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Can synthetic pesticides be replaced with biologically-based alternatives?--an industry perspective.

Agricultural chemical companies have invested in the discovery and development of biological pesticides to complement synthetic pesticides for the control of insects, diseases, and weeds on agronomic and horticultural crops. For plant disease control, companies envisage biological fungicides entering markets where they have the best chance of performing and which are most receptive to using biological control methods. Fewer regulatory requirements can mean faster registration for a biological than a synthetic pesticide. However, industry's requirements for competitive performance, effective formulations, and economic production can mean significant investments in time and money for a biological pesticide, although total investment may be less than for a synthetic pesticide. One biocontrol project in which industry has invested is baculoviruses for insect control. Insect baculoviruses, genetically modified to kill insects faster than wild-type viruses, are attractive biocontrol agents because their selectivity to insect pests and safety to beneficial insects and mammals enable them to compete with synthetic insecticides. Industry is looking for similar biocontrol opportunities in disease control. Biocontrol agents for seedling disease, root rot, and postharvest disease control have been registered by the EPA and are trying to compete with synthetic fungicides for market share. To date, effective biocontrol agents have not been identified for the control of serious foliar diseases, such as grape downy mildew, potato late blight, wheat powdery mildew, and apple scab. Farmers must rely on synthetic fungicides and agronomic methods to control these diseases for the foreseeable future.

Animals↗

Effects of glucosinolates and their enzymatic hydrolysis products via myrosinase on the root-knot nematode Meloidogyne incognita (Kofoid et White) Chitw.

The root-knot nematode Meloidogyne incognita (Kofoid et White) Chitw. is responsible for large yield losses in several horticultural crops. Fumigation with chemicals has been efficient in fighting this soil pest, but it clearly shows a negative environmental impact. Thus, it is necessary to find an environmentally friendly alternative to control this nematode and meet the requirements imposed by world regulation to ban some chemical fumigants in the world after 2005. The glucosinolate-myrosinase system, typical of the Brassicaceae family, appears to be an important natural alternative for the control of several soilborne pests and pathogens. The aim of this study was to evaluate, in vitro, the biocidal activity of 11 glucosinolates and their degradation products on second-stage juveniles of the root-knot nematode M. incognita expressed by the nematicidal (LD(50)) and immobilization effects, after 24 and 48 h. None of the intact glucosinolates had any biological effect. After myrosinase addition, their hydrolysis products (essentially isothiocyanates) resulted in highly different biocidal activities. Among the hydrolysis products of the tested glucosinolates, 2-phenylethyl, benzyl, 4-methylthiobutyl, and prop-2-enyl isothiocyanate showed the stronger activity, with an LD(50) at concentrations of 11, 15, 21, and 34 microM, respectively. On the basis of the in vitro test results, new genotypes of Brassicaceae had been selected for high content in the roots of the glucosinolates generating the more active isothiocyanates and their agronomic performances verified in view of a full-field application as catch crop plants. With this aim, the qualitative and quantitative glucosinolate contents in the roots of these potentially nematicidal plants are also reported and discussed.

Animals↗

[Physiological and biochemical bases of xero-tolerance of fungal biocontrol agents and their application].

Hyphomycetes fungal biocontrol agents usually infect host insects by conidial germination and penetration into insect integument under high humidity condition which is generally the restrictive factor for the application of mycoinsecticides in insect control in the field. It is ideal to produce more xero-tolerant inocula (e.g. conidia) of the agents, although the current formulation technology has helped to some degree to reduce the dependence of fungal formulations on the high humidity. This paper reviewed the advance in basic and applied studies on the physiological and biochemical bases of fungal xero-tolerance and its water activity regulation in the past two decades. Possible approaches to make fungal formulations more xero-tolerant were discussed, with an emphasis on the increased accumulation of biologically compatible solutes such as low-molecular polyols and trehalose to the inocula produced for formulation. More efforts are necessitated to understand the mechanisms involved in molecular biology and in regulating the accumulation of the compatible solutes, and to explore for new technology to enhance the xero-tolerance of the fungal agents through biochemical regulation in mass production.

