Properties of a unique mutant of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus that exhibits a partial many polyhedra and few polyhedra phenotype on extended serial passaging in suspension cell cultures

Serial passaging of wild-type Helicoverpa armigera, single-nucleocapsid (HaSNPV) in H. zea (HzAMI) illsect Cell Cultures results ill rapid selection for the few polyhedra (FP) phenotype. A unique HaSNPV mutant (ppC19) was isolated through plaque purification that exhibited a partial many polyhedra (MP) and FP phenotype. Oil serial passaging in suspension cell cultures, ppC19 produced fivefold more polyhedra than a typical FP mutant (FP8AS) but threefold less polyhedra than the wild-type virus. Most importantly, the polyhedra of ppC19 exhibited MP-like virion occlusion. Furthermore, ppC19 produced the same amount of budded virus (BV) as the FP mutant, which was fivefold higher than that of the wild-type virus. This selective advantage was likely to explain its relative stability in polyhedra production for six passages when compared with the wild-type Virus. However, subsequent passaging of ppC19 resulted in a steel) decline in both BV and polyhedra yields, which was also experienced by FP8AS and the wild-type virus Lit high passage numbers. Genomic deoxyribonueleic Licid profiling of the latter suggested that defective interfering particles (DIPS) were implicated in this phenomenon and represented another Undesirable mutation during serial passaging of HaSNPV Hence, a strategy to isolate HaSNPV Clones that exhibited MP-like polyhedra production but FP-like BV production, coupled with low multiplicities of infection during scale-up to avoid accumulation of DIPS, could prove commerically invaluable.

Helicoverpa armigera (Hubner): can wheat stubble protect cotton plants against attack?

When investigating strategies for Helicoverpa armigera (Hubner) control, it is important to understand oviposition behaviour. Cotton (Gossypium hirsutum) was sown into standing wheat (Triticum astivum L.) stubble in a closed arena to investigate the effect of stubble on H. armigera moth behaviour and oviposition. Infrared cameras were used to track moths and determine whether stubble acted as a physical barrier or provided camouflage to cotton plants, thereby reducing oviposition. Searching activity was observed to peak shortly before dawn (03:00 and 04:00 h) and remained high until just after dawn (4 h window). Moths spent more time resting on cotton plants than spiralling above them, and the least time flying across the arena. While female moths spent more time searching for cotton plants growing in wheat stubble, the difference in oviposition was not significant. As similar numbers of eggs were laid on cotton plants with stubble (3.5/plant SE +/- 0.87) and without stubble (2.5/plant SE +/- 0.91), wheat stubble does not appear to provide camouflage to cotton plants. There was no significant difference in the location of eggs deposited on cotton plants with and without stubble, although more eggs were laid on the tops of cotton leaves in wheat stubble. As the spatial and temporal distribution of eggs laid on the cotton plant is a crucial component of population stability, eggs laid on the upper side of leaves on cotton plants may be more prone to fatalities caused by environmental factors such as wind and rain. Therefore, although stubble did not influence the number of eggs laid, it did affect their distribution on the plant, which may result in increased mortality of eggs on cotton plants sown into standing wheat stubble.

Predicting population dynamics of weed biological control agents: science or gazing into crystal balls?

Various factors can influence the population dynamics of phytophages post introduction, of which climate is fundamental. Here we present an approach, using a mechanistic modelling package (CLIMEX), that at least enables one to make predictions of likely dynamics based on climate alone. As biological control programs will have minimal funding for basic work (particularly on population dynamics), we show how predictions can be made using a species geographical distribution, relative abundance across its range, seasonal phenology and laboratory rearing data. Many of these data sets are more likely to be available than long-term population data, and some can be incorporated into the exploratory phase of a biocontrol program. Although models are likely to be more robust the more information is available, useful models can be developed using information on species distribution alone. The fitted model estimates a species average response to climate, and can be used to predict likely geographical distribution if introduced, where the agent is likely to be more abundant (i.e. good locations) and more importantly for interpretation of release success, the likely variation in abundance over time due to intra- and inter-year climate variability. The latter will be useful in predicting both the seasonal and long-term impacts of the potential biocontrol agent on the target weed. We believe this tool may not only aid in the agent selection process, but also in the design of release strategies, and for interpretation of post-introduction dynamics and impacts. More importantly we are making testable predictions. If biological control is to become more of a science making and testing such hypothesis will be a key component.

