Efeito de diferentes pesticidas sobre os ácaros predadores Phytoseiulus macropilis e Amblyseius idaeus (Acari: Phytoseiidae).

Entre julho de 1991 e junho de 1992 foram realizados no laboratorio de Entomologia do CNPDA, testes de seletividade dos produtos Malation, Triclorfon, Mancozeb, Cardaril, Deltametrina, Enxofre, Permetrina, Oxicloreto de cobre, Bacillus thuringiensis, Pirimicarbe, Carbamoil e Benomil aos ácaros predadores fitoseídeos Phytoseiulus macropilis (Banks) e Amblyseius idaeus (Demark & Muma). Estes testes foram realizados utilizando-se o método de imersão de laminas. Em cada lamina foram fixados 20 ácaros predadores sobre fita adesiva com dupla face. Com cada produto foram preparadas soluções com diferentes concentrações: x (dose recomendada); 0,0 x (testemunha, H2O destilada); 0,01x; 0,1x; 10x e 100x. As lâminas foram mergulhadas durante 5 segundos nas soluções e as leituras realizadas 5 e 24 horas após. O produto que apresentou maior toxicidade a P. macropilis foi o piretroide Permetrina, que causou mortalidade de 100% dos predadores, em todas as doses, 24 horas após o tratamento. O produto que apresentou menor toxicidade foi o enxofre. O produto de maior toxicidade a A. idaeus foi o piretróide Deltametrina que provocou mortalidade de 100% em todas as doses, 5 horas após o tratamento. O produto que causou menor toxicidade a esse predador foi também o enxofre. Os resultados obtidos neste ensaio indicam que aplicações de piretróides não são recomendáveis no manejo integrado do ácaro rajado com uso daqueles fitoseídeos.

Controle biológico do ácaro rajado por ácaros predadores fitoseídeos (Acari: Tetranychidae, Phytoseiidae) em pepino.

Em janeiro de 1992 foi instalado no campo experimental do CNPDA, em Jaguariúna/SP, um experimento para testar a viabilidade técnica do controle biológico do ácaro rajado (Tetranychus urticae Koch) por ácaros predadores fitoseídeos. O delineamento experimental utilizado foi de blocos casualizados com 4 repetições e 5 tratamentos; T1= liberação de 10 ácaros rajados/planta + Malation (para eliminacao dos predadores nativos); T2= 10 ácaros rajados /planta; T3= 10 acaros rajados + 8 ácaros predadores Phytoseiulus macropilis (Banks)/planta; T4= 10 ácaros rajados + 8 ácaros predadores Amblyseius idaeus (Denmark & Muma)/planta; T5= Avermectina-testemunha sem liberação de ácaros. Esses tratamentos foram aplicados em 20 plantas tomadas ao acaso na área útil de cada parcela. Antes da liberação dos predadores não havia diferença significativa entre os tratamentos, exceto em relação a testemunha T5. Após a liberação dos predadores, destacou-se o tratamento T4 que apresentou o número de ácaros rajados significativamente menor em relação aos tratamentos T1, T2, T3. Assim, o predador A. idaeus foi capaz de controlar a população do ácaro rajado, mantendo-a em niveis inferiores em relação aos demais tratamentos (exceto T5). Esse predador esteve presente nas plantas de pepino desde a sua liberação ate o término do experimento. Já a população de P. macropilis extingui-se 10 dias após a liberação.

Controle biológico do ácaro rajado com ácaros predadores fitoseideos (Acari: Tetranychidae, Phytoseiidae) em culturas de pepino e morango.

O ácaro rajado, Tetranychus urticae Koch, é considerado uma das principais pragas de hortaliças e varias outras culturas no Brasil, em áreas onde um considerável volume de acaricidas e anualmente empregado no seu controle. O objetivo deste trabalho foi testar a viabilidade técnica do empregado dos fitoseideos Amblyseius idaeus (Denmark & Muma) e Phytoseiulus macropilis (Banks), comumente encontrados no Brasil, no controle de T. urticae em pepino e morangueiro em Jaguariúna, SP. Utilizou-se o delineamento experimental em blocos casualizados, com 4 repetições e 4 tratamentos (T1-T4) para o pepino e 5 tratamentos para o morangueiro (T1-T5): T1, testemunha; T2 pulverização semanais de Malation para a exclusão de predadores nativos; T3, liberação periodicos de A. idaeus; T4, liberações periódicas de P.macropilis; T5, pulverizações semanais de avermectina para a exclusão de ácaros fitófagos e predadores nativos. Apenas A. idaeus se estabeleceu na cultura do pepino, reduzindo significativamente a população de T. urticae. Ambas espécies de predadores se estabeleceram na cultura do morango e reduziram significativamente a população de T. urticae.

