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Assessing risks of releasing exotic biological control agents of arthropod pests.

More than 5000 introductions of about 2000 species of exotic arthropod agents for control of arthropod pests in 196 countries or islands during the past 120 years rarely have resulted in negative environmental effects. Yet, risks of environmental effects caused by releases of exotics are of growing concern. Twenty countries have implemented regulations for release of biological control agents. Soon, the International Standard for Phytosanitary Measures (ISPM3) will become the standard for all biological control introductions worldwide, but this standard does not provide methods by which to assess environmental risks. This review summarizes documented nontarget effects and discusses the development and application of comprehensive and quick-scan environmental risk assessment methods.

Animals↗

Biological control agents for white grubs (Coleoptera: Scarabaeidae) in anticipation of the establishment of the Japanese beetle in California.

We tested biological control agents for the control of 3rd-instar scarab turfgrass pests, both for the masked chafer Cyclocephala hirta LeConte and the Japanese beetle, Popillia japonica Newman. The former species is endemic in California whereas the latter, although not yet established, constitutes a permanent serious threat to agriculture and horticulture in California. We conducted experiments using C. hirta in California and P. japonica in New Jersey. A field trial conducted in 2 different California turfgrass sites compared the field persistence in the absence of hosts of Bacillus thuringiensis Berliner subspecies japonensis Buibui strain, the milky disease bacterium, Paenibacillus (=Bacillus) popilliae (Dutky), and the entomopathogenic nematodes Steinernema kushidai Mamiya and Heterorhabditis bacteriophora Poinar to that of the organophosphate diazinon. Soil samples taken 0-70 d after applications were bio-assayed with P. japonica. Only diazinon and the entomopathogenic nematode S. kushidai caused substantial mortality and S. kushidai activity persisted significantly longer than diazinon activity. In greenhouse experiments, combinations of entomopathogenic nematode species usually resulted in additive mortality of scarab larvae. Combinations of S. kushidai and diazinon also resulted in additive mortality. In field trials, the efficacy of H. bacteriophora and especially S. kushidai and S. glaseri, was comparable to that of diazinon over 14-18 d. However, it is likely that at least S. kushidai would have outperformed diazinon over an extended period because of its longer persistence and potential for recycling in the hosts. S. kushidai, should it become commercially available, deserves further examination as an alternative to chemical white grub control especially as a highly compatible component of sustainable turfgrass management.

Animals↗

Investigations on bacteria as a potential biological control agent of summer chafer, Amphimallon solstitiale L. (Coleoptera: Scarabaeidae).

Studying the bacteria of hazardous insects allows the opportunity to find potentially better biological control agents. Therefore, in this study, bacteria from summer chafer (Amphimallon solstitiale L., Coleoptera: Scarabaeidae) we isolated and identified the insecticidal effects of bacteria isolated from A. solstitiale and Melolontha melolontha L. (common cockchafer, Coleoptera: Scarabaeidae) and the mixtures of these bacterial isolates were investigated on A. solstitiale larvae. Crystals from Bacillus sp. isolated from M. melolontha were also purified, and tested against the second and third-stage larvae of A. solstitiale. The bacterial isolates of A. solstitiale were identified as Pseudomonas sp., Pseudomonas sp., Bacillus cereus and Micrococcus luteus, based on their morphology, spore formation, nutritional features, and physiological and biochemical characteristics. The insecticidal effects of the bacterial isolates determined on the larvae of A. solstitiale were 90% with B. cereus isolated from A. solstitiale, and 75% with B. cereus, B. sphaericus and B. thuringiensis isolated from M. melolontha within ten days. The highest insecticidal effects of the mixed infections on the larvae of A. solstitiale were 100% both with B. cereus+B. sphaericus and with B. cereus+B. thuringiensis. In the crystal protein bioassays, the highest insecticidal effect was 65% with crystals of B. thuringiensis and B. sphaericus isolated from M. melolontha within seven days. Finally, our results showed that the mixed infections could be utilized as microbial control agents, as they have a 100% insecticidal effect on the larvae of A. solstitiale.

Animals↗

[The role of Marisa cornuarietis as a biological control agent and its economic and epidemiological implications].

