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Synergistic interaction between Beauveria bassiana- and Bacillus thuringiensis tenebrionis-based biopesticides applied against field populations of Colorado potato beetle larvae.

Commercial biopesticides based on the fungal pathogen Beauveria bassiana strain GHA and the bacterial pathogen Bacillus thuringiensis tenebrionis were applied alone and in combination (tank mixed) against larval populations of the Colorado potato beetle, Leptinotarsa decemlineata, in small plots of potatoes over three field seasons. Interactions between the two products were evaluated in terms of pest-control efficacy. B. bassiana (formulated as Mycotrol) was applied at low and medium label rates of 1.25 and 2.5 x 10(13) conidia/ha, and B. thuringienis (formulated as Novodor) was applied at low and high label rates of 40.3 and 120.8 x 10(6) Leptinotarsa units/ha. Two weekly applications of the bacterial pesticide alone provided 50-85% control of beetle larvae within 14 days after the initial application, while applications of the mycopesticide alone produced no greater than 25% control. Maximum control, in nearly all tests, was produced by the combination of the two products. The combined treatments produced a statistically significant 6-35% greater reduction in larval populations than would have been predicted had the two biopesticides acted independently. This low-level synergistic interaction was observed during all field seasons and resulted from combinations at all rates, including, in one of two tests, the low rates of each product. These results indicate that B. thuringiensis and B. bassiana have strong potential for integrated biologically based management of Colorado potato beetle.

Animals↗

Nematodes of the genus Pristionchus are closely associated with scarab beetles and the Colorado potato beetle in Western Europe.

Evolutionary developmental biology examines how changes in developmental programmes give rise to developmental and, ultimately, morphological novelty. To this end, comparisons of related but distinct organisms have to be performed. The diplogastrid nematode Pristionchus pacificus has been developed as a satellite system for a detailed comparison of various developmental processes to the model organism Caenorhabditis elegans, a rhabditid nematode. In addition to developmental and genetic studies, a genomic platform has been established to analyse the biology of this organism. However, only little is known about where and how Pristionchus pacificus and its relatives live in the wild. Here we show that nematodes of the genus Pristionchus live in close association with scarabaeoid beetles and the Colorado potato beetle. In total, we generated 371 isogenic female lines from 4242 beetles collected at 25 sampling sites all over Europe. Isogenic female lines were subjected to sequence analysis and mating experiments for species determination. The 371 isolates fell into six species. Two hermaphroditic species account for about 60% of the collected nematodes. We found Pristionchus maupasi almost exclusively on cockchafers and Pristionchus entomophagus predominantly on dung beetles. Colorado potato beetles carried the gonochoristic species Pristionchus uniformis, which was only rarely observed on scarabaeoid beetles. We describe the initial evidence for the association of Pristionchus nematodes with beetles and provide a phylogeny based on sequence analysis of the small subunit ribosomal RNA gene.

Animals↗

The cysteine protease activity of Colorado potato beetle (Leptinotarsa decemlineata Say) guts, which is insensitive to potato protease inhibitors, is inhibited by thyroglobulin type-1 domain inhibitors.

High levels of protease inhibitors are induced in potato leaves by wounding. These inhibitors, when ingested by Colorado potato beetle (Leptinotarsa decemlineata Say) larvae, induce expression of specific proteolytic activities in the gut. Induced protease activities cannot be inhibited by potato inhibitors and thus enable the insects to overcome this defence mechanism of potato plants. The induced aminopeptidase and endoproteolytic activities both have the characteristics of cysteine proteases. Twenty-one protein inhibitors of different structural types have been examined for their ability to inhibit these activities in vitro. Members of the cystatin superfamily were found to be poor inhibitors of the induced endoproteolytic activities, except for the third domain of human kininogen, which was a fairly strong inhibitor (75% inhibition). The strongest inhibition (85%) of induced endoproteolytic activity was obtained using structurally different thyroglobulin type-1 domain-like inhibitors--equistatin and MHC class II-associated p41 invariant fragment. Experiments performed using three synthetic substrates for endoproteases gave similar results and indicate the existence of at least different endoproteolytic enzymes resistant to potato inhibitors. The induced aminopeptidase activity can be inhibited only by stefin family of inhibitors in cystatin superfamily. In in vivo experiments, Colorado potato beetle larvae fed on equistatin-coated potato leaves were strongly retarded in their growth and almost 50% died after 4 days. This demonstrated the potential of equistatin to protect crops from insect attack.