Vadelman (Rubus idaeus) kasvunsäätö proheksadioni-kalsiumin avulla

Vadelman (Rubus idaeus L.) viljelyala on kasvanut Suomessa voimakkaasti viimeisten viiden vuoden aikana. Vuonna 2005 vadelmaa viljeltiin 473 hehtaarilla. Vadelman satoindeksi on alhainen. Sen pääverso on pitkä, mikä hankaloittaa hoitotoimenpiteitä. Kasvin satoisimmat silmut ovat pääverson kärjessä, joten latvontaa ei kannata tehdä. Suomessakin haluttaisiin kehittää vadelman kasvihuonetuotantoa satokauden pidentämiseksi. Kasvihuonetuotannossa vegetatiivisen kasvun säätäminen on vielä tärkeämpää kuin avomaan viljelyssä. Proheksadioni-kalsium (ProCa) on gibberelliinisynteesi-inhibiittori, jonka on havaittu lyhentävän nivelvälejä sekä lisäävän versojen ja lehtien hiilihydraattipitoisuutta mm. omenalla (Malus domestica Borkh.). Tämän Helsingin yliopiston soveltavan biologian laitoksella toteutetun tutkimuksen tarkoituksena oli selvittää, miten ProCa vaikuttaa kahden kesävadelmalajikkeen ensimmäisen vuoden versojen vegetatiiviseen kasvuun ja edelleen sadontuottokykyyn. Tässä pro gradu -työssä tutkittiin ProCa:n vaikutusta kahden kesävadelmalajikkeen vegetatiiviseen kasvuun, satopotentiaaliin sekä versojen ja silmujen hiilihydraattipitoisuuksiin kesällä 2005 ja talvella 2006. Koetaimina oli 120 Tulameen-lajikkeen ja 150 Glen Ample -lajikkeen tainta. Taimet käsiteltiin kesäkuussa ja käsittely uusittiin osalle taimista neljän viikon kuluttua ensimmäisestä käsittelystä. Koekäsittelyitä oli neljä: ProCa 100 ppm, ProCa 200 ppm, ProCa 100 ppm + ProCa 100 ppm ja ProCa 200 ppm + ProCa 200 ppm. Kontrollikäsittelyitä oli kaksi: vesi sekä vesi + vesi. Kesän aikana havainnoitiin pääverson pituuskasvua ja nivelien määrää. Kesän kasvatuskauden päätyttyä Glen Ample -lajikkeen silmujen ja versojen liukoiset hiilihydraatit ja tärkkelys määritettiin entsymaattisesti. Kylmävarastoinnin jälkeen (71 vrk) tammikuussa 2006 taimet siirrettiin hyötöön kasvihuoneeseen. Hyödön aikana havainnoitiin silmujen puhkeamista ja kukintaa. Hyödön päättyessä hanka- ja pääversojen kuivapaino sekä hankaversojen pituus mitattiin. Kesän 2005 aikana ProCa lyhensi uusien nivelvälien pituuksia väliaikaisesti 1 4 viikon kuluttua käsittelystä. Nivelien lukumäärässä ei tapahtunut suuria muutoksia. Tulameen-lajikkeella kasvu kiihtyi käsittelyn vaikutuksen loputtua. Nivelvälien lyhentymisen takia pääverson kokonaispituus jäi kontrolliin verrattuna 35 % lyhyemmäksi Glen Ample -lajikkeella (ProCa 200 + ProCa 200) ja 14 % lyhyemmäksi Tulameen-lajikkeella samalla käsittelyllä. Myös käsittely ProCa 200 vähensi pääverson pituutta 6 % kontrolliin verrattuna Tulameen-lajikkeella. ProCa-käsittely lisäsi versojen ja silmujen hiilihydraattipitoisuuksia, mutta tuloksissa oli paljon hajontaa. Glen Ample -lajikkeella kaksinkertaiset käsittelyt vähensivät puhjenneiden silmujen osuutta 8 15 päivän kuluttua hyödön aloittamisesta 24 29 %, mutta käsittelyiden väliset erot tasoittuivat hyödön loppuun mennessä. Lisäksi kaksinkertaiset ProCa-käsittelyt vähensivät kukkien määrää 29 44 % 33 65 päivää hyödön alusta, mutta erot tasoittuivat hyödön loppuun mennessä. Käsittelyt eivät vaikuttaneet Tulameen-lajikkeen silmujen puhkeamiseen ja kukintaan. Satopotentiaalissa ei ollut eroa ProCa-käsittelyiden välillä kummallakaan lajikkeella. Glen Ample -lajikkeella hankaversojen kokonaismäärä väheni 27 32 % kasveilla, joita oli käsitelty kaksi kertaa ProCa:lla. Hankaversot olivat kuitenkin vastaavasti 29 43 % pidempiä. Tulameen-lajikkeella ProCa 100 -käsittely vähensi hankaversojen määrää 15 %, mutta hankaversojen pituuteen käsittelyillä ei ollut vaikutusta. Käsittelyillä ei ollut vaikutusta hankaversojen nivelvälien pituuteen kummallakaan lajikkeella. Tutkimuksen perusteella kesällä annetulla ProCa-käsittelyllä voidaan lyhentää pääverson pituutta, mutta satopotentiaaliin käsittelyllä ei ole vaikutusta. Käsittelyajankohta on kriittinen, koska kukkasilmujen kehitys näyttäisi häiriintyvän, jos ruiskutus tehdään loppukesällä.