It was determined that M. cornuarietis, a mollusk which has been used as agent for the biological control of the schistosomiasis hosts, may be a plague for rice fields. Each mollusk can consume 0.3 g of this plant in 24 hours, accounting for the destruction of 0.015 m2 of a rice field. On the other hand, it was observed that B. glabrata shows preference for the consumption of M. cornuarietis faeces. This fact favors the vector's growth and reproduction rate and at the same time decreases its mortality.

Animals↗

In vivo rearing of Thripinema nicklewoodi (Tylenchida: Allantonematidae) and prospects as a biological control agent of Frankliniella occidentalis (Thysanoptera: Thripidae).

Methods are described for the in vivo production of the nematode Thripinema nicklewoodi (Siddiqi), an obligate parasite and potential biological control agent of western flower thrips Frankliniella occidentalis (Pergande). Nematode infection is not lethal but causes sterilization of adult female hosts. Both fertilization and horizontal transmission of T. nicklewoodi is achieved in 1.5-ml microcentrifuge tubes (infection arenas), in the presence of 100% humidity, a temporary food source and preferably a damp substrate. Following exposure to infection arenas, F. occidentalis are reared on excised bean leaves Phaseolus vulgaris (L.) in polypropylene containers for 2 wk at 25 degrees C to allow the reproduction and development of a single generation of nematodes within infected hosts's abdominal cavity. To identify infected hosts after this incubation period, thrips are isolated in microcentrifuge tubes and monitored for free-living nematodes being released along with frass. Infected thrips are reintroduced back into infection arenas to inoculate further thrips to maintain the culture. We documented the output of the rearing procedure using a standard method and following simple manipulation of several individual parameters of the infection technique. The standard method was the most efficient, and resulted in an increased (output/input) ratio of infected thrips of approximately 2; i.e., the number of infected thrips approximately doubles each generation. Monitoring infected thrips revealed that nematodes were first released between 12-14 d postinfection and for an average of 7.9 d at 25 degrees C; highlighting the potential to reuse infective thrips between infection arenas. The possibility of using T. nicklewoodi as an inoculative agent against F. occidentalis infesting floricultural crops is discussed.

Animals↗

Characterization of a Genomic Region Required for Production of the Antibiotic Pyoluteorin by the Biological Control Agent Pseudomonas fluorescens Pf-5.

A 21-kb region required for the biosynthesis of the polyketide antibiotic pyoluteorin by the biological control agent Pseudomonas fluorescens Pf-5 was identified and cloned. Seven previously isolated mutants deficient in pyoluteorin production (Plt(sup-)) had Tn5 insertions spanning the 21-kb region. Sequences flanking Tn5 inserts were cloned from genomic DNA of three Plt(sup-) mutants and used as probes to identify wild-type alleles of the plt loci from a genomic library of Pf-5. Five cosmids containing overlapping regions of genomic DNA hybridized to one or more of the probes. One cosmid, pJEL1938, contained the entire 21-kb region and, when introduced into a Plt(sup-) mutant, partially restored pyoluteorin production. To study the expression of the genes required for pyoluteorin biosynthesis, the transposon Tn3-nice, which contains a promoterless ice nucleation gene (inaZ) and a type I neomycin phosphotransferase gene, was introduced into the genomic plt region of Pf-5. Carbon sources that influenced pyoluteorin production by Pf-5 had parallel effects on ice nucleation activity of Pf-5 containing a genomic plt::Tn3-nice fusion, indicating that inaZ was transcribed from a promoter of the plt region. Cells of Pf-5 containing a genomic plt::Tn3-nice fusion expressed ice nucleation activity on cotton and cucumber seeds planted in field soil. The expression of plt genes by Pf-5 in the cucumber spermosphere was delayed in comparison with expression in the cotton spermosphere. This study demonstrates that genes required for pyoluteorin production were expressed in situ by the biological control bacterium.

Journal Article↗

Mesocyclops brevisetosus (Cyclopoida: Cyclopoidae) as a potential biological control agent against mosquito larvae in Indonesia.

