Augmentation of the assassin bug Pristhesancus plagipennis (Walker) (Hemiptera : Reduviidae) as a biological control agent for Helicoverpa spp. in cotton

Third-instar nymphs of the Australian assassin bug, Pristhesancus plagipennis (Walker), were released into cotton plots at two release densities and two crop growth stages to test their biological control potential. Release rates of 2 and 5 nymphs per metre row resulted in field populations of 0.51 and 1.38 nymphs per metre row, respectively, indicating that over 70% of nymphs died or emigrated within two weeks of release. Effective release rates of 1.38 nymphs per metre row reduced the number of Helicoverpa spp. larvae in the plots for a 7-week period. Crop yields were significantly greater in the plots to which P. plagipennis nymphs were released, with the effective release rate of 1.38 nymphs per metre row providing equivalent yields as insecticide treated plots. The data suggest that P. plagipennis has the capacity to reduce Helicoverpa spp. larvae densities in cotton crops when augmented through inundative release.

Haematophagy and cleptohaematophagy of Clerada apicicornis (Hemiptera: Lygaeidae), a potential biological control agent of Rhodnius prolixus (Hemiptera: Reduviidae)

Because of its ability to prey on Triatominae in rural houses, Clerada apicicornis has been suggested as a potential biological control agent of Rhodnius prolixus. It has also been suggested as a potential vector of mammalian trypanosomes such as Trypanosoma cruzi, because of its ability to take blood directly from mammals. To help resolve these conflicting ideas, we assessed the haematophagic behaviour of C. apicicornis by carrying out feeding trials on laboratory animals. Cleptohaematophagic behaviour was also assessed by allowing C. apicicornis to feed on R. prolixus previously engorged with avian blood. The low proportion of blood meals taken directly from laboratory animals indicates a facultative haematophagy in this species, whereas a greater proportion of nymphs and adults were able to obtain vertebrate blood by predation on engorged R. prolixus. The results suggest that C. apicicornis is unlikely to be effective as a biological control agent, but is also unlikely to have a significant role in the transmission of vertebrate pathogens.

Regulation of mucoidy in the biological control agent Pseudomonas fluorescens CHA0 and construction of an exopolysaccharide-overproducing variant

Pseudomonas fluorescens CHA0, an effective biological control agent of soilborne plant diseases, is naturally non-mucoid. We have isolated a highly mucoid Tn5 insertion mutant of strain CHA0. The mucoid phenotype was found to be due to the overproduction of exopolysaccharide (EPS), as a result of a mutation in the mucA gene. The wild-type mucA gene was cloned by a two-step, Tn5-dependent cloning procedure previously described and the deduced amino acid sequence showed 71% identity with MucA of P. aeruginosa, a negative regulator of the alternative sigma factor AlgU (=s22, sE). As in P. aeruginosa, mucA is preceded by the algU gene encoding s22 (91% identity at the amino acid sequence level). A mucA in-frame deletion mutant of CHA0 overproduced EPS and formed mucoid colonies, whereas an algU in-frame deletion mutant showed a non-mucoid phenotype. Pyoluteorin, an antibiotic produced by P. fluorescens, was found to be entrapped in EPS of a mucoid mutant. In natural soil, mucoidy negatively affected survival of the bacteria, suggesting that under these conditions the potential to produce abundant EPS does not confer a selective advantage on the bacteria.

Use of grass carp (Ctenopharyngodon idella) as a biological control agent for submerged aquatic macrophytes

This study aimed to evaluate feed preference and control efficacy of grass carp (Ctenopharyngodon idella) on the aquatic macrophytes Ceratophyllum demersum, Egeria densa and Egeria najas. An experiment was carried out at mesocosms conditions with 2,000 liters capacity and water residence time of 2.8 days. C. demersum, E. densa e E. najas biomasses were offered individually with sixty g and coupled in similar quantities of 30 g of each species, evaluated during 81 days, envolving 6 treatments. (1 - C. demersum, 2 - E. najas, 3 -E. densa, 4 - C. demersum + E. najas, 5 - C. demersum + E. densa and 6 - E. najas + E. densa). When offered individually, E. najas and C. demersum presented the same predation rate by grass carp, which was higher than E. densa predation rate. When plants were tested in pairs, the order of feed preference was C. demersum > E. najas > E. densa. E. najas and C. demersum percentage control ranged from 73 to 83%. No relation between biomass consumption and grass carp body weight gain was observed, probably due to differences in nutritional quality among macrophyte species according to fish necessities. Therefore, it is concluded that the use of grass carp is one excellent technique to control submersed macrophytes in Brazil.