Mehiläisten (Apis mellifera) kukkavierailut mansikalla (Fragaria x ananassa) ja vadelmalla (Rubus idaeus) sekä kukkavierailuihin vaikuttavat tekijät

Hyönteispölytys lisää monien ristipölytteisten viljelykasvien siemensatoa sekä parantaa sadon laatua. Marjakasveilla, kuten mansikalla ja vadelmalla marjojen koko suurenee sekä niiden laatu paranee onnistuneen pölytyksen seurauksena. Aiempien havaintojen mukaan mansikan kukat eivät pääsääntöisesti houkuttele mehiläisiä, kun taas vadelma on yksi mehiläisten pääsatokasveista. Tutkimuksen tarkoituksena oli selvittää, miten tehokkaasti mehiläiset vierailevat mansikalla sekä vadelmalla, keskittyen kukkakohtaisiin käynteihin tuntia kohti. Mehiläisiä voidaan käyttää Gliocladium catenulatum-vektoreina torjuttaessa mansikan ja vadelman harmaahometta (Botrytis cinerea). Kukkavierailujen perusteella arvioidaan, onko vektorilevitys riittävän tehokas torjumaan harmaahometta ja miten hyvin mehiläisiä voidaan käyttää pölytyspalveluihin, etenkin mansikalla. Havainnot kerättiin kuudelta eri tilalta Sisä-Savosta kesällä 2007. Kukkavierailuja laskettiin mansikan ja vadelman kukinnan aikana erilaisissa sääolosuhteissa, eri kellonaikoina ja eri etäisyyksillä mehiläispesistä. Kukat valittiin satunnaisesti, ja valintaperusteena oli kukan avonaisuus. Tarkkailuaika riippui mehiläisten lentoaktiivisuudesta. Mansikan koko havaintojakson keskiarvoksi tuli 1,75 käyntiä kukkaa kohti tunnissa. Vadelmalla vastaava luku oli 4,27, joten keskiarvojen perusteella vadelma oli houkuttelevampi kuin mansikka. Kasvukauden vaiheella ei ollut eroja vierailuihin kummallakaan kasvilla, mutta vuorokaudenajan suhteen vierailuja oli enemmän aamupäivällä kuin iltapäivällä. Lämpötila korreloi positiivisesti vierailutiheyden kanssa kummallakin kasvilla. Sääolosuhteet rajoittivat havaintojen keräämistä ja kesä oli erittäin sateinen. Mehiläiset vierailivat kukissa riittävästi haastavissakin sääolosuhteissa niin, että harmaahometorjunta onnistui. Vektorilevitystä suunnitellessa, etenkin mansikalla, tulee ottaa huomioon pesien sijoittelu sekä riittävä lukumäärä. Pesien ravinnontarpeen tulee olla suuri, jotta mehiläiset keräisivät ravintoa kukista mahdollisimman tehokkaasti. Pesiin voidaan lisätä tarvittaessa avosikiöitä tai poistaa siitepölyvarastoja ravinnonkeruuaktiivisuuden lisäämiseksi. Lisätutkimusta tarvitaan pesien sijoittelun, kilpailevien kasvien sekä mansikkalajikkeiden houkuttelevuuden vaikutuksesta vierailutiheyteen. Suomalaisten mansikkalajikkeiden meden sekä siitepölyneritystä olisi myös hyvä selvittää.