Fungi↗

Inheritance of resistance to Bt canola in a field-derived population of Plutella xylostella.

Crops expressing Bacillus thuringiensis (Bt) insecticidal Cry proteins are grown on millions of hectares. Recommendations to delay resistance are based on a high expression/refugia strategy that aims to kill resistant heterozygotes and enable some susceptible insects to survive. Leaf-dip bioassays on F1 crosses of Malaysian populations of diamondback moth (Plutella xylostella (L)) showed that CrylAc resistance was not fully recessive. The survival of ca 50% of heterozygotes on Bt canola (Brassica napus L) leaves expressing low concentrations of CrylAc agreed with a non-fully-recessive model for resistance. Extrapolations based on log dose-logit mortality regressions for heterozygotes using leaf-dip bioassays showed that a relatively high level of expression, of ca 2000 ng CrylAc mg(-1) total leaf protein, would be required to give 90% mortality to heterozygotes. If high enough levels of expression of Bt toxin to kill heterozygotes cannot be achieved and maintained under field conditions, the effectiveness of the high-dose/refugia strategy would be reduced.

Animals↗

Extracellular proteases of Trichoderma species. A review.

Cellulolytic, xylanolytic, chitinolytic and beta-1,3-glucanolytic enzyme systems of species belonging to the filamentous fungal genus Trichoderma have been investigated in details and are well characterised. The ability of Trichoderma strains to produce extracellular proteases has also been known for a long time, however, the proteolytic enzyme system is relatively unknown in this genus. Fortunately, in the recent years more and more attention is focused on the research in this field. The role of Trichoderma proteases in the biological control of plant pathogenic fungi and nematodes has been demonstrated, and it is also suspected that they may be important for the competitive saprophytic ability of green mould isolates and may represent potential virulence factors of Trichoderma strains as emerging fungal pathogens of clinical importance. The aim of this review is to summarize the information available about the extracellular proteases of Trichoderma. Numerous studies are available about the extracellular proteolytic enzyme profiles of Trichoderma strains and about the effect of abiotic environmental factors on protease activities. A number of protease enzymes have been purified to homogeneity and some protease encoding genes have been cloned and characterized. These results will be reviewed and the role of Trichoderma proteases in biological control as well as their advantages and disadvantages in biotechnology will be discussed.

Biotechnology↗

Simulium spp. control program in Rio Grande do Sul, Brazil.

Insects of the Simuliidae family have been the object of control in Rio Grande do Sul since the 70s. Their constant attacks became a social-economical problem as well as a problem of Public Health, with serious consequences to men and to the economy of the areas in which the insects develop. At first, the control was done with a chemical larvicide Themephos ABATE 500 E, but an imperfect measuring of outflow to determine the quantity of the product made Simulium spp. resistant to it. From 1983 on, following a study of a new method for the outflow measuring, we started to use a biological larvicide Bacillus thuringiensis serovar israelensis based. The biological control uses the new method in 36.4% of the state area, assisting about 3,500,000 inhabitants.

Animals↗

Virulence of Metarhizium anisopliae (Deuteromycotina: Hyphomycetes) on Anastrepha ludens (Diptera: Tephritidae): laboratory and field trials.

Twenty isolates of the fungus Metarhizium anisopliae (Metsch). Sorkin (Ma) were evaluated to determine their virulence against last instar and adult emergence of Mexican fruit fly, Anastrepha ludens (Loew). Larvae were exposed by immersion in a conidial suspension at a concentration of 10(8) UFC/ml under laboratory conditions. Larvae and pupae cumulative mortality rates ranged from 37.9 to 98.75%. Thirteen isolates caused mortality rates > 83.7%, and their LT50 values ranged from 1.8 to 6.2 d. The Ma2, Ma8, and Ma16 isolates were evaluated at seven different concentrations ranging from 10(1) to 10(7) UFC/ml, showing LC50 values from 3.7 to 4.8 x 10(5) UFC/ml. In a field-cage experiment, 200 ml of a conidial suspension of Ma2, at a concentration of 2.5 x 10(6) UFC/ml, was applied on 2,500 cm2 soil surface (2 x 10(5) UFC/cm2). The fungus reduced adult emergence, 22% fewer adults emerging in a sandy loam soil, and 43% fewer in loam soil, compared with the controls. M. anisopliae may offer a preferable alternative to chemicals as a biological control agent against A. ludens.