Animals↗

Structure of Colorado potato beetle lipophorin: differential scanning calorimetric and small-angle X-ray scattering studies.

The structure of lipophorin, isolated from hemolymph of the Colorado potato beetle, was investigated by differential scanning calorimetry (DSC) and small-angle X-ray scattering. The DSC heating curves of intact lipophorin showed endothermic peaks that were similar to peaks obtained with the hydrocarbon fraction isolated from this lipophorin. The observed peaks correlated with the transition of the hydrocarbons from an ordered into a more disordered state. Changes in structure of the lipophorin particles with increasing temperature were also observed by small-angle X-ray scattering studies. The structural organization of lipophorin was further elucidated by simulation analysis, using a three-layered symmetrical sphere as a model. These studies revealed that lipophorin from the Colorado potato beetle is a sphere with a maximum diameter of 175 A. The sphere is composed of three radially symmetrical layers of different electron densities. The outer layer (37.5-39.5 A in thickness) is composed of phospholipid, apolipophorin I, and part of apolipophorin II. The middle layer (5-10 A) contains diacylglycerol, the rest of apolipophorin II, and probably beta-carotene. The core of the particle (40-45 A) only contains hydrocarbons. This structure differs from another model, previously proposed for cockroach and locust lipophorins [Katagiri, C., Sato, M., & Tanaka N. (1987) J. Biol. Chem. 262, 15857-15861], in the small size of the middle layer. The volume of the middle layer correlated well with the low diacylglycerol content of this lipophorin.

Animals↗

Colorado potato beetle toxins revisited: evidence the beetle does not sequester host plant glycoalkaloids.

The Colorado potato beetle feeds only on glycoalkaloid-laden solanaceous plants, appears to be toxic to predators, and has aposematic coloration, suggesting the beetle may sequester alkaloids from its host plants. This study tested 4th instars and adults, as well as isolated hemolymph and excrement, to determine if the beetles sequester, metabolize, or excrete alkaloids ingested from their host plants. HPLC analysis showed: that neither the larvae nor the adults sequestered either solanine or chaconine from potato foliage; that any alkaloids in the beetles were at concentrations well below 1 ppm; and that alkaloids were found in the excrement of larvae at approximately the same concentrations as in foliage. Analysis of alkaloids in the remains of fed-upon leaflet halves plus excreta during 24 hr feeding by 4th instars, as compared to alkaloids in the uneaten halves of the leaflets, showed that equal amounts of alkaloids were excreted as were ingested. The aposematic coloration probably warns of a previously-identified toxic dipeptide instead of a plant-derived alkaloid, as the Colorado potato beetle appears to excrete, rather than sequester or metabolize, the alkaloids from its host plants.

Animals↗

Comparison of Perillus bioculatus and Podisus maculiventris (Hemiptera:Pentatomidae) as potential control agents of the Colorado potato beetle (Coleoptera:Chrysomelidae).

In laboratory and microplot consumption tests, spined soldier bug, Podisus maculiventris (Say), ate at least as many Colorado potato beetle, Leptinotarsa decemlineata (Say), eggs and larvae as did the twospotted stink bug, Perillus bioculatus (F.). P. maculiventris also remained in field microplots infected with Colorado potato beetle egg masses for slightly longer than did P. bioculatus. The 2 predators were equally effective at consuming egg masses, reducing densities of beetle larvae, and preventing potato defoliation in small field plots. These results contrast with previous trials, in which P. bioculatus had been more effective. In laboratory trials, larger P. maculiventris nymphs selected larger prey larvae, whereas P. bioculatus nymphs did not discriminate by size of prey; thus, the relative effectiveness of the 2 predators may depend on the life stages of the prey that are present at the time of predator release.

