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PubMed2001

Insecticidal activity of selected actinomycete strains against the Egyptian cotton leaf worm Spodoptera littoralis (Lepidoptera: Noctuidae).

Actinomycete strains isolated from soil samples of Saudi Arabia and Egypt were used in the present study to investigate the biological activity of their secondary metabolites on the cotton leaf worm Spodoptera littoralis. Secondary metabolites of 41 strains were given to the newly molted last instar larvae through the food plant (Castor leaves). Many actinomycete strains (58% of all) caused larval mortality ranging from 10-60%. Treatments with strains no. 7, 36, 41 resulted in significant prolongation of larval life (10.0 +/- 1.26, 11.5 +/- 0.7 and 10.6 +/- 1.5 days Vs 9.2 +/- 0.4 of controls). Pupation was affected by the actinomycete treatments (60.1% of all). Success of pupation ranged only from 20 to 80%, depending on the efficiency of the tested secondary metabolites. In contrast, the pupal duration had not been significantly affected but 19.5% of actinomycete strains halted the pupation, because different percentages of pupal deformities were counted (100% pupal deformation after treatment with secondary metabolites of actinomycete strains no. 36). In addition, considerable lethal effect of some actinomycetes were observed on pupae (60, 42.9, 40, 50 and 66.6% pupal mortalities caused by secondary metabolites of strains: 2, 9, 16, 18 and 26, respectively). Secondary metabolites of 34.1% of all actinomycete strains blocked the adult emergence. Depending on the available data, Streptomyces and Streptoverticillum were found the most potent actinomycetes affecting the biological and physiological criteria of the present insect species.

Actinomycetales↗

PubMed2006

Induction of plant defense enzymes and phenolics by treatment with plant growth-promoting rhizobacteria Serratia marcescens NBRI1213.

In greenhouse experiments, plant growth-promoting rhizobacteria (PGPR) Serratia marcescens NBRI1213 was evaluated for plant growth promotion and biologic control of foot and root rot of betelvine caused by Phytophthora nicotianae. Bacterization of betelvine (Piper betle L.) cuttings with S. marcescens NBRI1213 induced phenylalanine ammonia-lyase, peroxidase, and polyphenoloxidase activities in leaf and root. Qualitative and quantitative estimation of phenolic compounds was done through high-performance liquid chromatography (HPLC) in leaf and root of betelvine after treatment with S. marcescens NBRI1213 and infection by P. nicotianae. Major phenolics detected were gallic, protocatechuic, chlorogenic, caffeic, ferulic, and ellagic acids by comparison of their retention time with standards through HPLC. In all of the treated plants, synthesis of phenolic compounds was enhanced compared with control. Maximum accumulation of phenolics was increased in S. marcescens NBRI1213-treated plants infected with P. nicotianae. In a greenhouse test, bacterization using S. marcescens NBRI1213 decreased the number of diseased plants compared with nonbacterized controls. There were significant growth increases in shoot length, shoot dry weight, root length, and root dry weight, averaging 81%, 68%, 152%, and 290%, respectively, greater than untreated controls. This is the first report of PGPR-mediated induction of phenolics for biologic control and their probable role in protecting betelvine against P. nicotianae, an important soil-borne phytopathogenic fungus.

Hydroxybenzoates↗

PubMed2003

Top dressing of feed with desiccated chlamydospores of Duddingtonia flagrans for biological control of the pre-parasitic stages of ovine Haemonchus contortus.

Feeding trials were conducted with stall-fed sheep parasitized with Haemonchus contortus. For 10 days they were offered 250 g of a concentrate feed that had been top-dressed with desiccated chlamydospores of Duddingtonia flagrans at 1 x 10(5), 5 x 10(5), 1 x 10(6) or 2 x 10(6) chlamydospores/kg body weight. Pooled faeces from each group on day 7 of spore feeding were spread on different pasture plots. On day 28 after the start of spore feeding, further pooled faeces from each group were spread on the same plots. The larval burdens on the plots were monitored for 2 months and the larval harvest from in vitro faecal cultures were monitored regularly. The application of 1 x 10(6) or more spores/kg body weight virtually eliminated larvae from both the pasture and the faecal cultures. The application of as few as 1 x 10(5) spores/kg body weight had a profound impact on larval recovery. The effect persisted while the spores were being fed but not for more than 4 days following discontinuation of spore feeding. Top dressing supplementary feed with dried chlamydospores offers a potential way of using D. flagrans for biological control of the pre-parasitic stages of H. contortus.

