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Snail predation by larvae of Sepedon scapularis Adams (Diptera: Sciomyzidae), a potential biocontrol agent of snail intermediate hosts of schistosomiasis in South Africa.

Malacophagous larvae of the fly Sepedon scapularis Adams were shown experimentally to be effective predators of three species of aquatic pulmonate snails tested as prey: Bulinus africanus (Krauss) an important intermediate host of Schistosoma haematobium (Bilharz), Bulinus tropicus (Krauss) and the invasive species Physa acuta Draparnaud. Survival of S. scapularis larvae from instar to instar was negatively affected by the size of prey snails, since larvae tended to be asphyxiated by the mucous secretions of the snails, or by the larval hydrofuge hairs becoming entangled in snail faeces. In experiments to test the choice of S.scapularis larvae for different species and sizes of snails, B.africanus was significantly disfavoured compared with the other two snail species. Small snails (< 3 mm) were more frequently killed by all three larval instars of S.scapularis. First instars killed few, if any, large snails (> 7 mm), whereas second and third instars preyed effectively on all sizes of snails. Third instars killed significantly more snails than younger instars. For larvae offered only one species of snail as prey, the mean total number of snails killed per larva during its entire development was 49 B.africanus, 45 B.tropicus or 34 P.acuta. It is concluded that S.scapularis is potentially useful as a biological control agent for use against indigenous Bulinus and exotic Physa snails.

Analysis of Variance↗

Possible application of a nematophagous fungus as a biological control agent of parasitic nematodes on commercial sheep farms in South Africa.

Biological control of parasitic nematodes of livestock is currently under development and represents another tool that may be integrated into helminth parasite control strategies. This paper presents a brief introduction to commercial sheep farming in South Africa and currently available nematode parasite control methods. These include the FAMACHA clinical assay, strategies of pasture management, dilution of resistant worm species by introduction of susceptible worms, breed resistant sheep and nutritional supplementation. The purpose of this paper is to outline the principles of biological control using nematophagous fungi and how it may be applied on sheep farms in South Africa.

Animal Husbandry↗

Raising activity of Bacillus thuringiensis var. israelensis against Anopheles stephensi larvae by encapsulation in Tetrahymena pyriformis (Hymenostomatida:Tetrahymenidae).

Toxicity of Bacillus thuringiensis var israelensis (B.t.i.) against surface-feeding mosquito larvae of Anopheles stephensi was enhanced by encapsulation in the protozoan Tetrahymena pyriformis. In the laboratory, larvae died about 8 times faster when exposed to protozoan cells filled with B.t.i. than when exposed to the same concentrations of B.t.i. alone. Best larvicidal activities were achieved with ratios of 1:200-1:500 T. pyriformis cells to B.t.i. spores. The concentration of B.t.i. needed to kill 50% of exposed populations was 4-fold lower with T. pyriformis than with B.t.i. alone in 100 ml-test cups. Toxicity enhancement is very likely a consequence of concentrating B.t.i. insecticidal crystal proteins in T. pyriformis cells and floating them to the water surface in the larval feeding zone. Reduction in the exposure time of B.t.i. to unfavorable field conditions, as a result of the decrease in larval mortality time, might improve the persistence of this biological control agent in nature.

Animals↗

Biological control of mosquito larvae by Guppy fish.

Predation potentiality of a biological control agent, the guppy fish, Poecilia reticulata (= Lebistes reticulatus), was studied over the mosquito larvae under laboratory conditions. The third instar larvae of Culex quinquefasciatus mosquito were used as prey. The average consumption rate was 41.0 per day per fish. The female fish was more active (54.9) than male ones (27.0). The feeding activity was found to be higher at higher water temperature. The study suggests that this fish could be used, after careful field trial, as a promising and sustainable biological control agent in controlling filariasis and other mosquito-borne diseases.

Animals↗

The role of nematophagous fungi in the biological control of nematode parasites of livestock.