Animals↗

Schizophrenia research: a biennium of progress. Proceedings from the Sixth International Congress on Schizophrenia Research. Colorado Springs, CO, April 12-16, 1997.

The Sixth International Congress on Schizophrenia Research (ICOSR) took place in Colorado Springs, Colorado, April 12-16, 1997, where over 1,000 scientists presented and listened to the latest developments in the search for the cause and treatment of schizophrenia. The ICOSR is sponsored by Maryland Psychiatric Research Center, Case Western Reserve University, and the William K. Warren Foundation. The National Institute of Mental Health and several pharmaceutical companies contributed generously to the meeting. The ICOSR is co-organized by Dr. Carol A. Tamminga, Maryland Psychiatric Research Center, University of Maryland at Baltimore, and Dr. S. Charles Schulz, Case Western Reserve University, Cleveland, Ohio. The William K. Warren Research Award is given to a senior investigator, who has made outstanding contributions to our understanding of schizophrenia. The fifth William K. Warren Research Award was presented to Dr. Philip S. Holzman in recognition of his contributions to the identification of eye-tracking abnormalities as a potential phenotypic marker of the illness and also in recognition of his work as a lifelong mentor for schizophrenia researchers. The ICOSR Young Investigator Awards are presented to junior investigators who have demonstrated the potential to make significant contributions to research on schizophrenia. These awards promote scientific development by enabling these young researchers to attend the meeting. There were 30 Young Investigator Award winners. The ICOSR meeting is organized into four sessions: (1) a morning plenary session; (2) a plenary lecture; (3) a poster session; and (4) concurrent afternoon oral sessions. The morning plenary sessions are comprised of a set of 30-minute lectures, which provide an overview of a particular topic area relevant to schizophrenia research. The plenary lecture is an invited lecture on a basic topic related to current research efforts in schizophrenia. The poster sessions provide a forum for the presentation of prepublication reports of basic and clinical science projects. The afternoon sessions are a collection of approximately 10 focused presentations on current research projects related to a specific topic area. The purpose of this report is to provide an account of the proceedings from the plenary and afternoon oral sessions.

Animals↗

The voyage of an invasive species across continents: genetic diversity of North American and European Colorado potato beetle populations.

The paradox of successful invading species is that they are likely to be genetically depauperate compared to their source population. This study on Colorado potato beetles is one of the few studies of the genetic consequences of continent-scale invasion in an insect pest. Understanding gene flow, population structure and the potential for rapid evolution in native and invasive populations offers insights both into the dynamics of small populations that become successful invaders and for their management as pests. We used this approach to investigate the invasion of the Colorado potato beetle (Leptinotarsa decemlineata) from North America to Europe. The beetles invaded Europe at the beginning of the 20th century and expanded almost throughout the continent in about 30 years. From the analysis of mitochondrial DNA (mtDNA) and amplified fragment length polymorphism (AFLP) markers, we found the highest genetic diversity in beetle populations from the central United States. The European populations clearly contained only a fraction of the genetic variability observed in North American populations. European populations show a significant reduction at nuclear markers (AFLPs) and are fixed for one mitochondrial haplotype, suggesting a single successful founder event. Despite the high vagility of the species and the reduction of genetic diversity in Europe, we found a similar, high level of population structure and low gene flow among populations on both continents. Founder events during range expansion, agricultural management with crop rotation, and selection due to insecticide applications are most likely the causes partitioning genetic diversity in this species.

Analysis of Variance↗

Malingering of memory impairment on the Colorado Priming Test.