Acetyltransferases↗

PubMed2000

Cytological effects of cellulases in the parasitism of Phytophthora parasitica by Pythium oligandrum.

The ubiquitous oomycete Pythium oligandrum is a potential biocontrol agent for use against a wide range of pathogenic fungi and an inducer of plant disease resistance. The ability of P. oligandrum to compete with root pathogens for saprophytic colonization of substrates may be critical for pathogen increase in soil, but other mechanisms, including antibiosis and enzyme production, also may play a role in the antagonistic process. We used transmission electron microscopy and gold cytochemistry to analyze the intercellular interaction between P. oligandrum and Phytophthora parasitica. Growth of P. oligandrum towards Phytophthora cells correlated with changes in the host, including retraction of the plasma membrane and cytoplasmic disorganization. These changes were associated with the deposition onto the inner host cell surface of a cellulose-enriched material. P. oligandrum hyphae could penetrate the thickened host cell wall and the cellulose-enriched material, suggesting that large amounts of cellulolytic enzymes were produced. Labeling of cellulose with gold-complexed exoglucanase showed that the integrity of the cellulose was greatly affected both along the channel of fungal penetration and also at a distance from it. We measured cellulolytic activity of P. oligandrum in substrate-free liquid medium. The enzymes present were almost as effective as those from Trichoderma viride in degrading both carboxymethyl cellulose and Phytophthora wall-bound cellulose. P. oligandrum and its cellulolytic enzymes may be useful for biological control of oomycete pathogens, including Phytophthora and Pythium spp., which are frequently encountered in field and greenhouse production.

Carboxymethylcellulose Sodium↗

PubMed2000

Identification of a universally primed-PCR-derived sequence-characterized amplified region marker for an antagonistic strain of Clonostachys rosea and development of a strain-specific PCR detection assay.

We developed a PCR detection method that selectively recognizes a single biological control agent and demonstrated that universally primed PCR (UP-PCR) can identify strain-specific markers. Antagonistic strains of Clonostachys rosea (syn. Gliocladium roseum) were screened by UP-PCR, and a strain-specific marker was identified for strain GR5. No significant sequence homology was found between this marker and any other sequences in the databases. Southern blot analysis of the PCR product revealed that the marker represented a single-copy sequence specific for strain GR5. The marker was converted into a sequence-characterized amplified region (SCAR), and a specific PCR primer pair was designed. Eighty-two strains, isolated primarily from Danish soils, and 31 soil samples, originating from different localities, were tested, and this specificity was confirmed. Two strains responded to the SCAR primers under suboptimal PCR conditions, and the amplified sequences from these strains were similar, but not identical, to the GR5 marker. Soil assays in which total DNA was extracted from GR5-infested and noninoculated field soils showed that the SCAR primers could detect GR5 in a pool of mixed DNA and that no other soil microorganisms present contained sequences amplified by the primers. The assay developed will be useful for monitoring biological control agents released into natural field soil.

DNA Primers↗

PubMed1998

Transmission of Leptospira interrogans serovar Balcanica infection among socially housed brushtail possums in New Zealand.