Mesocyclops brevisetosus Dussart and Sarnita, an indigenous copepod in Indonesia, was evaluated as a potentially useful biological control agent against larval Aedes (Stegomyia). In the laboratory, female M. brevisetosus were found to readily consume mosquito larvae of species from 3 genera (Aedes aegypti, Culex quinquefaciatus, and Anopheles farauti). Mesocyclops brevisetosus preferentially fed on larvae of Ae. aegypti (mean 95% consumed), followed by Cx. quinquefasciatus (71%) and An. farauti (54%) during a 24-h period. An individual copepod consumed up to 43 1st and 2nd instars of Ae. aegypti within a 24-h period. In the field, a density of 25 adult copepods/400 ml of water in an artificial container prevented the establishment of Aedes mosquitoes by 98% compared to the control container (without copepods). A density of 50 copepods/400 ml of water completely prevented infestation with Aedes. Over a 12-wk period, evaluation of M. brevisetosus-treated ovitraps reduced Ae. aegypti by 38-100% compared to ovitraps without copepods. The introduced gravid copepods increased 2.5-6.7 times over the original cohort. Mesocyclops brevisetosus showed high fecundity in culture, with a mean of 58 nauplii produced per individual gestation cycle.

Animals↗

The aggregation pheromone of Diorhabda elongata, a biological control agent of saltcedar (Tamarix spp.): identification of two behaviorally active components.

The leaf beetle Diorhabda elongata Brullé (Coleoptera: Chrysomelidae) has been introduced as a biological control agent for saltcedars, Tamarix spp., an exotic, invasive weedy tree in the western United State. Gas chromatographic (GC) analysis of volatiles collected from feeding male or female beetles, or saltcedar foliage alone, showed two components produced almost exclusively by males. These compounds elicited responses from antennae of male and female beetles in GC-electroantennographic detection (EAD) analyses. The compounds were identified as (2E,4Z)-2,4-heptadienal (1) and (2E,4Z)-2,4-heptadien-1-ol (2) by GC-mass spectrometry (MS), and confirmed with authentic standards. The two compounds were also detected at trace levels from feeding females and foliage controls, but the amounts from feeding males were 8-40 times higher, typically 55-125 ng per day per male. The amounts of 1 and 2 in collections from females did not differ significantly from amounts collected from control foliage. In field trials, 2 as a single component was as attractive as a 1:1 blend of 1 and 2. Compound 1 as a single component was more attractive than controls, but much less attractive than 2 or the blend. Males and females were attracted in about equal numbers, indicating that this is an aggregation pheromone.

Aldehydes↗

Cercosporella acroptili and Cercosporella centaureicola sp. nov.--potential biological control agents of Russian knapweed and yellow starthistle, respectively.

Russian knapweed (Acroptilon repens [L.] DC.) and yellow starthistle (Centaurea solstitialis L.) are invasive weeds in the western United States, and both weeds are targeted for biological control. Cercosporella acroptili (Bremer) U. Braun was identified as a possible biological control agent for A. repens, and a morphologically similar Cercosporella sp. recently was found damaging to C. solstitialis in the field. Because both fungi are potentially important for biological control of the respective weeds, studies were undertaken to ascertain whether the isolates were identical based on morphology, pathogenicity, growth and spore production, and genetics (molecular characterization of the internal transcribed spacer regions of the ribosomal RNA genes). Differences in these variables between the two isolates were sufficient to indicate that the isolate from C. solstitialis was distinct and justified a new description at the species level: Cercosporella centaureicola sp. nov.

Ascomycota↗

The use of the copepod Mesocyclops longisetus as a biological control agent for Aedes aegypti in Cali, Colombia.

We present data on the efficacy of Mesocyclops longisetus as a biocontrol agent in controlling Aedes aegypti larvae in catch basins in Cali, Colombia. Additionally, we determined some of the features that facilitated the establishment of the copepods in catch basins. Between June 1999 and February 2000, 201 catch basins were treated with an average of 500 adult copepods. The copepods had established in 49.2% of all the basins and they maintained Ae. aegypti larvae at low densities until the end of the 8-month study. The corrected efficacy percent was 90.4%. The copepods established in basins located in a flat area as opposed to those in steep areas, exposed to sunlight and with 0-10% of floating organic matter. When the catch basins were contaminated with synthetic washing agents, like detergents, the copepods did not survive. The copepod M. longisetus could be incorporated as a biological control agent in an integrated Ae. aegypti control program.

Aedes↗

Oxidative burst elicited by Bacillus mycoides isolate Bac J, a biological control agent, occurs independently of hypersensitive cell death in sugar beet.