OBJECTIVES: We hypothesized that simulated malingerers would show a weaker priming effect and a longer response latency than control patients and patients with a brain injury on a priming test. DESIGN: Participants were compared on their performance on the Colorado Priming Test to replicate and extend the original validation study. METHODS: Of 33 participants, 16 were people with a brain injury and 17 were non-head injured control patients who were also instructed to feign cognitive impairment (simulators N = 17). RESULTS: Up to 88 per cent of simulating malingerers and 75 per cent of head-injured participants were correctly classified. CONCLUSIONS: The Colorado Priming Test may be useful in identifying patients feigning memory impairment.

Adult↗

Improved cultivation systems for isolation of the colorado potato beetle spiroplasma.

In North America, the Colorado potato beetle, Leptinotarsa decemlineata, is often infected with the host-specific, gut-inhabiting Colorado potato beetle spiroplasma (CPBS). CPBS is apparently a commensal, but it may be useful in biocontrol if it can be transformed to express an insect-lethal gene. Difficulty in cultivating the organism, however, has hindered the development of a suitable transformation system. In this study, we eliminated the need for coculturing CPBS with insect cells. CPBS was reliably isolated with the BBL Anaerobic GasPak Jar system (low redox, enhanced CO(inf2)), which was easier to use and less expensive than insect cell coculture methods. A further advantage is a reduction in contaminating insect cell components. Use of anaerobiosis should facilitate early-passage screening of isolates for extrachromosomal elements, for use in gene vector constructs. The unique spiral (decreasing amplitude of coils) morphology of CPBS was preserved by anaerobiosis. The use of low-pH (6.0 to 6.5) media allowed aerobic adaptation of CPBS to M1D and SP-4 broth media. These formulations permitted the first cultivation of CPBS on solid media, an accomplishment that will simplify the selection of molecular transformants. Potato beetles collected at four sites in Poland yielded CPBS strains similar to those previously obtained from populations in North America.

Journal Article↗

Colorado tick fever virus in cell culture. II. Physical and chemical properties.

Trent, Dennis W. (University of Oklahoma School of Medicine, Oklahoma City), and L. Vernon Scott. Colorado tick fever virus in cell culture. II. Physical and chemical properties. J. Bacteriol. 91:1282-1288. 1966.-Heat-inactivation kinetics for Colorado tick fever (CTF) virus grown in L cells indicated that more than one rate constant was involved for inactivation at each exposure temperature. An Arrhenius plot of the data indicated the inactivation rate constants to be dependent on the absolute temperature. The energy of activation, for thermal inactivation of the virus, was 17,289 calories per mole, with the Q(10) being 2.6. The optimal pH range for maintenance of CTF viral infectivity was determined to be 7.5 to 7.8. The infectivity of CTF virus was stable to freezing and thawing in diluents which contained: 50% calf serum, 20% glucose, 20% glycerol, 10% bovine serum albumin, 20 mm glutamine, and 2% gelatin. CTF virus replication was insensitive to inhibition by 5-fluoro-2'-deoxyuridine and 5-bromo-2'-deoxyuridine, whereas herpes simplex virus was markedly inhibited, as reported by others. Actinomycin D inhibited CTF virus replication when cells were pretreated for 24 and 12 hr prior to infection, but not when the inhibitor was added at the time of infection. The nucleic acid of CTF virus appears to be of the ribose type.

Arboviruses↗

Identification of ice-nucleating active Pseudomonas fluorescens strains for biological control of overwintering Colorado potato beetles (Coleoptera: Chrysomelidae).