Characterization and mechanism of action of the biological control agent Pantoea agglomerans EPS125

La soca EPS125 ha mostrat ser un efectiu agent de control biològic de diferents patògens fúngics de postcollita en diferents fruits. Degut a la seva elevada eficàcia, es va plantejar desenvolupar aquesta soca comercialment i per aquest motiu en el present treball es plantejà complementar la informació necessària pel seu registre. D'acord amb els resultats obtinguts mitjançant proves fenotípiques i genotípiques, la soca EPS125 queda inclosa dins l'espècie Pantoea agglomerans (Enterobacter agglomerans-Erwinia herbicola). En relació a la utilització de fonts de carboni, en el perfil i contingut d'àcids grassos cel·lulars i en el polimorfisme en la longitud dels fragments de macrorestricció genòmica (MRFLP), la soca EPS125 mostrà trets característics que la diferencien d'altres soques. Els dos marcadors moleculars (125.2 i 125.3) específics per la soca EPS125 dissenyats en el present treball mostraren ser semiespecífics per la seva detecció mitjançant la tècnica PCR i Real Time PCR. Quedant pendent l'anàlisi d'especificitat de l'ús combinat dels dos marcadors moleculars en una reacció PCR multiplex. P. agglomerans EPS125 ha mostrat ser molt efectiva en el control de Penicillium expansum en poma amb una dosi efectiva mitjana de 2.7x105 a 7x105 ufc/ml, i una ratio de 25-101 cèl·lules de la soca EPS125 per inactivar una espora del patogen segons el model de saturació hiperbòlica. Segons les aproximacions fenotípiques i estudis genotípics realitzats, sembla que els mecanismes de biocontrol utilitzats per la soca EPS125 contra P. expansum en poma estan directament relacionats amb la capacitat de formació de biofilm per aquesta soca.

Development of molecular monitoring methods and assessment of the environmental fate of the biological control agent of fire blight Pseudomonas fluorescens EPS62e

Pseudomonas fluorescens EPS62e es va seleccionar com a agent de biocontrol del foc bacterià per la seva eficàcia en el control de Erwinia amylovora. En aquest treball es van desenvolupar mètodes de traçabilitat que van permetre la seva detecció específica i quantificació. Mitjançant les tècniques RAPD i U-PCR es van obtenir fragments d'amplificació diferencial per EPS62e que es van seqüenciar i caracteritzar com marcadors SCAR per dissenyar una PCR en temps real. La PCR a temps real es va utilitzar simultàniament amb mètodes microbiològics per estudiar l'adaptabilitat epifítica de EPS62e en pomera i perera. L'ús combinat de mètodes microbiològics i moleculars va permetre la identificació de tres estats fisiològics de EPS62e: la colonització activa, l'entrada en un estat de viable però no cultivable, i la mort cel·lular. Aquest treball mostra que EPS62e està ben adaptada a la colonització de flors a camp, encoratjant la seva utilització dins d'una estratègia de control biològic contra el foc bacterià.

Host specificity of Metriona elatior, a potential biological control agent of tropical soda apple, Solanum viarum, in the USA

The leaf beetle Metriona elatior from Brazil-Argentina was screened in the Florida (USA) State quarantine facility as a potential biological control agent of tropical soda apple, Solanum viarum, a recently arrived weed species. Multiple-choice host-specificity tests were conducted in small cages (60 cm x 60 cm x 60 cm) using 95 plant species in 29 families. Adults fed heavily on the main target weed (S. viarum), and on turkeyberry, Solanum torvum (noxious weed of Asiatic origin); fed moderately on red soda apple, Solanum capsicoides (weed of South American origin), and eggplant, Solanum melongena (economic crop); and fed lightly on aquatic soda apple, Solanum tampicense (weed of Mexican-Caribbean-Central American origin), and on silverleaf nightshade, Solanum elaeagnifolium (native weed widely distributed). M. elatior adults laid 84 to 97% of their egg masses on S. viarum, and 3 to 16% on S. melongena. Non-choice host-specificity tests were also conducted in quarantine in which M. elatior adults and neonate larvae were exposed to 17 and 19 plant species, respectively. Tests with the neonates indicate that this insect was able to complete its development on S. viarum, S. torvum, S. melongena, and S. capsicoides. Although some adult feeding and oviposition occurred on S. melongena in quarantine on potted plants in small cages, no feeding or oviposition by M. elatior was observed in field experiments conducted in Brazil. Surveys in unsprayed S. melongena fields in Argentina and Brazil indicated that M. elatior is not a pest of S. melongena in South America. The evidence obtained from the South-American field surveys, Brazil open-field experiments, and Florida quarantine host specificity tests indicate that M. elatior causes significant feeding damage to S. viarum, and does not represent a threat to S. melongena crops in the USA. Therefore an application for permission to release M. elatior against S. viarum in the USA was submitted in October 1998.

