Aflatoxins are carcinogenic mycotoxins most frequently associated with the filamentous fungus, Aspergillus flavus. These potent toxins pose serious health threats and their concentrations in foods are widely regulated. Maize, a critical staple of billions, is frequently contaminated with aflatoxins. Development of commercial maize hybrids with superior resistance to aflatoxin contamination has been sought for over 30 years without success. Analyses of native maize land races (MLRs) from Mexico revealed several accessions with significant resistance to both aflatoxin contamination and fungal reproduction. Physical barriers are important components of MLRs resistance. Traditional use of MLRs may reduce human exposure to aflatoxins. MLRs may contribute resistant genes of significant value in breeding for aflatoxin resistance. In Mexico, maize is produced from<10 to over 2,000 masl. Elevation had only minor influence on community compositions of aflatoxin-producing fungi associated with maize over three years in Sonora, Mexico. Most variation in community structure occurred between years. Dominant vegetative compatibility groups (VCGs) differed among years but were detected in similar frequencies across four agroecological zones. Multiple locations and multiple years must be sampled to obtain realistic assessments of the most successful VCGs. Elevated frequencies of VCG YV150 throughout Sonora during 2006 led to investigate genetic diversity within this VCG using microsatellite loci. The 2006 increases were attributed to rapid increase of a single clone. Examination of YV150 isolates collected over 20 years in the US and Mexico revealed several haplotypes and two genetically distinct populations, which were composed of isolates containing only a population specific mating-type idiomorph. Microsatellite loci in each population were in gametic equilibrium. Gene flow between isolates with different idiomorphs was not detected. VCG YV36, to which the biocontrol agent AF36 belongs, was found to be endemic to Mexico. Microsatellite loci revealed diversity within YV36 from Mexico, but all isolates harbored the single nucleotide polymorphism in the aflatoxin polyketide synthase gene, pksA, that confers atoxigenicity to AF36. Three YV36 isolates also had deletions in pksA suggesting continued degradation. Presence of endemic YV36 isolates in Mexico may facilitate rapid regulatory approval of AF36 for use in prevention of aflatoxin contamination of maize in Mexico.

No description available.

La famille des Phytoseiidae comprend des espèces d'intérêt agronomique dont certaines sont utilisées pour lutter contre des acariens ravageurs appartenant principalement aux familles des Tetranychidae et Eriophyidae. L'identification et la description des espèces de Phytoseiidae sont essentiellement basées sur des caractères morphologiques. Cependant, plusieurs problèmes d'identification sont rencontrés du fait : (1) de leur petite taille (faible nombre de caractères observables), (2) du manque de recul sur la valeur diagnostique de certains caractères dits discriminants, (3) et enfin du diagnostic fiable uniquement au stade femelle. Dans ce contexte, ce travail de thèse a pour objectif de déterminer dans quelle mesure l'approche moléculaire peut assister le diagnostic traditionnel. Pour cela, nous avons étudié l'étendue des variations intra- et interspécifiques (moléculaires et morphologiques) de sept espèces de Phytoseiidae d'intérêt agronomique, ceci pour comparer la capacité discriminante de quatre marqueurs moléculaires (12S ARNr, Cytb, COI et ITS). Dans ce travail, nous mettons en évidence l'utilité de ces marqueurs pour le diagnostic de tous les stades de développement et montrons l'importance de la forme du calyx de la spermathèque pour différencier deux espèces du genre Euseius, ce qui nous a permis de décrire une nouvelle espèce. Nous montrons également que les marqueurs mitochondriaux et nucléaires peuvent différencier des espèces morphologiquement proches. Cependant, il faut rester prudent quand aux conclusions tirées (espèces différentes ou populations différentes au sein d'une même espèce) sur la base des résultats obtenus avec les marqueurs mitochondriaux. Enfin, nous montrons que les règles de décision (valeurs seuils) sont difficiles à (1) établir avec ces mêmes marqueurs et (2) surtout à généraliser pour la différenciation des espèces appartenant aux deux sous-familles étudiées. Pour conclure, ce travail met en évidence de façon claire la nécessité d'une approche taxonomique intégrative, incluant un va-et-vient entre données moléculaires, morphologiques et écologiques, pour la garantie d'un diagnostic fiable des espèces de Phytoseiidae. / The family Phytoseiidae includes numerous species of huge interest for biological control of mite pests, essentially belonging to the families Tetranychidae and Eriophyidae. Identification and description of Phytoseiidae species are mainly based on morphological characters. However, taxonomists have to face several problems for species diagnostic because of : (1) their small size (low number of visible characters), (2) little information on the diagnostic value of some characters considered as discriminant, and at last (3) diagnostic only reliable at female stage. In this context, the present works aims to determine how molecular approaches can assist traditionnal diagnostic. For this, we studied the intra- and interspecific variations (morphological and molecular) of seven agronomic important species, in order to compare the ability of four molecular markers (12S rRNA, Cytb mtDNA, COI mtDNA et ITS) to discriminate between species. We first show that molecular markers could be of great help for differentiating species at all life stages. Moreover, we show the importance of the spermatheca calyx shape for separating two species of the genus Euseius, and a new specie was then described. We also emphasize that both mitochondrial and nuclear markers differentiate morphological close species. However, some cautions have to be paid on the conclusions developped (different species or different populations within a same species) when only mitochondrial markers are used. At last, we underline that decision rules (treshold cutt-off values) are difficult (1) to establish with these latter markers and also (2) to generalise for the accurate differentiation of the Phytoseiidae species belonging to the two sub-families considered. To conclude, this work clearly emphasizes the importance of integrative taxonomy, including an « up-and-down » analysis of molecular, morphological and ecological data, to secure the specific diagnostic among the family Phytoseiidae