Animals↗

Insecticidal plant cyclotides and related cystine knot toxins.

Cyclotides are small disulphide-rich peptides found in plants from the violet (Violaceae), coffee (Rubiaceae) and cucurbit (Cucurbitaceae) families. They have the distinguishing structural features of a macrocyclic peptide backbone and a cystine knot made up of six conserved cysteine residues, which makes cyclotides exceptionally stable. Individual plants express a suite of cyclotides in a wide range of tissue types, including leaves, flowers, stems and roots and it is thought that their natural function in plants is as defence agents. This proposal is supported by their high expression levels in plants and their toxic and growth retardant activity in feeding trials against Helicoverpa spp. insect pests. This review describes the structures and activities of cyclotides with specific reference to their insecticidal activity and compares them with structurally similar cystine knot proteins from peas (Pisum sativum) and an amaranthus crop plant (Amaranthus hypocondriancus). More broadly, cystine knot proteins are common in a wide range of organisms from fungi to mammals, and it appears that this interesting structural motif has evolved independently in different organisms as a stable protein framework that has a variety of biological functions.

Amaranthus↗

Mechanisms and means of detection of biocontrol activity of Pseudozyma yeasts against plant-pathogenic fungi.

Fungi belonging to Pseudozyma spp. represent a small group of yeasts that have drawn limited interest in the scientific literature. However, new research with one species of Pseudozyma, Pseudozyma flocculosa, has demonstrated the potential of this yeast as a biocontrol agent of plant-pathogenic fungi. Based on recent work, it appears that P. flocculosa, a natural inhabitant of the phyllosphere, possesses unique means of defending its ecological niche by producing unusual extracellular fatty acids that are detrimental to, among other fungi, powdery mildews, an important group of plant pathogens. Results from these studies have shown that the fatty acids naturally insert themselves into powdery mildew fungi and cause disorganization of cellular membranes and cell disintegration. Further work with insertional mutagenesis yielded mutants of P. flocculosa that represent valuable biological tools to better understand the properties of the yeast. For instance, preliminary work with mutants having lost their antagonistic properties has led to the isolation of a new metabolite with antifungal activity. Discoveries pertaining to the ecology and mode of action of P. flocculosa may lead to the study of unique metabolic or biological processes in other Pseudozyma spp. that could well release the untapped potential of these misunderstood yeasts.

Antibiosis↗

In vitro assessment of Metarhizium anisopliae isolates to control the cattle tick Boophilus microplus.

Metarhizium anisopliae is a filamentous fungus used for tick control. The in vitro effects of 12 M. anisopliae isolates on engorged Boophilus microplus females were analysed. The most pathogenic isolate (E6S1) caused a 100% death rate when 10(7) spores/ml were used to infect ticks. Isolates of M. anisopliae taken from experimentally infected ticks proved to be more pathogenic than fungus maintained on culture media. A comparison between dsRNA mycovirus-free and infected M. anisopliae isolates suggested that, in general, virus free isolates were more infective. The results showed that the biological control of B. microplus by M. anisopliae infection might constitute an additional method to integrated tick control management.

Animals↗

Burkholderia cepacia: medical, taxonomic and ecological issues.