The control of nematode parasites of livestock is presently based entirely on anthelmintic treatment and grazing management. On their own, these methods are not sustainable because parasites invariably develop resistance to anthelmintics and because of increasing public concern about chemical residues in livestock products and the environment. Although alternative, non-chemotherapeutic control strategies, such as vaccines and genetic selection for resistance are the focus of considerable research activity worldwide, biological control of nematode parasites has been virtually ignored. The little work that has been done is restricted largely to western Europe and involves virtually just one fungal species, namely Arthrobotrys oligospora. More recent studies indicate other known nematophagous fungal species are more efficient predators of infective larvae in sheep faeces than A. oligospora, and others have a greater capacity to survive passage through the gastrointestinal tract of ruminants. These findings plus the relatively sparse numbers of microbial competitors in the fresh faecal environment, compared with the plant rhizosphere where biological control of plant parasites is proving a particularly intractable problem, engenders optimism that the biological control of animal parasitic nematodes may become a practical reality. Such control will never be a substitute for chemotherapy, where the primary purpose is worm removal from the host, but should be incorporated together with other options into integrated pest management systems to provide sustainable nematode control of livestock into the twenty-first century.

Animals↗

Natural products: repellency and toxicity of wild tomato leaf extracts to the two-spotted spider mite, Tetranychus urticae Koch.

The potential of using phytochemicals from leaves of wild tomato for controlling the two-spotted spider mite, Tetranychus urticae Koch, is explored in this study as a promising alternative to the use of synthetic pesticides. Wild tomato accessions of Lycopersicon hirsutum plants that are not consumed by humans were planted under greenhouse conditions for mass production of leaves. Crude extracts from leaves of three accessions of L. hirsutum, six accessions of L. hirsutum f. glabratum, and one accession each of L. pennellii and L. pimpinellifolium were prepared in chloroform, ethanol and hexane. Two spider mite bioassays, one a measure of antibiosis and the other a measure of repellency, were utilized to determine the acaricidal performance of the crude extracts. The bioassay for antibiosis was a 6-h no-choice test. The bioassay for repellency utilized a ring bioassay. Chloroform leaf extracts of L. hirsutum f. glabratum accessions (PI-251304, PI-134417, PI-134418, and PI-126449) exhibited greatest antibiotic activity on two-spotted spider mites; the hexane extracts exhibited greatest repellency. Extracts from PI-251304, PI-126449, PI-134417, and PI-134418 were especially lethal (chloroform) or repellent (hexane). We investigated differences in chemical composition of the crude leaf extracts that may explain the observed differences in mortality and repellency among the different accessions. Major chemical compounds (alpha -curcumene, alpha -zingiberene, trans-caryophyllene, 2-undecanone, and 2-tridecanone) known to have pesticidal efficacy were detected and quantified in the crude leaf extracts using a gas chromatograph (GC) equipped with a mass spectrometer (GC/MS). Lethality of extracts was mainly associated with the presence of high concentrations of 2-tridecanone; repellency of extracts was mainly associated with the presence of trans-caryophyllene. Leaf extracts of L. hirsutum f. glabratum accessions that contain significant quantities of 2-tridecanone and/or trans-caryophyllene could be useful for managing populations of spider mites, which could reduce reliance on synthetic pesticides.

Animals↗

Common, but complex, mode of resistance of Plutella xylostella to Bacillus thuringiensis toxins Cry1Ab and Cry1Ac.

A field collected population of Plutella xylostella (SERD4) was selected in the laboratory with Bacillus thuringiensis endotoxins Cry1Ac (Cry1Ac-SEL) and Cry1Ab (Cry1Ab-SEL). Both subpopulations showed similar phenotypes: high resistance to the Cry1A toxins and little cross-resistance to Cry1Ca or Cry1D. A previous analysis of the Cry1Ac-SEL showed incompletely dominant resistance to Cry1Ac with more than one factor, at least one of which was sex influenced. In the present study reciprocal mass crosses between Cry1Ab-SEL and a laboratory susceptible population (ROTH) provided evidence that Cry1Ab resistance was also inherited as incompletely dominant trait with more than one factor, and at least one of the factors was sex influenced. Analysis of single pair mating indicated that Cry1Ab-SEL was still heterogeneous for Cry1Ab resistance genes, showing genes with different degrees of dominance. Binding studies showed a large reduction of specific binding of Cry1Ab and Cry1Ac to midgut membrane vesicles of the Cry1Ab-SEL subpopulation. Cry1Ab-SEL was found to be more susceptible to trypsin-activated Cry1Ab toxin than protoxin, although no defect in toxin activation was found. Present and previous results indicate a common basis of resistance to both Cry1Ab and Cry1Ac in selected subpopulations and suggest that a similar set of resistance genes are responsible for resistance to Cry1Ab and Cry1Ac and are selected whichever toxin was used. The possibility of an incompletely dominant trait of resistant to these toxins should be taken into account when considering refuge resistance management strategies.

