but also increased numbers of beneficial lacewings (Micromus tasmaniae (Walker)) and hover flies (Syrphidae) captured on yellow sticky traps compared with the controls. It significantly increased leafroller parasitism by D. tasmanica from 0% to 86% compared with the controls (on one date only), and in the laboratory enhanced D. tasmanica longevity and increased fecundity compared with water-only. Similarly, alyssum significantly increased parasitism rate compared with controls, and two-fold more D. tasmanica were suction sampled in these plots compared with controls. It also enhanced longevity of both sexes of D. tasmanica compared with water, and showed the most favourable characteristics in terms of being of no benefit to leafrollers. This is because it was not preferred over apple by leafroller larvae and when they were forced to feed on it, it caused high mortality (94.3%) and low pupal weight (15 mg). Furthermore, alyssum did not enhance the number of fertile eggs produced by adult leafrollers compared with water only. However, further research is required to address the overall effect of buckwheat and alyssum on crop production and orchard management, including effects on fruit yield and quality, frost risk, disease incidence, soil quality, weeds and other pests. Also, research into the ability of these plants to survive in the orchard with little maintenance, and into the optimal sowing rates, would be useful. Sampling natural populations of leafroller within each treatment showed that damage from leafrollers and the number of leafroller larvae were respectively 20.3% and 29.3% lower in the flowering treatments compared with the controls. Furthermore, field trials showed up to a six-fold increase in leafroller pupae in controls compared with buckwheat and alyssum. This suggests that increasing leafroller parasitism rate from understorey management in orchards will translate into lower pest populations, although neither larval numbers/damage nor pupal numbers differed significantly between treatments. Trapping D. tasmanica at a gradient of distances showed that this parasitoid travels into rows adjacent to buckwheat plots, indicating that growers may be able to sow flowering plants in every second or third row of the orchard, and still enhance leafroller biocontrol while minimising the adverse effects of a cover crop. Sowing buckwheat and alyssum in orchard understoreys may enhance biological control of apple pests in organic apple production and reduce the number of insect growth regulators applied in IFP programmes. However, the challenge still remains to investigate whether conservation biological control can reduce leafroller populations below economic thresholds.

In this study, buckwheat (Fagopyrum esculentum Moench) and alyssum (Lobularia maritima (L.)) flowers were used to examine the effect of floral resources on the efficacy of the leafroller parasitoid Dolichogenidea tasmanica (Cameron) in vineyards. This was done by assessing the influence of these flowers on parasitoid abundance and parasitism rate, and by investigating the consequences of this for leafroller abundance. In laboratory experiments, alyssum flowers were used to investigate the effect of floral food on the longevity, fecundity and sex ratio of D. tasmanica. Dolichogenidea tasmanica comprised more than 95 % of parasitoids reared from field collected leafrollers in this study. The abundance of D. tasmanica during the 1999-2000 growing season was very low compared with previous studies, possibly due to the very low abundance of its leafroller hosts during the experiment. The number of males of this species on yellow sticky traps was increased (although not significantly) when buckwheat flowers were planted in a Marlborough vineyard; however, the number of female D. tasmanica on traps was no greater with flowers than without. The abundance of another leafroller parasitoid, Glyptapanteles demeter (Wilkinson)(Hymenoptera: Braconidae), on traps was also not significantly affected by the presence of buckwheat flowers, although females of this species were caught in greater numbers in the control than in buckwheat plots. Naturally-occurring leafrollers were collected from three vineyard sites in Marlborough, and one in Canterbury during the 2000-2001 season to assess the effect of buckwheat and alyssum flowers on parasitism rate. Parasitism rate more than doubled in the presence of buckwheat at one of the Marlborough vineyards, but alyssum had no effect on parasitism rate in Canterbury. A leafroller release/recover method, used when naturally-occurring leafrollers were too scarce to collect, was unable to detect any effect of buckwheat or alyssum on parasitism rate. Mean parasitism rates of approximately 20 % were common in Marlborough, although rates ranged from 0 % to 45 % across the three vineyard sites in that region. In Canterbury in April, mean parasitism rates were approximately 40 % (Chapter 4). Rates were higher on upper canopy leaves (40-60 %) compared with lower canopy leaves and bunches (0-25 %). Leafroller abundance was apparently not affected by the presence of buckwheat in Marlborough, or alyssum in Canterbury. Buckwheat did, however, significantly reduce the amount of leafroller evidence (webbed leafroller feeding sites on leaves or in bunches) in Marlborough, suggesting that the presence of these flowers may reduce leafroller populations. Leafrollers infested less than 0.1 % of Cabernet Sauvignon leaves throughout the 1999-2000 growing season, but increased in abundance in bunches to infest a maximum of 0.5 % of bunches in late March in Marlborough. In Pinot Noir vines in the 2000-2001 season, leafroller abundance was also low, although sampling was not conducted late in the season when abundance reaches a peak. In Riesling vines in Canterbury, between 1.5 % and 2.5 % of bunches were infested with leafrollers in April. In the laboratory, alyssum flowers significantly increased the longevity and lifetime fecundity of D. tasmanica compared with a no-flower treatment. However, daily fecundity was not increased by the availability of food, suggesting that the greater lifetime fecundity was related to increases in longevity. Parasitoids were also able to obtain nutrients from whitefly honeydew, which resulted in similar longevity and daily fecundity to those when alyssum flowers were present. The availability of food had a significant effect on the offspring sex ratio of D. tasmanica. Parasitoids reared from naturally-occurring leafrollers produced an equal sex ratio, assumed to be the evolutionarily stable strategy (ESS) for this species. In the laboratory, this ESS was observed only when parasitoids had access to alyssum flowers. Without food, or with honeydew only, sex ratios were strongly male-biased. In the field, floral resources affected the sex ratio of D. tasmanica only when this species was reared from leafrollers released and recovered in Marlborough. In that experiment, buckwheat shifted the sex ratio in favour of female production from the equal sex ratio found in control plots. No firm explanations can be given to account for these results, due to a lack of research in this area. Possible mechanisms for the changes in sex ratio with flowers are discussed. This study demonstrated that flowers are an important source of nutrients for D. tasmanica, influencing the longevity, fecundity and offspring sex ratio of this species. However, only some of the field experiments were able to show any positive effect of the provision of floral resources on parasitoid abundance or parasitism rate. More information is needed on the role these parasitoids, and other natural enemies, play in regulating leafroller populations in New Zealand vineyards, and on how they use floral resources in the field, before recommendations can be made regarding the adoption of this technology by growers.