The increasing challenge posed by multiresistant saprophytes in medical microbiology is strikingly demonstrated by the emergence of Burkholderia (formerly Pseudomonas) cepacia as an opportunist pathogen in immunocompromised patients, particularly individuals with chronic granulomatous disease and cystic fibrosis (CF). Best known previously as a phytopathogen and the cause of soft rot of onions, B. cepacia presents three major problems for the CF community: innate multiresistance to antimicrobial agents; person-to-person transmission of epidemic strains through nosocomial or social contacts; and 'cepacia syndrome', a fulminating fatal pneumonia, sometimes associated with septicaemia, that occurs in approximately 20% of colonised patients, including those with previously mild disease. Accumulated evidence to dispel earlier suggestions that the organism is avirulent and merely a marker of existing lung disease includes: case-controlled studies in CF patients; reports of serious infections in non-CF patients; in-vitro and in-vivo evidence that B. cepacia induces production of pro-inflammatory markers, including the major cytokine TNFalpha; and histopathological evidence that exposure of transgenic CF mice to B. cepacia results in pneumonia. By the early 1990s, the use of selective culture media and DNA-based bacterial fingerprinting confirmed suspicions of epidemic person-to-person spread of B. cepacia. This evidence provided scientific justification for draconian and controversial measures for infection control, in particular, segregation of B. cepacia-colonised patients during treatment at CF centres and their exclusion from social gatherings and national conferences. Recently, molecular analyses of type strains and clinical isolates have revealed that isolates identified previously as B. cepacia belong to at least three distinct species and have increased concern regarding the reliability of current laboratory detection and identification systems. Clarification of the taxonomy of B. cepacia-like organisms and the pathogenic potential of environmental isolates remains a high priority, particularly when the organism's antifungal and degradative properties have created interest in its potential use as a biological control agent to improve crop yields and its use for the bioremediation of contaminated soils.

Allium↗

Biological control in greenhouse systems.

The controlled environment of greenhouses, the high value of the crops, and the limited number of registered fungicides offer a unique niche for the biological control of plant diseases. During the past ten years, over 80 biocontrol products have been marketed worldwide. A large percentage of these have been developed for greenhouse crops. Products to control soilborne pathogens such as Sclerotinia, Pythium, Rhizoctonia and Fusarium include Coniothyrium minitans, species of Gliocladium, Trichoderma, Streptomyces, and Bacillus, and nonpathogenic Fusarium. Products containing Trichoderma, Ampelomyces quisqualis, Bacillus, and Ulocladium are being developed to control the primary foliar diseases, Botrytis and powdery mildew. The development of Pseudomonas for the control of Pythium diseases in hydroponics and Pseudozyma flocculosa for the control of powdery mildew by two Canadian research programs is presented. In the future, biological control of diseases in greenhouses could predominate over chemical pesticides, in the same way that biological control of greenhouse insects predominates in the United Kingdom. The limitations in formulation, registration, and commercialization are discussed, along with suggested future research priorities.

Canada↗

Mosquito vector control and biology in Latin America--a second symposium.

The second Spanish language symposium presented by the American Mosquito Control Association (AMCA) was held as part of the 58th Annual Meeting in Corpus Christi, TX in March 1992. The principal objective, as it was for the 1991 symposium, was to increase and stimulate greater participation in the AMCA by vector control specialists and public health workers from Latin America. This publication includes summaries of 25 individual presentations that were given in Spanish. The symposium included the following topics: biology and chemical control of Aedes aegypti and anopheline vectors of malaria; field and laboratory studies of biological control agents for Aedes aegypti; community participation in the prevention of dengue; and other various aspects of the biology of other medically important arthropods (e.g., Simulium ochraceum, Lutzomyia and Culicoides).

Aedes↗

Antifungal characteristics of a fluorescent Pseudomonas strain involved in the biological control of Rhizoctonia solani.

A plant growth-promoting isolate of a fluorescent Pseudomonas spp. EM85 was found strongly antagonistic to Rhizoctonia solani, a causal agent of damping-off of cotton. The isolate produced HCN (HCN+), siderophore (Sid+), fluorescent pigments (Flu+) and antifungal antibiotics (Afa+). Tn5::lacZ mutagenesis of isolate EM85 resulted in the production of a series of mutants with altered production of HCN, siderophore, fluorescent pigments and antifungal antibiotics. Characterisation of these mutants revealed that the fluorescent pigment produced in PDA and the siderophore produced in CAS agar were not the same. Afa- and Flu- mutants had a smaller inhibition zone when grown with Rhizoctonia solani than the EM85 wild type. Sid- and HCN mutants failed to inhibit the pathogen in vitro. In a pot experiment, mutants deficient in HCN and siderophore production could suppress the damping-off disease by 52%. However, mutants deficient in fluorescent pigments and antifungal antibiotics failed to reduce the disease severity. Treatments with mutants that produced enhanced amounts of fluorescent pigments and antibiotics compared with EM85 wild type, exhibited an increase in biocontrol efficiency. Monitoring of the mutants in the rhizosphere using the lacZ marker showed identical proliferation of mutants and wild type. Purified antifungal compounds (fluorescent pigment and antibiotic) also inhibited the fungus appreciably in a TLC bioassay. Thus, the results indicate that fluorescent pigment and antifungal antibiotic of the fluorescent Pseudomonas spp. EM85 might be involved in the biological suppression of Rhizoctonia-induced damping-off of cotton.