M.Sc. / Please refer to full text to view abstract

Natural enemies provide critical population regulation of many pest species, though their effects are not commonly incorporated into agricultural management decisions. Conservation biological control is an important tool that can be implemented to minimize pest damage, but applying it requires appropriate understanding of pest and natural enemy relationships. Through experimental cotton field trials, I identified predator: prey ratios based on key arthropod predators as action thresholds of the whitefly pest Bemisia tabaci MEAM1 (Dinsdale et al. 2010; equivalent to Bemisia argentifolii Bellows et al. 1994 [Hemiptera: Aleyrodidae]), validated their efficacy, and promoted them to cotton pest managers. This dissertation begins with a multi-year field trial where whitefly and natural enemy populations were manipulated with a series of insecticidal treatments to identify key arthropod predators. The critical abundance of four key predators necessary to suppress whiteflies was estimated through predator: prey ratios. These ratios were refined for commercial pest management and developed to conform to the current whitefly IPM framework as a simple to use management-decision tool that would be readily adopted and used by pest managers. Predator: prey ratios were then validated in 1) a second field trial, 2) commercial fields in Arizona and northern Mexico and 3) historical field trials conducted from 1997-2010, where whitefly management decisions made with the standard threshold and ratios, were compared with the standard threshold alone. I found no difference in management outcomes when decisions were made with the standard threshold alone, or with predator: prey ratios in the field trial, but analysis of potential decisions on commercial farms and with historical trial data indicated that the majority of sprays could be delayed if control decisions incorporated ratio-based thresholds. Finally, an outreach program was developed and deployed to present ratios as decision-making tools for cotton pest managers that reduce uncertainty in control decisions and optimize spray outcomes. Pest managers indicated positive changes in knowledge and a gradual adoption of ratios for decision-making. The implementation of whitefly control decisions that incorporate predator: prey ratios may reduce pest managers' uncertainty in decision-making, as well as insecticide use and management costs.

Native to Asia, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) is an invasive non-native ladybird in Europe and North America, where it was widely introduced as a biological control agent of aphids and coccids. There were three main objectives of this study: firstly, to investigate the spread and distribution of H. axyridis in Europe from the start of the invasion process; secondly, to study its ecology in Great Britain; and thirdly, to examine its effects on native ladybirds in Britain. In Europe, collaborations with a network of scientists allowed the collation of H. axyridis occurrence data from across the continent. In Great Britain, a web-based public survey (www.harlequin-survey.org) was used, leading to the receipt of over 10,000 verified records of H. axyridis between 2004 and 2008. National land cover data enabled the habitats used by H. axyridis to be identified across Great Britain. These datasets were analysed in order to study the spread and ecology of H. axyridis in Europe and Britain. Fieldwork over a three year period (2006 to 2008) was carried out to examine changes in ladybird species assemblages during the invasion phase of H. axyridis in eastern England. Laboratory work was conducted to detect intraguild predation by H. axyridis, through PCR analyses of gut contents of field-collected ladybirds. In Europe, H. axyridis has spread since 2001 at the rate of approximately 200km yr-1. It has become established in at least 23 European countries. The established range extends from Norway in the north to southern France in the south, and from Ukraine in the east to Great Britain in the west. In the first five years of establishment H. axyridis spread north through Britain at the rate of 105km yr-1 and west at the rate of 145km yr-1. Evidence of the production of two generations per year was found, giving H. axyridis an advantage over most native ladybirds in Britain. Although H. axyridis was very common in urban habitats, it increasingly used semi-natural habitats. In addition, whilst the species was most common on deciduous trees, it was increasingly found on herbaceous plants. Aceraceae, Rosaceae and Malvaceae were the dominant plant families used by H. axyridis, especially for breeding. In eastern England H. axyridis increased from 0.1% to 40% of total ladybirds in three years, whilst native aphidophagous ladybirds declined from 84% to 41% of total ladybirds in the same period. Three species in particular experienced declines: Adalia 2-punctata, Coccinella 7-punctata and Propylea 14-punctata. Harmonia axyridis was the most abundant species by the end of the study. Detection of intraguild predation by one coccinellid on another, in the field in Europe, was shown for the first time using PCR techniques: A. 2-punctata DNA was detected in the gut of one of 112 field-collected H. axyridis. Harmonia axyridis has spread very quickly since 2001 and has become one of the most widely distributed coccinellids in Europe. Populations of native aphidophagous ladybirds were negatively affected by the arrival of H. axyridis, partly through intraguild predation.