Le maïs cultivé en zones de savanes de Côte d'Ivoire est attaqué par 5 espèces de Lépidoptères foreurs qui s'attaquent soit a la tige, soit a l'épi, soit aux deux. Étude de la chétotaxie larvaire des différents foreurs, de l'habitus des stades préimaginaux, de la morphologie et de la biologie de Mussidia nigrivenella. Résultats de l'étude des fluctuations des populations de ces foreurs. Mise en évidence de l'efficacité de la deltaméthrine en concentré émulsifiable a 15 grammes de matière active à l'hectare. Iimpact des divers insectes sur le rendement et calcul d'équations de régression reliant la perte de rendement a l'attaque

There is currently little knowledge about the dynamics of invertebrates in Australian viticultural ecosystems. This study was conducted in Coonawarra vineyards over three seasons (years) and has focused on identifying natural enemies, their seasonal phenology, multiple species interactions, and potential for the suppression of the pest lepidopteran Epiphyas postvittana (Tortricidae). The work presented in this thesis shows that endemic natural enemies have far greater potential to control E. postvittana than has been realised. An initial survey identified a diverse and abundant range of potential natural enemies. Of these, the species most likely to attack E. postvittana include a predatory mite Anystis baccarum and a number of hymenopteran parasitoids. The most abundant parasitoid in the vineyards was a braconid, Dolichogenidea tasmanica. Understanding the characteristic behaviour of parasitoids in response to host density can help to gauge their potential for pest suppression. The results of large-scale field experiments showed that the response of D. tasmanica to the density of E. postvittana was inversely density-dependent, and that parasitism was consistently higher in Cabernet Sauvignon compared with Chardonnay varieties. Despite the fact that interactions among multiple species of natural enemies can increase or decrease pest suppression, particularly when they share a common prey/host, few multispecies interactions have been investigated. Laboratory studies identified a novel interaction between the predatory mite A. baccarum an abundant predator in the vine canopy, the parasitoid D. tasmanica and host E. postvittana larvae. Although A. baccarum readily ate E. postvittana eggs and free roaming larvae, they could not access larva in their silk leaf rolls. However, the addition of D. tasmanica significantly increased predation of E. postvittana larvae, by altering the behaviour of host larvae and increasing their vulnerability to the mite. Experiments conducted at a landscape level in the Coonawarra showed that D. tasmanica was also present in habitat other than vineyards including native vegetation. However, it was not present in highly disturbed habitats. Although the exact mechanism for this remains unknown, results indicate that viticultural practices and resources in the surrounding landscape can influence the presence of parasitoids. Together, the findings presented in this thesis make a significant contribution towards developing sustainable pest management in Australian viticulture.
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Thesis (Ph.D.) -- University of Adelaide, School of Earth and Environmental Sciences, 2008

There is currently little knowledge about the dynamics of invertebrates in Australian viticultural ecosystems. This study was conducted in Coonawarra vineyards over three seasons (years) and has focused on identifying natural enemies, their seasonal phenology, multiple species interactions, and potential for the suppression of the pest lepidopteran Epiphyas postvittana (Tortricidae). The work presented in this thesis shows that endemic natural enemies have far greater potential to control E. postvittana than has been realised. An initial survey identified a diverse and abundant range of potential natural enemies. Of these, the species most likely to attack E. postvittana include a predatory mite Anystis baccarum and a number of hymenopteran parasitoids. The most abundant parasitoid in the vineyards was a braconid, Dolichogenidea tasmanica. Understanding the characteristic behaviour of parasitoids in response to host density can help to gauge their potential for pest suppression. The results of large-scale field experiments showed that the response of D. tasmanica to the density of E. postvittana was inversely density-dependent, and that parasitism was consistently higher in Cabernet Sauvignon compared with Chardonnay varieties. Despite the fact that interactions among multiple species of natural enemies can increase or decrease pest suppression, particularly when they share a common prey/host, few multispecies interactions have been investigated. Laboratory studies identified a novel interaction between the predatory mite A. baccarum an abundant predator in the vine canopy, the parasitoid D. tasmanica and host E. postvittana larvae. Although A. baccarum readily ate E. postvittana eggs and free roaming larvae, they could not access larva in their silk leaf rolls. However, the addition of D. tasmanica significantly increased predation of E. postvittana larvae, by altering the behaviour of host larvae and increasing their vulnerability to the mite. Experiments conducted at a landscape level in the Coonawarra showed that D. tasmanica was also present in habitat other than vineyards including native vegetation. However, it was not present in highly disturbed habitats. Although the exact mechanism for this remains unknown, results indicate that viticultural practices and resources in the surrounding landscape can influence the presence of parasitoids. Together, the findings presented in this thesis make a significant contribution towards developing sustainable pest management in Australian viticulture.
Thesis (Ph.D.) -- University of Adelaide, School of Earth and Environmental Sciences, 2008