Leptospira interrogans serovar balcanica is a potential vector being investigated for spreading a biological control agent among introduced brushtail possums (Trichosurus vulpecula) in New Zealand. As previous studies have shown that possums are unlikely to contract leptospirosis through a contaminated environment alone, the objective was to determine whether L. interrogans serovar balcanica could be transmitted between sexually mature, socially housed possums. Possums were infected experimentally with L. interrogans serovar balcanica and housed in pairs or groups with uninfected possums for either 70 or 140 days, during the breeding or non-breeding seasons. No transmission occurred between any infected and uninfected possums during the non-breeding season. However, transmission occurred between females that had been socially housed in pairs or groups in the breeding season. Mixed sex transmission also occurred in pairs and groups, both from males to females and from females to males. Mixed sex transmission usually occurred rapidly (< 44 days) and was not associated with the production of offspring. No transmission occurred between males during the breeding or the non-breeding seasons. Transmission probably occurs as a result of affiliative or sexual behaviour, but is unlikely to occur through fighting. The social transmission pathways determined in this study suggest that L. interrogans serovar balcanica may have the transmission attributes desired in a vector for biological control.

Animals↗

PubMedDate not supplied

[Biological control of helminths in grazing animals using nematophagous fungi].

For the last 50 years the control of gastro-intestinal nematodes (GIN) in grazing animals has almost entirely been alleviated by the use of anthelmintics. Due to development of resistance against the drugs, especially in the GIN of sheep and goats it has become necessary to develop new, innovative strategies such as the use of nematode destroying fungi. Despite experiments to employ various species against plant and animal parasitic nematodes were already attempted in the 1930's, it was not until the 1990's when selection by simulating passage through the gastro-intestinal tract of cattle led to isolation of the fungus Duddingtonia flagrans that a major breakthrough was achieved. This fungus, producing sticky three-dimensional network and now isolated world-wide, is special due to its capacity to prolifically produce high numbers of thick-walled resting spores, chlamydospores. These spores survive passage through the gastro-intestinal tract of grazing livestock and are capable of growing and subsequently trap nematodes, including larval stages of parasitic nematodes. The great potential of this fungus as a biological control agent has been demonstrated through numerous trials with cattle, sheep, horses, and pigs. But these trials have also pointed towards some potential limitations in the activity spectrum of the fungus (Dictyocaulus, Nematodirus) beside the whole group of parasites spreading through infective stages protected inside resistant eggs (e.g. Ascaris, Trichuris). So far, in the few reported studies conducted, no negative environmental impact has been found, but it is important that further studies are conducted on this important issue. Although the potential use of D. flagrans chlamydospores has been verified through numerous trials it is necessary to develop practical delivery systems such as slow release devices, feed-blocks or similar to be able to implement this tool in future integrated control strategies. Such control strategies could include the use of biological control, grazing management, smart use of existing drugs, parasite resistant animal breeds, bioactive forages, and possibly vaccines.

Animals↗

PubMed2005

Functional GNA expressed in Escherichia coli with high efficiency and its effect on Ceratovacuna lanigera Zehntner.

The mannose-specific GNA (Galanthus nivalis agglutinin, snowdrop lectin) are the resistant proteins with many bioactivities. Snowdrop lectin is different with plant organs and development periods in lectin species, content, and bioactivities. It is an effective and cheap way to obtain much active GNA through overexpression of GNA gene in Escherichia coli. Constructs encoding mature GNA fused with an N-terminal pelB signal sequence protein (PelB) were expressed in E. coli with high efficiency. Recombinant protein productivity was higher than values published before. The insecticidal activity of purified recombinant proteins was assayed on feeding sugarcane wooly aphid (Ceratovacuna lanigera Zehntner), as well as spraying on sugarcane plants infected by aphids. The insecticidal activity was found to be comparable to native GNA. Oral delivery has obvious positive implications for crop protection against insect pests since peptides can be present in, or sprayed on, plant tissues susceptible to damage. A highly efficient expression of functional recombinant GNA would decrease the cost of GNA and promote its wide use, especially to give crop protection in the field.

Amino Acid Sequence↗

PubMed2006

Field resistance of codling moth against Cydia pomonella granulovirus (CpGV) is autosomal and incompletely dominant inherited.