Antibiosis↗

[The biocontrol of container-breeding mosquitoes by using Toxorhynchites].

Container-breeding mosquitoes not only bite people but also transmit many human and animal diseases. The problem with chemical control of these mosquitoes is that the breeding sites are always well dispersed and not easily located, and more manenergy and money are required to treat them individually. Since the adverse effects of pesticides on vector control have been recognized, Toxorhynchites mosquitoes are being reconsidered as a means of reducing the populations of other mosquito species. The bionomic interaction of predator and prey, mass rearing, and some examples of biological control of Toxorhynchites are discussed in this article. Integrated controls combining periodic mass release of gravid Toxorhynchites females and ULV pesticide applications can be used to suppress container-breeding mosquitoes.

Animals↗

Isolation and evaluation of antagonistic bacteria towards the cucurbit powdery mildew fungus Podosphaera fusca.

Powdery mildew is one of the most important limiting factors for cucurbits production in Spain, its management being strongly dependent on chemicals. The aim of this work was to evaluate the possibility of exploiting antagonistic bacteria in the biological control of the cucurbit powdery mildew fungus Podosphaera fusca (syn. Sphaerotheca fusca). Among a collection of bacterial strains isolated from distinct cucurbit powdery mildew diseased plants and rhizospheric soils, four isolates were selected, by means of a screening method based on antibiotic production, and identified as Bacillus spp. These isolates proved to be efficacious in the control of cucurbit powdery mildew in in vitro detached leaves and seedling biocontrol assays, where reductions of disease severity of up to 80% were obtained. Furthermore, bacterial populations on melon leaves remained at similar levels (10(5) cfu cm(-2)) over the 16-day period studied and, as observed by scanning electron microscopy analysis, they were able to establish microcolonies associated with an extracellular matrix, which reveals that these isolates efficiently colonize melon phylloplane. These results indicate that the bacterial isolates selected are promising candidates for biological control agents of cucurbit powdery mildew in southern Spain.

Antibiosis↗

Suitability of two root-mining weevils for the biological control of scentless chamomile, Tripleurospermum perforatum, with special regard to potential non-target effects.

The biology and host range of the two root-mining weevils Diplapion confluens Kirby and Coryssomerus capucinus (Beck), two potential agents for the biological control of scentless chamomile Tripleurospermum perforatum (Mérat) Laínz, were studied in the field in southern Germany and eastern Austria, and in a common garden and under laboratory conditions in Delémont, Switzerland from 1993 to 1999. Both weevils were univoltine, and females started to lay eggs in early spring. Diplapion confluens had three and C. capucinus five instars. Larvae of both species were found in the field from mid-April until the end of July; later instars preferentially fed in the vascular cylinder of the shoot base, root crown or root. Although larvae of both species occupy the same temporal and spatial niche within their host plants, they occurred at all investigated field sites together, and showed a similar distribution within sites. No negative or positive interspecific association was detected. Host-specificity tests including no-choice, single-choice, and multiple-choice tests under confined conditions, as well as tests under field conditions with natural and augmented insect densities revealed that both herbivores were specific to plant species in the tribe Anthemideae. However, their development to mature larva or adult on several cultivated plants, as well as on one plant species native to North America, rendered them unsuitable for field release in North America. It was concluded that to investigate non-target effects reliably, host-specificity tests with biological control agents should be carried out under a variety of conditions, particularly with augmented insect densities, as are expected to occur naturally after release.

Animals↗