Alleles↗

Mosquito larvicidal activity of Escherichia coli with combinations of genes from Bacillus thuringiensis subsp. israelensis.

The genes cryIVA and cryIVD, encoding 134- and 72-kDa proteins, respectively, and the gene for a regulatory 20-kDa polypeptide of Bacillus thuringiensis subsp. israelensis (serovar H14) were cloned in all seven possible combinations by the Escherichia coli expression vectors pT7 and pUHE. The four combinations containing cryIVA (cryIVA alone, with cryIVD, with the 20-kDa-protein gene, and with both) displayed high levels of mosquito larvicidal activity in pUHE. The toxicity of the combination of cryIVA and cryIVD, with or without the 20-kDa-protein gene, was higher than has ever been achieved with delta-endotoxin genes in recombinant E. coli. Fifty percent lethal concentrations against third-instar Aedes aegypti larvae for these clones decreased (i.e., toxicity increased) continuously to about 3 x 10(5) cells ml-1 after 4 h of induction. Larvicidal activities, obtained after 30 min of induction, were lower for clones in pT7 and decreased for an additional 3.5 h. Induction of either cryIVD or the 20-kDa-protein gene alone resulted in no larvicidal activity in either pT7 or pUHE20. Cloned together, these genes were slightly toxic in pT7 but not in pUHE20. Five minutes of induction of this combination (cryIVD with the 20-kDa-protein gene) in pT7 yielded a maximal mortality of about 40%, which decreased rapidly and disappeared completely after 50 min. CryIVD is thus apparently degraded in E. coli and partially stabilized by the 20-kDa regulatory protein. Larvicidal activity of the combination of cryIVA and cryIVD was sevenfold higher than that of cryIVA alone, probably because of the cross-stabilization of the polypeptides or the synergism between their activities.

Animals↗

Protection from UV-B damage of mosquito larvicidal toxins from Bacillus thuringiensis subsp. israelensis expressed in Anabaena PCC 7120.

A transgenic strain of the nitrogen-fixing filamentous cyanobacterium Anabaena PCC 7120 protected expressed delta-endotoxin proteins of Bacillus thuringiensis subsp. israelensis from damage inflicted by UV-B, a sunlight component that penetrates Earth's ozone layer. This organism, which serves as a food source to mosquito larvae and could multiply in their breeding sites, may solve the environment-imposed limitations of B. thuringiensis subsp. israelensis as a mosquito biological control agent.

Anabaena↗

Growth of Biomphalaria glabrata populations in the presence of the ampullariid snails Pila ovata, Lanistes carinatus and Marisa cornuarietis.

Adults of three ampullariid species were examined for their ability to affect the population growth of Biomphalaria glabrata (M line strain) under laboratory conditions in which food (lettuce) was supplied ad libitum over an eleven week interval. Lanistes carinatus and Pila ovata were obtained from Kenya, and Marisa cornuarietis originated from the Dominican Republic. The presence of either 3 or 9 L. carinatus or 3 M. cornuarietis per 40 l aquarium did not reduce the population size of B. glabrata below levels attained in control aquaria lacking ampullariids. The presence of 9 M. cornuarietis adults significantly reduced B. glabrata populations by week 10 of the experiment. Presence of 3 or 9 P. ovata per aquarium significantly reduced B. glabrata numbers below control levels starting at week four of the experiment. Aquaria with 9 P. ovata were observed to have fewer B. glabrata egg masses than control aquaria, but the number of egg masses observed was not significantly reduced in aquaria with 3 P. ovata or with 3 or 9 L. carinatus or M. cornuarietis. However, the presence of 3 or 9 adults of each ampullariid species resulted in a significant increase in the percentage of such egg masses that disappeared prior to expected hatching date and that were presumed to have been consumed. The results of his laboratory study suggest that further investigation of the role of ampullariids as biological control agents for pulmonate snails in sub-Saharan Africa should focus on P. ovata.