Response of sugar beet cultivars C40 and USH11 to syringe infiltration of live and dead Bacillus mycoides isolate Bac J, a biological control agent, and virulent and avirulent isolates of Erwinia carotovora pv. betavasculorum was measured by monitoring systemic acquired resistance control of Cercospora beticola, specific activity of chitinase and beta-glucanase, the oxidative burst, and hypersensitive cell death at the infiltration site. Priming sugar beet with B. mycoides Bac J (1 x 10(8) cells/ml) and avirulent isolates of E. carotovora pv. betavasculorum (1 x 10(6) cells/ml) reduced C. beticola symptoms by nearly 70% on distal, untreated leaves. Systemic resistance responses elicited by live B. mycoides Bac J and avirulent E. carotovora pv. betavasculorum isolates, measured by assays for chitinase and beta-glucanase, were statistically equivalent, and biphasic hydrogen peroxide production was observed. Although similar in timing, the second hydrogen peroxide burst was twofold lower for B. mycoides Bac J than for avirulent E. carotovora pv. betavasculorum. Hypersensitive cell death was elicited by avirulent E. carotovora pv. betavasculorum but not B. mycoides Bac J. An oxidative burst was elicited by spray-applied B. mycoides Bac J under both light and green light conditions, indicating that the signal produced by B. mycoides Bac J was not reliant on the stomata for entry into sugar beet. A working model for signal delivery and systemic resistance induction by B. mycoides Bac J in sugar beet is proposed.

Bacillus↗

Occurrence of Puccinia isiacae on Phragmites australis in Saudi Arabia and its possibility as a biological control agent.

During the summer of 2002, symptoms of rust disease were observed for the first time on Phragmites australis in Saudi Arabia. Light brown lesions of regular shape indicating uredinia of Puccinia isiacae appeared on the leaves. The morphology and characteristics of the fungus were described in detail with both light and scanning electron microscopy. The possibility of using this fungus as a biological control agent was also discussed.

Basidiomycota↗

Ecological Similarity and Coexistence of Epiphytic Ice-Nucleating (Ice) Pseudomonas syringae Strains and a Non-Ice-Nucleating (Ice) Biological Control Agent.

De Wit replacement series were used to study competitive interactions between epiphytic IcePseudomonas syringae strains and the biological frost control agents IceP. syringae TLP2del1 and Pseudomonas fluorescens A506. Mixtures containing two strains in different proportions but at a constant total population size were inoculated onto potato leaves. The population sizes of each strain and the total population size were determined when the community had reached equilibrium. A near-isogenic P. syringae strain pair exhibited an interaction similar to that expected for strains competing equally for limiting environmental resources. Replacement series with nonisogenic Ice and IceP. syringae strain pairs suggested that these strains competed for limiting resources according to their relative competitive abilities. There was no evidence of any niche differentiation between the IceP. syringae strains and the IceP. syringae strain. The growth responses of epiphytes following addition of nutrients to the phyllosphere indicated that the epiphytic P. syringae populations were nutrient limited and that, under growth chamber conditions, the populations were more limited by the availability of carbon than by the availability of nitrogen. Determination of in vitro carbon source utilization profiles provided further evidence for the lack of niche differentiation between the Ice and the IceP. syringae strains. Niche overlap indices calculated for the IceP. syringae strains with respect to IceP. syringae TLP2del1 were uniformly high, indicating ecological similarity, and were consistent with the observed low level of coexistence. The biological frost control agent P. fluorescens A506 replaced P. syringae. This was correlated with a high degree of niche overlap between these species.

Journal Article↗

Comparative toxicity of seven insecticides to immature stages of Musca domestica (Diptera: Muscidae) and two of its important biological control agents, Muscidifurax raptor and Spalangia cameroni (Hymenoptera: Pteromalidae).

The toxicity of seven insecticides was evaluated against unparasitized Musca domestica L. pupae and pupae parasitized by Muscidifurax raptor Girault & Sanders or Spalangia cameroni Perkins, two important biological control agents. Only pyrethrins + piperonyl butoxide (Pyrenone) was less toxic to M. raptor compared with house flies. Conversely, all of the insecticides except crotoxyphos were less toxic to S. cameroni compared with house flies. A plateau in the tetrachlorvinphos bioassay line for S. cameroni suggested that this colony had approximately 45% resistant individuals. The selectivity observed between immature stages of house flies and M. raptor or S. cameroni is different from that reported against adult stages of these same species, suggesting that selectivity of an insecticide varies considerably between different life stages.