Potential of Acarophenax lacunatus (Prostigmata: Acarophenacidae) as a biological control agent of Rhyzopertha dominica (Coleoptera: Bostrichidae)

A recent report of the parasitic mite species Acarophenax lacunatus (Cross and Krantz) (Prostigmata: Acarophenacidae) attacking populations of Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) led to the present investigation. Maximum female size and average number of progeny per female mite were assessed at eight different temperatures (ranging from 20 to 41°C) and 60% r.h. using R. dominica as the host. The ability of the mite species to suppress eggs, first instar larvae, and adults of R. dominica was assessed at 30°C and 60% r.h. The largest female sizes of the mite and progeny numbers were obtained around 30°C (259 μm and 17 offspring/female respectively) with minimum values obtained at the most extreme temperatures used in this study. Mite densities of at least four individuals per 500 ml jar containing 50 adults of R. dominica, resulted in almost complete suppression of eggs, first instar larvae, and adults of the host species after 45 days. This same range of mite densities led to reductions of wheat weight losses of 15 and 25% after 45 and 60 days after infestation respectively. Acarophenax lacunatus shows good potential as a biological control agent of R. dominica.

Saccharomyces cerevisiae: a novel and efficient biological control agent for Colletotrichum acutatum during pre-harvest

In this study, we evaluated the efficiency of six isolates of Saccharomyces cerevisiae in controlling Colletotrichum acutatum, the causal agent of postbloom fruit drop that occur in pre-harvest citrus. We analyzed the mechanisms of action involved in biological control such as: production of antifungal compounds, nutrient competition, detection of killer activity, and production of hydrolytic enzymes of the isolates of S. cerevisiae on C. acutatum and their efficiency in controlling postbloom fruit drop on detached citrus flowers. Our results showed that all six S. cerevisiae isolates produced antifungal compounds, competed for nutrients, inhibited pathogen germination, and produced killer activity and hydrolytic enzymes when in contact with the fungus wall. The isolates were able to control the disease when detached flowers were artificially inoculated, both preventively and curatively. In this work we identified a novel potential biological control agent for C acutatum during pre-harvest. This is the first report of yeast efficiency for the biocontrol of postbloom fruit drop, which represents an important contribution to the field of biocontrol of diseases affecting citrus populations worldwide. (C) 2015 Elsevier GmbH. All rights reserved.

Use of grass carp (ctenopharyngodon idella) as a biological control agent for submerged aquatic macrophytes

This study aimed to evaluate feed preference and control efficacy of grass carp (Ctenopharyngodon idella) on the aquatic macrophytes Ceratophyllum demersum, Egeria densa and Egeria najas. An experiment was carried out at mesocosms conditions with 2,000 liters capacity and water residence time of 2.8 days. C. demersum, E. densa e E. najas biomasses were offered individually with sixty g and coupled in similar quantities of 30 g of each species, evaluated during 81 days, envolving 6 treatments. (1 - C. demersum, 2 - E. najas, 3 - E. densa, 4 - C. demersum + E. najas, 5 - C. demersum + E. densa and 6 - E. najas + E. densa). When offered individually, E. najas and C. demersum presented the same predation rate by grass carp, which was higher than E. densa predation rate. When plants were tested in pairs, the order of feed preference was C. demersum > E. najas > E. densa. E. najas and C. demersum percentage control ranged from 73 to 83%. No relation between biomass consumption and grass carp body weight gain was observed, probably due to differences in nutritional quality among macrophyte species according to fish necessities. Therefore, it is concluded that the use of grass carp is one excellent technique to control submersed macrophytes in Brazil.

Biological Characteristics and Thermal Requirements of the Biological Control Agent Telenomus remus (Hymenoptera: Platygastridae) Reared on Eggs of Different Species of the Genus Spodoptera (Lepidoptera: Noctuidae)

The biological characteristics of Telenomus remus Nixon, 1937 (Hymenoptera: Platygastridae) on eggs of Spodoptera albula (Walker, 1857); S. cosmioides Walker 1858, S. eridania (Cramer, 1782); and S. frugiperda (Smith, 1797) (Lepidoptera: Noctuidae) were evaluated under different temperatures (19, 22, 25, 28, 31, and 34 degrees C +/- 1 degrees C). The duration of the T remus egg-to-adult period on eggs of all four Spodoptera species and the longevity of adults of T. remus were both inversely proportional to the increase in temperature. Parasitoid emergence was higher than 80% at temperatures from 19 to 28 degrees C when the parasitoid was reared on eggs of S. eridania and S. frugiperda. Differently, when the parasitoid was reared on eggs of S. albula and S. cosmioides, T. remus emergence at rates of 80% or higher just occurred from 22 to 25 degrees C and at 22 degrees C, respectively. At 34 degrees C, this parameter was lower than 30% for T reams reared in all hosts. The sex ratio was 64-86% females, except for T. remus in S. cosmioides eggs at 34 C, in which temperature it was 39%. The estimated thermal requirements of T. remus, for the thermal constant (K) and the base temperature (T(base)), were: 125.39 DD and 15.139 degrees C; 125.56 DD and 14.912 degrees C; 142.98 DD and 14.197 degrees C; and 149.16 DD and 13.846 degrees C, for S. cosmioides, S. frugiperda, S. albula, and S. eridania, respectively. In general, T. remus showed good parasitism potential on all the hosts, although eggs of S. frugiperda, S. eridania, and S. albula proved to be the most suitable for mass rearing of T reams in the laboratory. Eggs of S. cosmioides are less suitable because of the lower parasitoid emergence observed at most of the temperatures with exception of 22 degrees C.