Eastern and Carolina hemlock in the eastern United States are experiencing high mortality due to the invasive non-native hemlock woolly adelgid (HWA). The most promising means of control of HWA is the importation of natural enemies from the native range of HWA for classical biological control. Prior to release, natural enemies must be tested for suitability as a control agent, including the ability to locate the target prey. Coleopteran predators, including Scymnus coniferarum and Laricobius osakensis are under consideration as a means of biological control of HWA. Laricobius nigrinus was released in hemlock forests in 2003. It was recently discovered to hybridize with the native Laricobius rubidus. Behavioral responses of these predators to HWA and host tree foliage were observed using a 4-chambered olfactometer, and genetic analysis was used to differentiate responses of L. nigrinus, L. rubidus, and hybrids. In the olfactometer, insects are allowed to amble about the arena and respond to volatile cues from each treatment. Host foliage with and without HWA was tested, as were various comparisons of eastern versus western foliage, host versus non-host foliage, and foliage containing HWA and a congeneric feeding beetle. Olfactometer bioassays demonstrated that foliage from hosts where prey is commonly found is preferable to foliage where prey is seldom found, and that the presence of HWA-induced volatile cues is the strongest driver of behavior, and trumps the presence of a competitor. There is evidence in the study that supports the reliability-detectability phenomenon common in parasitoid biological control agents. Hybrid individuals were found to behave similarly to released L. nigrinus, although in some cases intermediate behavioral traits were evident, with respect to the parental species. This study and others support the continued need for strict testing of potential biological control agents prior to release, as well as a strong impetus for the inclusion and implementation of genetic analysis as a standard component of agent evaluation.

My master thesis deals with biology and morphology of the species Anaphes flavipes (Chalcidoidea: Mymaridae) and its potential use for biological control of leaf beetles. Particularly, the fitness of wasps was studied and a statistically significant difference in number of parasitize host eggs for mated females compared to non-mated. The effect of feeding parental population on the number of parasitized host eggs, the number of offspring in F1 generation or their sex ratio, was not proved. Study of host specificity of A. flavipes indicates preference for host eggs of species Oulema melanopus. Preference for heavier host eggs was not confirmed. The effect of host eggs weight on number of eggs wasps inside was not conclusive either. Length of parasitism of host eggs was consistent with earlier studies. The passive defense of host O. gallaecina against parasitoid was observed. Variability of the species depending on location, host, nutrition, etc. was determined using morphological measurements. The third part of the thesis is focused on population density of A. flavipes and its hosts in the localities of organic versus conventional farming in order to use the parasitoid for biological control. There was no significant difference in the size of pest and parasitoid populations between the types of agriculture....