Animals↗

Viability and stability of biological control agents on cotton and snap bean seeds.

Cotton and snap bean were selected for a multi-year, multi-state regional (south-eastern USA) research project to evaluate the efficacy of both commercial and experimental bacterial and fungal biological control agents for the management of damping-off diseases. The goal for this portion of the project was to determine the viability and stability of biological agents after application to seed. The biological seed treatments used included: (1) Bacillaceae bacteria, (2) non-Bacillaceae bacteria, (3) the fungus Trichoderma and (4) the fungus Beauveria bassiana. Seed assays were conducted to evaluate the following application factors: short-term (< or = 3 months) stability after seed treatment; quality (i.e. isolate purity); compatibility with chemical pesticides and other biocontrol agents; application uniformity between years and plant species. For the bacterial treatments, the Bacillaceae genera (Bacillus and Paenibacillus) maintained the greatest population of bacteria per seed, the best viability over time and the best application uniformity across years and seed type. The non-Bacillaceae genera Burkholderia and Pseudomonas had the least viability and uniformity. Although Beauveria bassiana was only evaluated one year, the seed fungal populations were high and uniform. The seed fungal populations and uniformity for the Trichoderma isolates were more variable, except for the commercial product T-22. However, this product was contaminated with a Streptomyces isolate in both the years that it was evaluated. The study demonstrated that Bacillaceae can be mixed with Trichoderma isolates or with numerous pesticides to provide an integrated pest control/growth enhancement package.

Bacillaceae↗

Genetic variation and founder effects in the parasitoid wasp, Diaeretiella rapae (M'intosh) (Hymenoptera: Braconidae: Aphidiidae), affecting its potential as a biological control agent.

The effects of recent colonization on the aphid parasitoid, Diaeretiella rapae (M'Intosh) (Hymenoptera: Braconidae: Aphidiidae), in Western Australia were investigated. When compared with populations from the Old World, the results of a microsatellite analysis show that the insects have low allelic length and low allele frequency variation, revealing that these individuals experienced a significant founder effect. Marked genetic differentiation between populations was also revealed, which has potentially important implications for host utilization in this species when introduced to a new geographical area(s). Low genetic variation and gene flow in a founder population could limit evolutionary potential in Australia, including the ability of a population to mount a response to newly introduced hosts, such as the Russian wheat aphid, Diuraphis noxia (Mordvilko). Although the actual importance of genetic diversity in the success of biological control agents is unclear, current theory concerning the potential impact of genetic bottlenecks on additive genetic variance is discussed.

Animals↗

Mass rearing methods for Galerucella calmariensis and G. pusilla (Coleoptera: Chrysomelidae), biological control agents of Lythrum salicaria (Lythraceae).

Purple loosestrife, Lythrum salicaria L., an invasive Eurasian perennial, is degrading wetlands across temperate North America. Because conventional control methods have proven ineffective, current emphasis is on the introduction and release of host-specific biological control agents. To increase the availability of control agents for distribution and thus the potential for faster control, mass rearing techniques for 2 leaf beetles, Galerucella calmariensis L. and G. pusilla Duftschmidt, were developed under greenhouse and field conditions. To evaluate the success of various rearing techniques on the number of beetles produced and the effect of these methods on offspring quality, female fecundity, larval development, and adult survival were monitored. Survival of adults maintained at 4 degrees C was approximately 60% and was independent of the type of material used for storage, duration of overwintering, and number of beetles per container. Survival rates increased with increasing numbers of dry stems offered as an overwintering substrate. Females in smaller cages had significantly higher oviposition rates, and with an increase in the number of beetles per cage, female fecundity doubled. Continuous rearing in the greenhouse reduced female fecundity. Allowing beetles to overwinter significantly improved fecundity and rearing efficiency, independent of whether beetles overwintered in a controlled environment or in the field. Field rearings consistently produced 2-5 times more offspring with higher survival than greenhouse rearings.

Animals↗