The current appearance of local codling moth populations with resistance to Cydia pomonella granulovirus (CpGV) is an impediment to continuous CpGV application. Therefore, crossing experiments have been performed in order to gain information about the inheritance of resistance. Evidence is presented that the observed field resistance is stably inherited even under non-selective conditions in the laboratory. Offspring of reciprocal F(1) crosses between a susceptible ('S') and a resistant ('R') strain and backcrosses between F(1) and S were bioassayed at different CpGV concentrations. The resistant strain showed 100 times lower susceptibility in 7-day bioassays. The responses of the reciprocal crosses (male S x female R and female S x male R) did not differ significantly, indicating that resistance is autosomally inherited. The median lethal concentration for the F(1) progeny was intermediate between those of its parental strains. Mortality data obtained from the backcrosses suggested that inheritance of resistance is due to a non-additive, polygenic trait.

Animals↗

PubMed1999

Efficient expression of mosquito-larvicidal proteins in a gram-negative bacterium capable of recolonization in the guts of Anopheles dirus larva.

The gram-negative bacterium, An11/2 G1, isolated from the guts of Anopheles dirus mosquito larvae, was identified as Enterobacter amnigenus. The E. amnigenus was able to recolonize in the gut of An. dirus larva but not in those of Aedes aegypti and Culex quinquefasciatus larvae. It was able to float in water for a longer period than Bacillus thuringiensis subsp. israelensis and Bacillus sphaericus. These are desirable characteristics for a delivery vehicle of mosquito-larvicidal toxins for the control of mosquito larvae, and E. amnigenus was therefore used as a host to express the cryIVB gene of B. thuringiensis subsp. israelensis and the binary toxin genes of B. sphaericus. The recombinant E. amnigenus produced a high level of CryIVB protein, which was toxic to larvae of Ae. aegypti and An. dirus. Another E. amnigenus producing the 51-kDa protein of B. sphaericus was toxic to larvae of An. dirus and Cx. quinquefasciatus. The recombinant plasmids were stable in E. amnigenus without the presence of selective pressure for at least 23 generations. The recombinant E. amnigenus should represent a desirable biological agent for controlling mosquito larvae.

Animals↗

PubMed2002

Secretion of beta-1,3-glucanases by the yeast Pichia membranifaciens and its possible role in the biocontrol of Botrytis cinerea causing grey mold disease of the grapevine.

Pichia membranifaciens strain FY-101, isolated from grape skin, was found to be antagonistic to Botrytis cinerea, the causal organism of the grey mold disease of the grapevine. When grown together on solid as well as in liquid media, the yeast brings about the inhibition of Botrytis cinerea, which in turn loses its ability to produce the grey mold symptoms on the grapevine plantlets. The secretion of beta-1,3-glucanases by P. membranifaciens is one of the possible mechanisms related to this antagonism. In vitro experiments confirm that this yeast can be used as a biological control organism against B. cinerea. An account of this antagonism and the production of beta-1,3-glucanases by P. membranifaciens is given here.

Botrytis↗

PubMed1995

Biological control of trichostrongyles in calves by the fungus Duddingtonia flagrans fed to animals under natural grazing conditions.

The present study was conducted in the 1993 grazing season with yearling calves exposed to a pasture with a natural mixed trichostrongyle larval infection. It was shown that daily feeding with the microfungus Duddingtonia flagrans during the first 2 months of the season led to a lowered herbage infectivity and a reduced acquisition of Ostertagia sp. and Cooperia sp. later in the season. In addition, the procedure delayed the onset of clinical disease. This was due to the nematode-destroying effects of the fungi in the dung excreted by the fungus-treated calves, as evidenced by results from a parallel in vitro assay on faecal larval cultures. The paper discusses future research needs before practical biological control can be implemented.

Animal Feed↗

PubMed2006

Insecticidal toxins from Photorhabdus bacteria and their potential use in agriculture.