Laboratory studies were conducted to identify ice-nucleating active bacterial strains able to elevate the supercooling point, the temperature at which freezing is initiated in body fluids, of Colorado potato beetles, Leptinotarsa decemlineata (Say), and to persist in their gut. Adult beetles fed ice-nucleating active strains of Pseudomonas fluorescens, P. putida, or P. syringae at 10(6) or 10(3) bacterial cells per beetle had significantly elevated supercooling points, from -4.5 to -5.7 degrees C and from -5.2 to -6.6 degrees C, respectively, immediately after ingestion. In contrast, mean supercooling point of untreated control beetles was -9.2 degrees C. When sampled at 2 and 12 wk after ingestion, only beetles fed P. fluorescens F26-4C and 88-335 still had significantly elevated supercooling points, indicating that these strains of bacteria were retained. Furthermore, beetle supercooling points were comparable to those observed immediately after ingestion, suggesting that beetle gut conditions were favorable not only for colonization but also for expression of ice-nucleating activity by these two strains. The results obtained from exposure to a single, low dose of either bacterial strain also show that a minimum amount of inoculum is sufficient for establishment of the bacterium in the gut. Persistence of these bacteria in Colorado potato beetles long after ingestion was also confirmed using a polymerase chain reaction technique that detected ice-nucleating active bacteria by virtue of their ina genes. Application of these ice-nucleating active bacteria to elevate the supercooling point of this freeze-intolerant insect pest could significantly reduce their winter survival, thereby reducing local populations and, consequently, crop damage.

Animals↗

Portable trench barrier for protecting edges of tomato fields from Colorado potato beetle (Coleoptera: Chrysomelidae).

Experiments were conducted to test a portable trench barrier composed of an extruded, UV-retarded, PVC plastic trough, designed to allow Colorado potato beetles, Leptinotarsa decemlineata (Say), to enter and become trapped and killed inside. Tests demonstrated that the portable plastic trenches were effective as barriers to Colorado potato beetles as they walked into tomato, Lycopersicon esculentum Mill., fields from overwintering sites in the spring. In field tests, plots that were protected by portable trench barriers had significantly fewer beetles per tomato plant, and lower levels of defoliation. Tomato yields in plots that were protected by portable trench barriers were similar to yields in plots that were protected by insecticide sprays, and significantly higher than plots where beetles were not controlled.

Animals↗

Insecticidal activity of avidin combined with genetically engineered and traditional host plant resistance against Colorado potato beetle (Coleoptera: Chrysomelidae) larvae.

Colorado potato beetle, Leptinotarsa decemlineata (Say), is a destructive pest of potato, Solanum tuberosum (L.), in North America. It is renowned for adapting to insecticides. With the arsenal of effective insecticides decreasing, it is important to consider alternative forms of control. Biotin is an essential coenzyme for insect growth and development. Avidin is a protein found in chicken egg that sequesters biotin and has shown insecticidal properties against a range of insect. We assessed the effectiveness of avidin against the Colorado potato beetle neonates in a no-choice detached leaf bioassay at 0, 17, 34, 51, 102, and 204 microg avidin/ml over 12 d. The LC50 was 136 microg avidin/ml (108-188 95% CL). The combined effects of avidin (136 microg avidin/ml) with Bt-Cry3A or leptines were evaluated with neonates and third instars over 12 and 6 d, respectively. Three potato lines were used: susceptible line, a line engineered to express Cry3A from Bacillus thuringiensis, and a line expressing the natural resistance factor leptines. The addition of avidin at the LC50 concentration significantly reduced consumption by neonates, but it did not affect consumption by third instars feeding on the susceptible line and the leptine line. Survival of neonates feeding on the susceptible line with avidin was significantly reduced compared with the susceptible line. Survival of third instars on the Bt-Cry3A with avidin was significantly reduced after 3 d compared with survival on the Bt-Cry3A, suggesting the addition of avidin may increase susceptibility to Bt-Cry3A.

Animals↗

Response of potato aphid (Homoptera: Aphididae) to synthetic potato-derived Colorado potato beetle (Coleoptera: Chrysomelidae) attractant and natural potato odor.