Raoiella indica é uma severa praga de coqueiro, bananeira e plantas de importância paisagística. Os danos causados nas plantas hospedeiras iniciam-se pelo amarelecimento das folhas basais, seguida por necrose dos tecidos e secamento das folhas. Os objetivos desse trabalho foram: determinar a distribuição de R. indica e ácaros associados no centro-sul do Brasil; determinar os níveis populacionais de R. indica no estado de São Paulo; avaliar a sobrevivência de R. indica na ausência de alimento e diferentes níveis de umidade relativa; avaliar o potencial de ácaros fitoseídeos sobre R. indica; e avaliar o efeito de produtos químicos sobre esta praga. Raoiella indica foi encontrado no Distrito Federal e em 49 municípios de 9 estados, das quais, o mais meridional foi Iporã, estado do Paraná. Dos ácaros Mesostigmata associados a praga, Euseius citrifolius Denmark & Muma foi a espécie mais amplamente distribuída, sendo coletado em quase todos os estados. Os níveis populacionais de R. indica variaram de entre 0,2 a 2,5 ácaros/cm2 (46 a 599 ácaros por folíolo) em Marília e de 0,3 a 1,4 ácaros/cm2 (88,2 a 418 ácaros por folíolo) em Piracicaba. Euseius citrifolius foi o predador mais abundante em Marília, enquanto Amblyseius tamatavensis Blommers foi o predador mais abundante em Piracicaba. O tempo de sobrevivência de R. indica e T. urticae na ausência de alimento em diferentes níveis de umidade foi maior na fase adulta. Umidades abaixo de 47% diminuíram drasticamente o tempo de sobrevivência de todos os estágios. Das espécies de Phytoseiidae, A. tamatavensis mostrou melhor desempenho de predação e oviposição quando alimentados com ovos e ninfas de R. indica, como também se desenvolveu e reproduziu alimentando-se exclusivamente desta praga. Dos produtos testados, o fenpiroximato causou mortalidade de 100% em R. indica. Dos produtos utilizados sobre os predadores, o fenpiroximato foi o mais tóxico para os fitoseídeos, sobretudo em Iphiseiodes zuluagai Denmark & Muma. Os resultados desse estudo sugerem que o estabelecimento de R. indica além do sul do Paraná é possível em microclimas favoráveis ao seu desenvolvimento. Além disso, os estudos indicaram que em São Paulo os altos níveis de precipitação no verão e as baixas temperaturas durante o inverno afetam negativamente os níveis de R. indica. A possibilidade de uso de A. tamatevensis como agente de controle de R. indica precisa ser melhor estudada em condições de campo. / Raoiella indica is a severe pest of coconut, banana trees, and floristic plants. The damage caused to the host plants begins by basal leaves yellowing, followed by tissue necrosis and drying of the leaves. The objectives of this study were: to determine the distribution of R. indica and associated mites in the south-central Brazil; determine population levels of R. indica in São Paulo state; assess the survival of R. indica in the absence of food and different levels of relative humidity; assess the potential of phytoseiids mites on R. indica; and to evaluate the effect of chemicals on the pest. Raoiella indica has been found in the Federal District and in 49 municipalities in 9 States, of which the southernmost was Iporã, Paraná state. Among the mesostigmatid mites associated with this pest, Euseius citrifolius Denmark & Muma was the most widely distributed species occurring in almost all states considered in this study. In Sao Paulo, levels of R. indica varied of between 0.2 to 2.5 dust mites/cm2 (46 to 599 mites per leaflet) in Marília and 0.3 to 1.4 mites/cm2 (88.2 to 418 mites per leaflet) in Piracicaba. Euseius citrifolius was the most abundant predator in Marília. In Piracicaba, Amblyseius tamatavensis Blommers was more abundant. The survival time of R. indica and T. urticae in the absence of food in different levels of moisture was greater in the adult stage. Humidities below 47% decreased drastically the time of survival of all stages. Amblyseius tamatavensis showed better performance of oviposition and predation when fed with eggs and nymphs of R. indica, and developed and reproduced feeding exclusively on this pest. Of the products tested, fenpiroximate caused 100% mortality in R. indica. All products had efficiency above 80%. Fenpiroximate was most toxic for phytoseiids, especially of Iphiseiodes zuluagai Denmark & Muma. The results of this study suggest that the establishment of R. indica beyond southern Paraná is possible in microclimates favorable to its development. In addition, the studies confirm that in São Paulo the high levels of precipitation in summer and low temperatures during winter negatively affect the levels of R. indica. The possibility of using A. tamatavensis as a biological control agent of R. indica requires further evaluation.