Animals↗

Bacterial endophytes from seeds of Norway spruce (Picea abies L. Karst).

Endophytic bacteria from wooden plants and especially seed-associated endophytes are not well studied. Fresh seeds collected from four Norway spruce trees (Picea abies) from different locations in the Slovene subalpine region were surface-sterilised and dissected into a seed coat, embryo and endosperm. The presence of endophytes was detected by culturing methods and by direct amplification of the eubacterial 16S rDNA gene. Both approaches identified bacteria from genera Pseudomonas and Rahnella in the Norway spruce seeds. Both are known plant-associated bacteria with growth-promoting properties and biological control potential. We suggest that plant seeds could serve as a vector for transmission of beneficial bacteria.

Antibiosis↗

The potential of nematophagous fungi to control the free-living stages of nematode parasites of sheep: studies with Duddingtonia flagrans.

The nematophagous fungus, Duddingtonia flagrans, isolated from a fresh sheep faecal sample obtained from a farm in northern New South Wales, Australia, was subjected to a number of in vivo investigations in both surgically modified and normal sheep to determine its capacity to survive passage through the gastrointestinal tract. Single and sustained dosing experiments established that between 5 x 10(5) and 10(6) chlamydospores/day resulted in a substantial (> 80%) reduction in the number of infective larvae derived from nematode eggs in faeces. This effect can be maintained if dosing continues. These results demonstrate for the first time the potential of nematophagous fungi to be deployed by means of sustained release technology in the biological control of nematode parasites of livestock.

Animals↗

The control of the free-living stages of Strongyloides papillosus by the nematophagous fungus, Arthrobotrys oligospora.

Two laboratory trials were conducted to determine the effect of the addition of spores (conidia) of the nematophagous fungus, Arthrobotrys oligospora, on the development of the ruminant parasite, Strongyloides papillosus, in cultures of bovine faeces. Both studies showed that at a concentration of 2000 conidia/g faeces virtually eliminated infective larvae (> 99% reduction), following 14 days incubation under ideal conditions (25 degrees C and saturated humidity) for free-living development of this parasite species. In one trial, a high level of control was also observed at a 10-fold decrease in conidia concentration (200 spores/g faeces). This work has demonstrated, in principle, that A. oligospora could provide a practical biological control agent against S. papillosus infecting intensively raised young ruminants in the humid tropics/subtropics.

Animals↗

Biological control of the coffee berry borer Hypothenemus hampei (Coleoptera: Curculionidae) by Phymastichus coffea (Hymenoptera: Eulophidae) in Colombia.

The potential of the eulophid parasitoid Phymastichus coffea LaSalle to control coffee berry borer Hypothenemus hampei (Ferrari) populations under field conditions in Colombia was evaluated. Parasitoid adults were released one, five and nine days after artificial infestations of 90-, 150- and 210-day-old coffee berries with H. hampei females. The position of the beetle inside the berry and the parasitism levels were assessed ten days after each P. coffea release. Parasitism of H. hampei by P. coffea was significantly affected by the age of the berries at the time of infestation, and by the position of the beetle inside the berries. Highest levels of parasitism were recorded in 150-day-old berries (75-85%) and in 90-day-old berries (75%) when P. coffea were released one day after the artificial infestation with H. hampei. In 150-day-old berries, highest levels of parasitism were recorded for H. hampei found in the outer layer of the endosperm followed by beetles penetrating the exocarp. Increasing the time of P. coffea releases after the artificial infestations with H. hampei led to decreased levels of parasitism in beetles attacking 90- and 150-day-old coffee berries. Low levels of parasitism were recorded in H. hampei females infesting older coffee berries because most of the beetles had already constructed galleries deep in the endosperm of the berries, i.e. out of reach of the parasitoid. The potential of P. coffea for biological control of coffee berry borer in Colombia is discussed.