Aureobasidium pullulans as biological control agent: modes of action

The postharvest phase has been considered an environment very suitable for successful application of biological control agents (BCAs). However, the tri-interaction between fungal pathogen, host (fruit) and antagonist is influenced by several parameters such as temperature, oxidative stresses, oxygen composition, water activity, etc. that could be determining for the success of biocontrol. Knowledge of the modes of action of BCAs is essential in order to enhance their viability and increase their potentialities in disease control. The thesis focused on the possibility to explain the modes of action of a biological control agent (BCA): Aureobasidium pullulans, in particular the strains L1 and L8, control effective against fruit postharvest fungal pathogen. In particular in this work were studied the different modes of action of BCA, such as: i) the ability to produce volatile organic compounds (VOCs), identified by SPME- gas chromatography-mass spectrometry (GC-MS) and tested by in vitro and in vivo assays against Penicillium spp., Botrytis cinerea, Colletotrichum acutatum; ii) the ability to produce lytic enzymes (exo and endo chitinase and β-1,3-glucanase) tested against Monilinia laxa, causal agent of brown rot of stone fruits. L1 and L8 lytic enzymes were also evaluated through their relative genes by molecular tools; iii) the competition for space and nutrients, such as sugars (sucrose, glucose and fructose) and iron; the latter induced the production of siderophores, molecules with high affinity for iron chelation. A molecular investigation was carried out to better understand the gene regulation strictly correlated to the production of these chelating molucules. The competition for space against M. laxa was verified by electron microscopy techniques; iv) a depth bibliographical analysis on BCAs mechanisms of action and their possible combination with physical and chemical treatments was conducted.

Beauveria bassiana as a biological control agent of the corn rootworm : final report /

"Project: 88/142."

Ecological genetics of Melaleuca quinquenervia (Myrtaceae): Population variation in Florida and its influence on performance of the biological control agent Oxyops vitiosa (Coleoptera: Curculionidae)

Melaleuca quinquenervia (Cav.) Blake (Myrtaceae) was imported into Florida from Australia over a century ago as a landscape plant. A favorable climate and periodic wildfires helped M. quinquenervia thrive; it now occupies about 200,000 hectares in southern Florida. A biological control (i.e., biocontrol) program against M. quinquenervia has been initiated, but not all biocontrol releases are successful. Some scientists have argued that poor biocontrol agent success may relate to genetic differences among populations of invasive weeds. I tested this premise by determining (1) the number and origins of M. quinquenervia introductions into Florida, (2) whether multiple introduction events resulted in the partitioning of Florida's M. quinquenervia populations into discrete biotypes, and (3) whether Oxyops vitiosa, an Australia snout beetle imported to control this weed, might discriminate among putative M. quinquenervia biotypes. Careful scrutiny of early horticultural catalogs and USDA plant introduction records suggested at least six distinct introduction events. Allozyme analyses indicated that the pattern of these introductions, and the subsequent redistribution of progeny, has resulted in geographic structuring of the populations in southern Florida. For example, trees on Florida's Gulf Coast had a greater effective number of alleles and exhibited greater heterozygosity than trees on the Atlantic Coast. Essential oil yields from M. quinquenervia leaves followed a similar trend; Gulf Coast trees yielded nearly twice as much oil as Atlantic Coast trees when both were grown in a common garden. These differences were partially explained by the predominance of a chemical phenotype (chemotype) very rich in the sesquiterpene (E)-nerolidol in M. quinquenervia trees from the Gulf Coast, but rich in a mixture of the monoterpene 1,8-cineole and the sesquiterpene viridiflorol in trees from the Atlantic Coast. Performance of O. vitiosa differed dramatically in laboratory studies depending on the chemotype of the foliage they were fed. Larval survivorship was four-fold greater on the (E)-nerolidol chemotype. Growth was also greater, with adult O. vitiosa gaining nearly 50% more biomass on the (E)-nerolidol plants than on the second chemotype. The results of this study thus confirmed the premise that plant genotype can affect the population dynamics of insects released as weed biocontrols. ^