Évaluation de suppléments alimentaires pour deux espèces d’acariens prédateurs, Amblyseius swirskii et Neoseiulus cucumeris (Acari: Phytoseiidae) pour l’optimisation du contrôle biologique du thrips des petits fruits (Frankliniella occidentalis) en serriculture

Projet réalisé en cotutelle avec Jacques Brodeur et Les Shipp

Side effects of modern pesticides on adults of the predatory mite Amblyseius swirskii (Athias-Henriot) (Acari: Phytoseiidae) under laboratory conditions

Amblyseius swirskii (Athias-Henriot) is a polyphagous predatory mite which feeds on pollen and small arthropod preys like whiteflies, thrips and mites. This species is widely used in IPM programs in greenhouses, being essential for its success, to obtain information about the non target effects of the pesticides currently used in those crops where the mite is artificially released. This work describes a laboratory contact residual test for evaluating lethal (mortality after 72 hour exposure to fresh residues) and sublethal effects (fecundity and fertility of the surviving mites) of eleven modern pesticides to adults of A. swirskii. Spiromesifen is lipogenesis inhibitor; flonicamid a selective feeding inhibitor with a mode of action not totally known; flubendiamide a modulator of the rhyanodin receptor, sulfoxaflor has a complex mode of action not totally ascertained; metaflumizone is a voltage dependent sodium channel blocker; methoxyfenozide is an IGR, spirotetramat inhibits lipids; abamectin and emamectin activate the Cl- channel; spinosad is a neurotix naturalyte and deltamethrin a pyrethroid used as positive standard. Selected pesticides are effective against different key pests present in horticultural crop areas and were always applied at the maximum field recommended concentration in Spain if registered, or at the concentration recommended by the supplier. Out of the tested pesticides, spiromesifen, flonicamid, flubendiamide, sulfoxaflor, metaflumizone, methoxyfenozide and spirotetramat were harmless to adults of the predatory mite (IOBC toxicity class 1). The rest of pesticides exhibited some negative effects: emamectin was slightly harmful (IOBC 2), deltamethrin moderately harmful (IOBC 3) and spinosad and abamectin harmful (IOBC 4). Further testing under more realistic conditions is needed for those pesticides having some harmful effect on the mite prior deciding their joint use or not. Key words: Amblyseius swirskii, adults, laboratory, residual test, spiromesifen, flonicamid, flubendiamide, sulfoxaflor, metaflumizone, methoxyfenozide, spirotetramat, emamectin, deltamethrin, abamectin, spinosad.

Compatibilidad de Eretmocerus mundus Mercet (Hymenoptera: Aphelinidae) y Amblyseius swirskii Athias-Henriot (Acari: Phytoseiidae), importantes enemigos naturales de la mosca blanca Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) en cultivos hortícolas, con nuevas barreras físicas selectivas y modernos plaguicidas