Most of the insecticidal toxins used in agriculture come from a single bacterium Bacillus thuringiensis or 'Bt'. Here we review our work on the array of toxins produced by Photorhabdus and Xenorhabdus bacteria that are symbiotic with entomopathogenic nematodes, and discuss their potential for use in agriculture as alternatives to Bt. Despite the fact that both Photorhabdus and Xenorhabdus are introduced directly into the insect blood stream by their nematode vectors, they produce a range of toxins with both oral and injectable insecticidal activity. The toxin complexes (Tc's) are large orally active toxins that are displayed on the outer surface of the bacterium. They require three components (A-C) for full toxicity and one 'A' component has been successfully expressed in transgenic Arabidopsis to confer insect resistance. One such group of Tc's, the PirAB binary toxins, have oral activity against mosquitoes and some caterpillar pests. Their mode of action is not known but they show significant sequence similarity to a recently described neurotoxin beta-leptinotarsin-h isolated from the blood of the Colorado potato beetle. Other toxins such as 'makes caterpillars floppy' (Mcf) and proteins encoded by the 'Photorhabdus virulence cassettes' (PVCs) only show injectable activity. Mcf1 promotes apoptosis in a wide range of cells and appears to mimic mammalian BH3 domain-only proteins in the mitochondrion whereas the mode of action of the PVCs remains undetermined. The likely biological reasons for the massive functional redundancy in Photorhabdus insecticidal toxins are discussed.

Agriculture↗

PubMed2006

Persistence and efficacy of Beauveria brongniartii strains applied as biocontrol agents against Melolontha melolontha in the Valley of Aosta (northwest Italy).

AIMS: To monitor and select genetically characterized strains of Beauveria brongniartii to be used as microbiological control agents against Melolontha melolontha in different climatic conditions of the Valley of Aosta (northwest Italy). METHODS AND RESULTS: Molecular random amplified polymorphic DNA markers allowed monitoring of five B. brongniartii strains (C2, F, K2, N3 and W2) in field trials. Ten sites were chosen at Jovençan, Saint-Pierre and Quart areas, where a mixture of the five strains colonizing rye kernels was applied to the soil of each M. melolontha infested site. Growth, persistence and virulence on M. melolontha larvae of five fungal strains were evaluated in two subsequent 24-month studies. Beauveria brongniartii grew best at the Jovençan sites. Not only did strain F persist better than the other strains in most soil samples but it was also the most virulent strain. Strain F was isolated the most frequently from infected M. melolontha larvae recovered from the test sites. A general decrease in the larvae rate was detected in the test field soil. CONCLUSIONS: Strain F of B. brongniartii was better than other strains in growth, persistence and virulence against M. melolontha larvae in the test site soil. SIGNIFICANCE AND IMPACT OF THE STUDY: Results obtained from preliminary field trials support the use of strain F as a biological control agent against M. melolontha in the Valley of Aosta even if further targeted studies are still necessary.

Animals↗

PubMed2006

Assessment of the environmental fate of the biological control agent of fire blight, Pseudomonas fluorescens EPS62e, on apple by culture and real-time PCR methods.

The colonization of apple blossoms and leaves by Pseudomonas fluorescens EPS62e was monitored in greenhouse and field trials using cultivable cell counting and real-time PCR. The real-time PCR provided a specific quantitative method for the detection of strain EPS62e. The detection level was around 10(2) cells g (fresh weight)(-1) and the standard curve was linear within a 5-log range. EPS62e actively colonized flowers reaching values from 10(7) to 10(8) cells per blossom. In apple flowers, no significant differences were observed between population levels obtained by real-time PCR and plating, suggesting that viable but nonculturable (VBNC) cells and residual nondegraded DNA were not present. In contrast, on apple leaves, where cultivable populations of EPS62e decreased with time, significant differences were observed between real-time PCR and plating. These differences indicate the presence of VBNC cells or nondegraded DNA after cell death. Therefore, the EPS62e population was under optimal conditions during the colonization of flowers but it was stressed and poorly survived on leaves. It was concluded that for monitoring this biological control agent, the combined use of cultivable cell count and real-time PCR is necessary.