Animals↗

Time-concentration-mortality modeling of the synergistic interaction of Beauveria bassiana and imidacloprid against Nilaparvata lugens.

Interactions between the entomopathogenic fungus, Beauveria bassiana SG8702, and imidacloprid on Nilaparvata lugens (Homoptera: Delphacidae) were studied in laboratory bioassays by spraying suspensions of unformulated conidia (assay 1) and aqueous dilutions of emulsifiable conidia formulation alone (assay 2) or together with imidacloprid at the sub-lethal rates of 0.5, 1.0 and 2.0 microg ml(-1) (assays 3-5). Each assay consisted of five conidia concentrations plus an appropriate control and three replicates, each including 30-40 third-instar nymphs, so as to generate time-concentration-mortality data for modeling analysis. A mineral oil-based emulsion used to formulate B bassiana slightly enhanced fungal activity but had no significant impact on the background mortality of N lugens. On the basis of LC50 estimates and associated variances on days 4-12 after spraying, synergistic interactions of both agents or formulations were determined by estimating relative potencies of assay 2 over assay 1 (1.2-9.0), assay 3 over assay 2 (1.3-1.7), assay 4 over assay 2 (7.5-9.6), assay 5 over assay 2 (22.7-101), assay 4 over assay 3 (3.8-5.8), assay 5 over assay 3 (16.1-61.0), and assay 5 over assay 4 (3.0-10.5). The time-concentration-mortality modeling method was not only mathematically but also biologically robust to evaluate the interactions of B bassiana and imidacloprid on N lugens. Compared with their counterparts, enhanced fungal formulations displayed consistently earlier or greater activities against the pest species based on LC50 and LT50 estimates determined from their time-concentration-mortality relationships. The results highlight a potential for pest control by combined formulation or application of B bassiana and imidacloprid.

Animals↗

Evaluation of Duddingtonia flagrans in reducing infective larvae of Haemonchus contortus in feces of sheep.

Consequences of nematode infections due to Haemonchus contortus are a serious constraint for the sheep industry worldwide. Development of anthelmintic resistance and increasing concern about the impact of anthelmintic use dictate the need of alternative control. Such an alternative is using the nematode trapping fungus Duddingtonia flagrans to reduce infective larvae levels on pasture. Two trials were conducted to determine the effect of D. flagrans in reducing infective larvae (predominantly H. contortus) in feces. The first trial determined the dose effect of D. flagrans in reducing infective larvae in feces. Eighteen ewes were dewormed to remove existing infections and randomly assigned to six treatment groups: 5 x 10(4), 1 x 10(5), 2.5 x 10(5), 5 x 10(5), 1 x 10(6) or no (control) spores of D. flagrans per kg of body weight mixed in their feed for 7 days. Fecal samples were collected daily from these and from infected donor ewes. Feces from individual-treated ewes were mixed with equal amounts of donor ewe feces, theoretically approximating oral dose spore concentrations of 2.5 x 10(4), 5 x 10(4), 1.25 x 10(5), 2.5 x 10(5), 5 x 10(5) and no spores, and were cultured. Across dosages and during the 7 days of fungus feeding, percent reduction of infective larvae ranged from 76.6 to 100.0%. The second trial determined the effect of D. flagrans at the dose of 10(5) spores per kg body weight on reducing infective larvae in feces from naturally infected lambs. Twenty lambs were randomly assigned to either treatment or control groups based on fecal egg count. Treatment lambs were fed spores mixed in feed for 7 days. Feces were collected daily and cultured. During the 7 days of fungus feeding, the percent reduction of infective larvae ranged from 82.8 to 99.7%. Results of these trials demonstrated that the nematode trapping fungus D. flagrans was highly effective in reducing infective larvae in sheep feces and should be considered as a biological control agent for integrated nematode control programs.

Administration, Oral↗

The New World screw-worm fly in north Africa.

The course of the North African outbreak of New World screw-worm myiasis (Cochliomyia hominivorax) since its discovery in 1988 is described. Chemical and biological control measures are reviewed, including the current progress of the 'sterile male' (SIT) eradication programme.

Animals↗