Los programas de Gestión Integrada de Plagas (GIP) promueven el uso de estrategias de control que sean respetuosas con el medio ambiente, sin embargo el uso de insecticidas en los cultivos hortícolas sigue siendo necesario para el control de determinadas plagas, como es el caso de la mosca blanca Bemisia tabaci (Gennadius). Por ello, el objetivo de esta tesis es el estudio de la integración de las tres estrategias de control más empleadas hoy en día para el control de plagas: el control biológico, el físico y el químico. Una primera parte de este trabajo ha consistido en el estudio de los efectos letales y subletales de once insecticidas, aplicados a la dosis máxima de campo, sobre los enemigos naturales Eretmocerus mundus Mercet y Amblyseius swirskii Athias-Henriot, mediante ensayos de laboratorio y persistencia (laboratorio extendido). Para la evaluación de la toxicidad de los insecticidas sobre los estados de vida más protegidos de estos enemigos naturales, se trataron bajo la Torre de Potter las pupas de E. mundus y los huevos de A. swirskii. Además, se llevaron a cabo ensayos de contacto residual para determinar los efectos letales y subletales de estos insecticidas sobre el estado adulto de ambas especies de enemigos naturales. Para ello, los pesticidas se aplicaron sobre placas de cristal (laboratorio) o sobre plantas (laboratorio extendido: persistencia). Los resultados mostraron que los insecticidas flonicamida, flubendiamida, metaflumizona, metoxifenocida, spiromesifen y spirotetramat eran compatibles con el estado de pupa de E. mundus (OILB 1: Inocuos). Sin embargo, abamectina, deltametrina y emamectina fueron categorizadas como ligeramente tóxicas (OILB 2) al causar efectos deletéreos. Los dos pesticidas más tóxicos fueron spinosad y sulfoxaflor, los cuales redujeron significativamente la emergencia de las pupas tratadas (OILB 4: Tóxicos). Flonicamida, flubendiamida, metoxifenocida y spiromesifen fueron compatibles con el estado adulto de E. mundus (OILB 1: Inocuos). Abamectina, deltametrina, emamectina, metaflumizona y spiromesifen pueden ser recomendados para su uso en programas de GIP, si se usan los plazos de seguridad apropiados, de acuerdo con la persistencia de cada uno de estos insecticidas, antes de la liberación del enemigo natural. Al contrario, spinosad y sulfoxaflor no resultaron ser compatibles (OILB D: Persistentes), aunque la realización de ensayos adicionales es necesaria para ver los efectos de los mismos en campo. Todos los insecticidas estudiados, excepto el spirotetramat (OILB 2: Ligeramente tóxico), fueron selectivos para el estado de huevo de A. swirskii (OILB 1: Inocuos). Flonicamida, flubendiamida, metaflumizona, metoxifenocida, spiromesifen, spirotetramat y sulfoxaflor, fueron compatibles con el estado adulto de A. swirskii (OILB 1: Inocuos). Abamectina, deltametrina, emamectina y spinosad pueden ser recomendados para su uso en programas de GIP, si se usan los plazos de seguridad apropiados, de acuerdo con la persistencia de cada uno de estos insecticidas, antes de la liberación del enemigo natural. Entre las nuevas estrategias de la GIP, los plásticos y mallas fotoselectivas han demostrado ser una herramienta importante para el control de plagas y enfermedades en cultivos hortícolas protegidos. Por ello, en una segunda parte de este trabajo, se estudiaron tanto los efectos directos, como la combinación de efectos directos y mediados por planta y plaga de ambientes pobres en luz UV, en presencia o ausencia del Virus del rizado amarillo del tomate (TYLCV), sobre E. mundus. En primer lugar, se realizó un ensayo al aire libre para la evaluación de la capacidad de vuelo de E. mundus en cajas tipo túnel (1 x 0,6 x 0,6 m) cubiertas con distintas barreras absorbentes de luz UV. Se detectó un efecto directo en la capacidad de orientación de E. mundus, debido a que este parasitoide utiliza estímulos visuales para localizar a sus huéspedes, únicamente en las barreras que bloqueaban más del 65% de la luz UV (malla G). En segundo lugar, bajo condiciones de invernadero, se evaluó la combinación de efectos directos y mediados por planta y plaga sobre E. mundus, usando plantas de tomate sanas o infectadas con el TYLCV y cajas (30 x 30 x 60 cm) cubiertas con los distintos plásticos fotoselectivos. En este caso, no se observó ningún efecto en la capacidad benéfica del parasitoide cuando este estaba en contacto con plantas de tomate infestadas con ninfas de B. tabaci, lo que demuestra que este insecto usa estímulos táctiles para encontrar a sus huéspedes a cortas distancias. Además, las diferentes condiciones de radiación UV estudiadas tuvieron cierto impacto en la morfología, fisiología y bioquímica de las plantas de tomate, infestadas o no con el virus de la cuchara, detectándose pequeñas alteraciones en alguno de los parámetros estudiados, como el peso fresco y seco, el contenido en H y el espesor de las cutículas y de las paredes celulares de la epidermis foliar. Por último, no se observaron efectos de la radiación UV mediados por planta, ni en B. tabaci ni en su parasitoide, E. mundus. En una tercera parte, se evaluaron los efectos de una malla tratada con bifentrin sobre ambos enemigos naturales, en ensayos de laboratorio, semicampo y campo. Las mallas tratadas fueron diseñadas originariamente para el control de mosquitos vectores de la malaria, y actualmente se está trabajando para su uso en agricultura, como una nueva estrategia de control de plagas. En ensayos de laboratorio, cuando adultos de E. mundus y A. swirskii se expusieron por contacto durante 72 horas con la malla tratada (cajas de 6 cm diámetro), se registró una alta mortalidad. Sin embargo, en el ensayo de preferencia, estos enemigos naturales no fueron capaces de detectar la presencia de bifentrin y, en aquellos individuos forzados a atravesar la malla tratada, no se observó mortalidad a corto plazo (72 horas). En estudios de semicampo, llevados a cabo bajo condiciones de invernadero en cajas de 25 x 25 x 60 cm de altura, la capacidad benéfica de E. mundus no se vio afectada. Finalmente, en ensayos de campo llevados a cabo en invernaderos comerciales (4000m2) en Almería, A. swirskii no se vio afectado por la presencia en el cultivo de la malla tratada con bifentrin y los niveles de infestación de B. tabaci y F. occidentalis detectados bajo dicha malla, fueron inferiores a los del control. Por último, se ha evaluado la composición de la microflora bacteriana de tres especies de parasitoides, E. mundus, Eretmocerus eremicus Rose & Zolnerowich y Encarsia formosa Gahan, y la influencia de la misma en su susceptibilidad a insecticidas. Se llevó a cabo una extracción total de ADN de los insectos y la región variable V4 del ARNr se amplificó usando cebadores universales bacterianos. Para identificar las secuencias de los géneros bacterianos presentes en los parasitoides, se realizó una Next Generation sequencing (Illumina sequencing). Una vez identificados los géneros bacterianos, el gen ADNr 16S de las Actinobacterias se amplificó del ADN extraído de los insectos, usando cebadores universales bacterianos y específicos de Actinobacterias, y los productos de la Nested PCR fueron clonados para identificar todas las especies del género Arthrobacter. Tres bacterias (A. aurescens Phillips, A. nicotinovarans Kodama, Yamamoto, Amano and Amichi y A. uratoxydans Stackebrandt, Fowler, Fiedler and Seiler), próximas a las especies de Arthrobacter presentes en los parasitoides, se obtuvieron de la colección bacteriana del BCCMTM/LMG y se midió su actividad esterasa. Finalmente, se realizaron ensayos con antibióticos (tetraciclina) y de contacto residual con insecticidas (abamectina) para determinar la influencia de las especies de Arthrobacter en la susceptibilidad de E. mundus a insecticidas. Los resultados muestran que este género bacteriano puede afectar a la toxicidad de E. mundus a abamectina, mostrando la importancia de la comunidad microbiana en enemigos naturales, factor que debe ser considerado en los estudios de evaluación de los riesgos de los insecticidas. ABSTRACT Integrated Pest Management (IPM) programs promote the use of control strategies more respectful with the environment; however the use of insecticides in vegetable crops is still needed to control certain pests, such as the whitefly Bemisia tabaci (Gennadius). Therefore, the objective of this work is to study the integration of the three most commonly used pest control strategies nowadays: biological, physical and chemical control. Firstly, the lethal and sublethal effects of eleven insecticides, applied at their maximum field recommended concentration, on the parasitic wasp Eretmocerus mundus Mercet and the predator Amblyseius swirskii Athias-Henriot has been assessed in the laboratory and in persistence tests (extended laboratory). To test the effects of pesticides on the most protected life stage of these natural enemies, E. mundus pupae and A. swirskii eggs were sprayed under a Potter precision spray tower. Laboratory contact tests were therefore conducted to determine the lethal and sublethal effects of these pesticides on the adult stage of these natural enemies. In the residual contact tests the pesticides were applied on glass plates (laboratory) or plants (extended laboratory: persistence). The study showed that the insecticides flonicamid, flubendiamide, metaflumizone, methoxyfenozide, spiromesifen and spirotetramat were selective for E. mundus pupae (IOBC 1: Harmless). Nevertheless, abamectin, deltamethrin and emamectin were categorized as slightly harmful (IOBC 2) due to the deleterious effects caused. The two most harmful pesticides were spinosad and sulfoxaflor, which significantly reduced the adult emergence from treated pupae (IOBC 4: Harmful). Flonicamid, flubendiamide, methoxyfenozide and spiromesifen were compatible with E. mundus adults (IOBC 1: Harmless). Base on the duration of the harmful activity, abamectin, deltamethrin, emamectin, metaflumizone and spirotetramat could be recommended for use in IPM programs if appropriate safety deadlines are used before the natural enemy release. On the contrary, spinosad and sulfoxaflor were not compatible (IOBC D: persistent), although additional studies are required to determine their effects under field conditions. All the pesticides tested, except spirotetramat (IOBC 2: Slightly harmful), were selective for A. swirskii eggs (IOBC 1: Harmless). Flonicamid, flubendiamide, metaflumizone, methoxyfenozide, spiromesifen, spirotetramat and sulfoxaflor were compatible with A. swirskii adults (IOBC 1: Harmless). However, abamectin, deltamethrin, emamectin and spinosad could be recommended for use in IPM programs if appropriate safety deadlines are used before the natural enemy release. Among new IPM strategies, UV-absorbing photoselective plastic films and nets have been shown to be an important tool for the control of pests and diseases in horticultural protected crops. Because of that, we secondly studied the plant and pest insect-mediated and/or the direct effects on E. mundus under different UV radiation conditions, in presence or absence of the Tomato Yellow Leaf Curl Virus (TYLCV). In the first experiment, performed outdoors, the flight activity of E. mundus was studied in one-chamber tunnels (1 x 0.6 x 0.6 m) covered with different photoselective barriers. Because E. mundus uses visual cues for host location at a long distance, a direct effect on its host location ability was detected, but only in the UV-absorbing barriers blocking more than 65% of the UV light (G net). In a second experiment, the direct and plant and pest insect-mediated effects of different UV radiation conditions on E. mundus were studied, inside cages (30 x 30 x 60 cm) covered with the different UVplastic films and under greenhouse conditions, using healthy or TYLCV-virus infected tomato plants. In this case, not any effect on the beneficial capacity of this parasitoid was detected, proving that he uses tactile cues at a short distance of the host. Moreover, the different UV radiation conditions studied had a certain direct impact in the morphology, physiology and biochemistry of tomato plants infested or not with the TYLCV, and small alterations in some parameters such as fresh and dry weight, H percentage and cuticle and cell wall thickness of epidermal cells of the leaves, were detected. Finally, none plant-mediated UV effects neither in the whitefly B. tabaci nor in their parasitic wasp were found. Thirdly, the effects of a bifenthrin treated net were evaluated in different laboratory, semi-field and field experiments on the natural enemies studied. Treated nets were developed long time ago aiming at the control of the mosquitoes vectors of malaria, and nowadays, there is a great interest on assessing the possibility of their use in agriculture. In laboratory assays, a high mortality was recorded when E. mundus and A. swirskii adults were exposed by contact to the bifenthrin treated net for 72 hours in small cages (12 cm diameter). However, these natural enemies were not able to detect the presence of bifenthrin in a dual-choice test and no short-term mortality (72 hours) was recorded in those individuals that went through the treated net. In semi-field assays, performed under greenhouse conditions with cages of 25 x 25 x 60 cm high, the beneficial capacity of E. mundus was not affected. Finally, in field assays carried out in commercial multispan greenhouses (4000 m2) in Almería, A. swirskii was not affected by the presence of the bifenthrin treated net in the crop and the B. tabaci and F. occidentalis infestation levels were significantly lower than in the control. Finally, the composition of the microflora present in three species of parasitoids, E. mundus, Eretmocerus eremicus Rose & Zolnerowich and Encarsia formosa Gahan, and its influence in their susceptibility to insecticides, have been assessed. A total DNA extraction was performed on insects and universal bacterial primers were used to amplify the variable V4 region of the rRNA. A Next Generation sequencing (Illumina sequencing) was performed to identify the sequences of the bacterial genera present in the parasitic wasps. Once, the bacterial genera were identified, 16S rDNA gene of Actinobacteria were amplified from insects DNA extracts using the universal bacterial and actinobacterial primers, and the nested PCR products, were cloned to identify the Arthrobacter species. Three bacteria (A. aurescens Phillips, A. nicotinovarans Kodama, Yamamoto, Amano and Amichi and A. uratoxydans Stackebrandt, Fowler, Fiedler and Seiler), having the closest match with the Arthrobacter species present in the parasitic wasps, were obtained from the BCCMTM/LMG bacteria collection and its esterase activity was measured. Finally, antibiotic and residual contact tests were done to determine the influence of Arthrobacter species in the susceptibility of E. mundus to pesticides (abamectin). The results suggest that this bacterial genus can affect the toxicity of E. mundus to abamectin, which in turn supports the importance of the microbial community in natural enemies that it should be considered as a factor in risk assessment tests of pesticides.