Colony Count, Microbial↗

PubMed2006

Thiamine-auxotrophic mutants of Pseudomonas fluorescens CHA0 are defective in cell-cell signaling and biocontrol factor expression.

In the biocontrol strain Pseudomonas fluorescens CHA0, the Gac/Rsm signal transduction pathway positively controls the synthesis of antifungal secondary metabolites and exoenzymes. In this way, the GacS/GacA two-component system determines the expression of three small regulatory RNAs (RsmX, RsmY, and RsmZ) in a process activated by the strain's own signal molecules, which are not related to N-acyl-homoserine lactones. Transposon Tn5 was used to isolate P. fluorescens CHA0 insertion mutants that expressed an rsmZ-gfp fusion at reduced levels. Five of these mutants were gacS negative, and in them the gacS mutation could be complemented for exoproduct and signal synthesis by the gacS wild-type allele. Furthermore, two thiamine-auxotrophic (thiC) mutants that exhibited decreased signal synthesis in the presence of 5 x 10(-8) M thiamine were found. Under these conditions, a thiC mutant grew normally but showed reduced expression of the three small RNAs, the exoprotease AprA, and the antibiotic 2,4-diacetylphloroglucinol. In a gnotobiotic system, a thiC mutant was impaired for biological control of Pythium ultimum on cress. Addition of excess exogenous thiamine restored all deficiencies of the mutant. Thus, thiamine appears to be an important factor in the expression of biological control by P. fluorescens.

Amino Acid Sequence↗

PubMed2005

Biological control potential of the obligate parasite Pasteuria penetransagainst the root-knot nematode, Meloidogyne incognita infestation in Brinjal.

The efficacy of the obligate bacterial parasite, Pasteuria penetrans against the rootknot nematode, Meloidogyne incognita infestation was assessed in brinjal. The seedling pans with sterilized soil were inoculated with nematodes and root powder of P. penetrans were applied at different dosages viz., 0 x 10(6), 0.5 x 10(6) spores and 1 x 10(6) spores/pan. Seeds of brinjal cv Co2 were sown in the pans and seedlings were allowed to grow. The seedlings were transplanted to microplots containing sterilized soil. Observations on nematode infestation and plant growth were recorded at seedling, flowering, and fruiting stages. Nematode infestation was significantly reduced by P. penetrans treatment. There was 22, 75 and 86% reduction in nematode population of soil over control at seedling, flowering and fruiting stages, respectively, at higher spore density (1 x 10(6)). Egg mass production was decreased by 63, 78 and 89% over control at 35 (seedling), 100 (flowering) and 160 (fruiting) days after sowing respectively, at 1 x 10(6) spores treated soil. The parasitizing ability of P. penetrans increased with the age of the crop. At higher spore density the percentage of parasitization was increased from 52.0 (35 days after sowing) to 90.0 (160 days after sowing) %. At these stages of the crop, the spore load per juvenile also increased at the higher dose. The P. penetrans application enhanced the plant growth. The weight of the shoot was increased by 17.6% whereas root weight by 41.0% over the control at fruiting stage. The experimental results revealed the potential use of P. penetrans as biological control agent of M. incognita. Application of P. penetrans spores in the nursery is a good strategy since the mass multiplication is quite difficult.

Animals↗

PubMed1985

The food of the larvivorous fish Gambusia affinis (Baird and Girard) and Oreochromis (formerly Tilapia) niloticus (Linnaeus) in Gezira irrigation canals.

The food of both Gambusia affinis and Oreochromis niloticus was studied. Organisms eaten by both species of fish are tabulated, together with the amounts eaten during the various months of the year. G. affinis larger than 25 mm are carnivorous and become more so with age. Food selection by G. affinis depends on the availability of food items rather than choice. It showed a marked preference for mosquito larvae. O. niloticus smaller than 150 mm were markedly carnivorous, but this decreased with age. Only small fish of this species are useful for the biological control of mosquitoes. Fish larger than 150 mm showed a marked preference for higher aquatic macrophytes.

Agriculture↗