Alimentos produzidos de forma mais sustentável têm ganhado espaço e o controle biológico vem assumindo importância cada vez maior em programas de manejo integrado de pragas. Dentre os principais agentes de controle biológico estão os ácaros predadores pertencentes a família Phytoseiidae. Amblyseius tamatavensis Blommers é um ácaro predador pertencente a esta família, amplamente distribuído no Brasil e no mundo. Este predador apresentou resultados promissores em relação ao possível uso para o controle de Bemisia tabaci (Gennadius). Alguns estudos mostraram diferenças biológicas marcantes entre as populações de fitoseídeos. Portanto, espera-se que tais diferenças também possam ocorrer entre as populações brasileiras de A. tamatavensis. Por isso, 14 populações de diferentes regiões do Brasil (dos estados de Alagoas, Goiás, Minas Gerais e São Paulo) foram comparadas em relação ao potencial de predação e oviposição quando alimentadas com ovos de B. tabaci biótipo B. A população coletada em Olhos d\'Água das Flores (Alagoas) apresentou as maiores taxas de predação diária (7,9 ovos / fêmea) e oviposição (1,2 ovo / fêmea) e uma variabilidade entre os indivíduos da população que possibilitou um processo de seleção. O objetivo deste trabalho foi selecionar em laboratório uma linhagem de A. tamatavensis com maior potencial de predação e oviposição, sendo assim mais favorável para uso prático no controle de B. tabaci. Na seleção para predação, após seis gerações, foi observado um ganho genético de 1,1 ovo/fêmea/dia (aproximadamente 15% de aumento em relação à população) na taxa de predação, demonstrando resultado promissor para o melhoramento. Entretanto, o ganho genético foi praticamente nulo quando a população selecionada para predação foi submetida ao processo de seleção para oviposição, e também foi observado um baixo diferencial de seleção, mostrando um baixo potencial para a seleção das melhores fêmeas em relação a este parâmetro. Em função dos resultados promissores em relação ao aumento da eficiência da população de A. tamatavensis no controle de B. tabaci e das tabelas de vida de fertilidade não mostrarem custos adaptativos importantes, testes em plantas de pimentão mantidas em um telado foram realizados para avaliar a real eficiência desta linhagem. Foram comparadas diferentes populações, selecionadas e não selecionadas em laboratório. Os melhores resultados foram observados com a população que passou pelo processo de seleção, que teve uma eficiência maior quando comparada com as outras populações. Essa população apresentou uma redução nos imaturos de B. tabaci que variou de 82% a 90% após 14 dias da liberação dos predadores. Os resultados obtidos nas plantas de pimentão sugerem que, assim como observado nas tabelas de vida realizadas no capítulo anterior, não houve um custo adaptativo significativo na população que passou pelo processo de seleção. Estudos complementares devem ser realizados, para avaliar se essas populações estudadas mantém o mesmo comportamento em cultivos no campo. / More attention has been given recently to sustainable food production, and biological control has become increasingly important in integrated pest management programs. Predatory mites of the family Phytoseiidae are among the main biological control agents. Amblyseius tamatavensis Blommers is a predatory mite belonging to this family, widely distributed in Brazil and worldwide. This predator presented promising results regarding the possible use for the control of Bemisia tabaci (Gennadius). Some studies have shown marked biological differences among phytoseiid populations. It is expected that such differences may also occur among Brazilian populations of A. tamatavensis. Therefore, 14 populations of different regions of Brazil (from the states of Alagoas, Goiás, Minas Gerais and São Paulo) were compared in relation to predation and oviposition potential when fed with B. tabaci biotype B. The population collected in Olhos d\'Água das Flores (Alagoas) presented the highest rates of daily predation (7.9 eggs / female) and oviposition (1.2 egg / female) and sufficient variability among individuals of the population to enable a selection process. The objective of this work was to select in the laboratory a strain of A. tamatavensis with higher predation and oviposition potential, being thus more favorable for practical use in the control of B. tabaci. In the selection for predation, after six generations, a genetic gain of 1.1 eggs / female / day (approximately 15% increase in relation to the population) in predation rate was observed, showing promising result for breeding. However, the genetic gain was practically null when the population selected for predation was submitted to the selection process for oviposition, and a low selection differential was also observed, showing a low potential for the selection of the best females in relation to this parameter. As a result of the promising results in increasing the efficiency of the A. tamatavensis population in the control of B. tabaci and because the fertility life tables did not show important adaptive costs, tests on pepper plants were carried in a screen-house to evaluate the real efficiency of this strain. Different populations were compared, selected and not selected in the laboratory. The best results were observed with the population that passed by the selection process, which had a greater efficiency when compared with the other populations. This population showed a reduction in the immatures of B. tabaci that ranged from 82% to 90% after 14 days of predator release. The results obtained on the pepper plants suggest that, as observed in the life tables performed previously, there was no significant adaptive cost in the population that passed by selection process. Complementary studies should be carried out to evaluate whether these studied populations maintain the same behavior in field crops.