Amblyseius largoensis in controlling red palm mite under semi-field conditions.

The objective of this work was to evaluate the efficiency of two Amblyseius largoensis (Acari:Phytoseiidae) populations in controlling Raoiella indica (Acari: Tenuipalpidae). The treatments were: release of A. largoensis from the island of La Réunion; release of A. largoensis from the state of Roraima, Brazil; and a control, without predator release. Initially, 20 predators were released per plant; three other releases were done at a rate of ten adults per plant, at 46, 135, and 156 days after the first release. The population densities were estimated every 20 days, during six months. Both A. largoensis populations evaluated are not sufficiently efficient to control the R. indica population.

Complete genomic sequence of a Rubus yellow net virus isolate and detection of genome-wide pararetrovirus-derived small RNAs

Rubus yellow net virus (RYNV) was cloned and sequenced from a red raspberry (Rubus idaeus L.) plant exhibiting symptoms of mosaic and mottling in the leaves. Its genomic sequence indicates that it is a distinct member of the genus Badnavirus, with 7932. bp and seven ORFs, the first three corresponding in size and location to the ORFs found in the type member Commelina yellow mottle virus. Bioinformatic analysis of the genomic sequence detected several features including nucleic acid binding motifs, multiple zinc finger-like sequences and domains associated with cellular signaling. Subsequent sequencing of the small RNAs (sRNAs) from RYNV-infected R. idaeus leaf tissue was used to determine any RYNV sequences targeted by RNA silencing and identified abundant virus-derived small RNAs (vsRNAs). The majority of the vsRNAs were 22-nt in length. We observed a highly uneven genome-wide distribution of vsRNAs with strong clustering to small defined regions distributed over both strands of the RYNV genome. Together, our data show that sequences of the aphid-transmitted pararetrovirus RYNV are targeted in red raspberry by the interfering RNA pathway, a predominant antiviral defense mechanism in plants. © 2013.