A recently synthesized kairomone blend, based on the volatiles produced by potato (Solanum spp.) plants, has been demonstrated to be attractive to both adult and larval stages of the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae). It was subsequently formulated in a viscous inert carrier for field applications and showed potential for aggregating beetles in treated areas of the field. We investigated effects of this kairomone formulation on the potato aphid, Macrosiphum euphorbiae (Thomas) (Homoptera: Aphididae). The response of both winged and wingless adults to natural potato foliage and synthetic kairomone was tested in a Y-tube olfactometer. Aphid response to untreated potato foliage, foliage treated with the kairomone blend, and foliage treated with blank inert carrier also was tested in petri dishes. In addition, aphid densities on field plots treated with kairomone and blank inert carrier were compared with the control plots. The untreated potato foliage was found to be attractive to wingless, but not winged, potato aphids. In the olfactometer, the foliage treated with synthetic Colorado potato beetle kairomone was not attractive to either winged or wingless aphids. In petri dishes, aphids avoided leaflets treated with both kairomone formulation and its blank carrier. There was no statistical difference between any treatments compared in the field.

Animals↗

Susceptibility of selected rodent species from Colorado to Borrelia burgdorferi.

To determine the susceptibility of some common Colorado (USA) rodent species to Borrelia burgdorferi, pregnant Peromyscus maniculatus, Tamias minimus, and Spermophilus lateralis were trapped in May 1990 and kept in quarantine until their young were old enough to be used in the experiment. Six to eight 8-wk-old individuals of each of the Colorado species and, for comparison, eight laboratory raised P. leucopus were subcutaneously inoculated with > or = 10(5) spirochetes in 0.1 ml in July 1990. Tissue specimens were collected for isolation from these animals through April 1991. Spirochetes were isolated from blood, ear, bladder, kidney, spleen, liver, and eye in Barbour-Stoener-Kelly (BSK) medium from P. maniculatus, P. leucopus and T. minimus. Spirochetes were isolated from at least one tissue from all of these animals and no isolations were obtained from any of the S. lateralis. Thus, three of the four rodent species tested are susceptible to, and could harbor, B. burgdorferi.

Animals↗

Transgenic-Bt potato plant resistance to the colorado potato beetle affect the aphid parasitoid Aphidius nigripes.

Using the biotechnological plant resistance for herbivore control with less reliance on chemicals in integrated pest management (IPM) programs critically depends on predictable interactions with no-target organisms of various trophic levels. Plant resistance to insect pests based on recombinant Bacillus thuringiensis could interfere with natural enemies of non target pests. Performance of the potato aphid parasitoid Aphidius nigripes was studied on the 'Superior-BT line transgenic for the CryllIA toxin of B. thuringiensis, resistance to the Colorado potato beetle; and none transformed 'Superior' line which served as control. Parasitoid survival was significantly lower on the 'Superior-BT' line compared to control. Adult females were largest on 'Superior' and smallest on BT potatoes. This difference was reflected on parasitoid fecundity, which was lowest on 'Superior-BT', and highest on Superior. The results indicate that factor of potato resistance to the Colorado potato beetle affected the fitness of a parasitold of the aphid Macrosiphum euphorbiae, a secondary pest of potato. The effects on the parasitoid were complex but were generally interpretable in terms of host aphid quality variation among potato lines used as food by the aphids during parasitoid development.

Animals↗

[Enzymatic systems of insecticide detoxication in a population of Colorado beetles resistant to permethrin].

The activity of enzymes providing for permetrin detoxication in the imago resistant natural population of the Colorado potato beetle Leptinotarsa decemlineata was studied with the view of elucidating the biochemical mechanisms of resistance to the insecticide. It was demonstrated that the activity of the main enzymes of insect detoxication, i.e., microsomal monooxygenases, nonspecific esterases and glutathione-S-transferases in the permetrin-resistant population of L-decemlineata is enhanced as compared with the permetrin-sensitive population. It was demonstrated that the inhibitors of microsomal monooxygenases, piperonyl butoxide, and of nonspecific esterases, butifos, significantly increase the sensitivity of the resistant population to permetrin. The experimental results suggest that the activity of microsomal monooxygenases and nonspecific esterases is the main factor which determines the resistance of the Colorado potato beetle to permetrin.

Animals↗