Effect of phenolic-rich plant materials on protein and lipid oxidation reactions

The antioxidant activity of natural plant materials rich in phenolic compounds is being widely investigated for protection of food products sensitive to oxidative reactions. In this thesis plant materials rich in phenolic compounds were studied as possible antioxidants to prevent protein and lipid oxidation reactions in different food matrixes such as pork meat patties and corn oil-in water emulsions. Loss of anthocyanins was also measured during oxidation in corn oil-in-water emulsions. In addition, the impact of plant phenolics on amino acid level was studied using tryptophan as a model compound to elucidate their role in preventing the formation of tryptophan oxidation products. A high-performance liquid chromatography (HPLC) method with ultraviolet and fluorescence detection (UV-FL) was developed that enabled fast investigation of formation of tryptophan derived oxidation products. Byproducts of oilseed processes such as rapeseed (Brassica rapa L.), camelina (Camelina sativa) and soy meal (Glycine max L.) as well as Scots pine bark (Pinus sylvestris) and several reference compounds were shown to act as antioxidants toward both protein and lipid oxidation in cooked pork meat patties. In meat, the antioxidant activity of camelina, rapeseed and soy meal were more pronounced when used in combination with a commercial rosemary extract (Rosmarinus officinalis). Berry phenolics such as black currant (Ribes nigrum) anthocyanins and raspberry (Rubus idaeus) ellagitannins showed potent antioxidant activity in corn oil-in-water emulsions toward lipid oxidation with and without β-lactoglobulin. The antioxidant effect was more pronounced in the presence of β-lactoglobulin. The berry phenolics also inhibited the oxidation of tryptophan and cysteine side chains of β-lactoglobulin. The results show that the amino acid side chains were oxidized prior the propagation of lipid oxidation, thereby inhibiting fatty acid scission. In addition, the concentration and color of black currant anthocyanins decreased during the oxidation. Oxidation of tryptophan was investigated in two different oxidation models with hydrogen peroxide (H2O2) and hexanal/FeCl2. Oxidation of tryptophan in both models resulted in oxidation products such as 3a-hydroxypyrroloindole-2-carboxylic acid, dioxindolylalanine, 5-hydroxy-tryptophan, kynurenine, N-formylkynurenine and β-oxindolylalanine. However, formation of tryptamine was only observed in tryptophan oxidized in the presence of H2O2. Pine bark phenolics, black currant anthocyanins, camelina meal phenolics as well as cranberry proanthocyanidins (Vaccinium oxycoccus) provided the best antioxidant effect toward tryptophan and its oxidation products when oxidized with H2O2. The tryptophan modifications formed upon hexanal/FeCl2 treatment were efficiently inhibited by camelina meal followed by rapeseed and soy meal. In contrast, phenolics from raspberry, black currant, and rowanberry (Sorbus aucuparia) acted as weak prooxidants. This thesis contributes to elucidating the effects of natural phenolic compounds as potential antioxidants in order to control and prevent protein and lipid oxidation reactions. Understanding the relationship between phenolic compounds and proteins as well as lipids could lead to the development of new, effective, and multifunctional antioxidant strategies that could be used in food, cosmetic and pharmaceutical applications.

The effect of temperature, photosynthetic photon flux density, and photoperiod on the vegetative growth and flowering of 'Autumn Bliss' raspberry

The effects of temperature, photosynthetic photon flux density (PPFD) and photoperiod on vegetative growth and flowering of the raspberry (Rubus idaeus L.) 'Autumn Bliss' were investigated. Increased temperature resulted in an increased rate of vegetative growth and a greater rate of progress to flowering. Optimum temperatures lay in the low to mid 20degreesC range. Above this the rate of plant development declined. Increased PPFD also advanced flowering. While photoperiod did not significantly affect the rate of vegetative growth, flowering occurred earliest at intermediate photoperiods and was delayed by extreme photoperiods. These responses suggest that there is potential for adjusting cropping times of raspberry grown under protection by manipulating the environment, especially temperature.

Diversity of phytoseiid mites (Acari: Mesostigmata: Phytoseiidae) in the Atlantic Forest of São Paulo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Gamasida (Arachnida: Acari) Edáficos de Piracicaba, Estado de São Paulo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)