Temporal variation in arthropod sampling effectiveness: the case for using the beat sheet method in cotton

Predatory insects and spiders are key elements of integrated pest management (IPM) programmes in agricultural crops such as cotton. Management decisions in IPM programmes should to be based on a reliable and efficient method for counting both predators and pests. Knowledge of the temporal constraints that influence sampling is required because arthropod abundance estimates are likely to vary over a growing season and within a day. Few studies have adequately quantified this effect using the beat sheet, a potentially important sampling method. We compared the commonly used methods of suction and visual sampling to the beat sheet, with reference to an absolute cage clamp method for determining the abundance of various arthropod taxa over 5 weeks. There were significantly more entomophagous arthropods recorded using the beat sheet and cage clamp methods than by using suction or visual sampling, and these differences were more pronounced as the plants grew. In a second trial, relative estimates of entomophagous and phytophagous arthropod abundance were made using beat sheet samples collected over a day. Beat sheet estimates of the abundance of only eight of the 43 taxa examined were found to vary significantly over a day. Beat sheet sampling is recommended in further studies of arthropod abundance in cotton, but researchers and pest management advisors should bear in mind the time of season and time of day effects.

Oviposition and reproductive performance of a generalist parasitoid (Trichogramma pretiosum) exposed to host species that differ in their physical characteristics

The response of generalist egg parasitoids to alternative natural hosts that are present simultaneously is not well known. We investigated the behavior of Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) in relation to two field hosts Helicoverpa armigera Hubner and Spodoptera litura Fabricius, in choice and no choice tests. We quantified the effects of natal host species and post-emergence adult age on the oviposition preference of the parasitoids. H. armigera eggs were consistently preferred over S. litura eggs, regardless of the natal host and adult age. When only S. litura eggs were available as hosts, they were parasitized at statistically similar rates to H. armigera eggs (average of 17 +/- 2.7 vs. 13 +/- 3.0, H. armigera to S. litura). The adult lifespan and lifetime fecundity of T. pretiosum were variable but were affected by natal host species and/or host species to which they were exposed. Mean lifespan and fecundity of parasitoids that had developed in H. armigera eggs and were exposed to H. armigera eggs for oviposition were 13.9 +/- 1.8 days and 98.7 +/- 11.0 adult offspring. By contrast, those that developed in S. litura eggs and were exposed to S. litura eggs for oviposition lived for 7 +/- 0.9 days and produced 53.8 +/- 8.0 adult offspring. The ovigeny index (OI) was significantly lower in the parasitoids exposed to H. armigera eggs than in those exposed to S. litura eggs, regardless of the natal host, indicating that H. armigera eggs sustain the adult parasitoids better than S. litura eggs. These results are used to predict parasitoid behavior in the field when both hosts are available. (c) 2006 Elsevier Inc. All rights reserved.

Comparative analysis of the relationship between phenological phenomena and meteorological indicators based on insect and plant monitoring

Climate change is one of the biggest environmental problems of the 21st century. The most sensitive indicators of the effects of the climatic changes are phenological processes of the biota. The effects of climate change which were observed the earliest are the remarkable changes in the phenology (i.e. the timing of the phenophases) of the plants and animals, which have been systematically monitored later. In our research we searched for the answer: which meteorological factors show the strongest statistical relationships with phenological phenomena based on some chosen plant and insect species (in case of which large phenological databases are available). Our study was based on two large databases: one of them is the Lepidoptera database of the Hungarian Plant Protection and Forestry Light Trap Network, the other one is the Geophytes Phenology Database of the Botanical Garden of Eötvös Loránd University. In the case of butterflies, statistically defined phenological dates were determined based on the daily collection data, while in the case of plants, observation data on blooming were available. The same meteorological indicators were applied for both groups in our study. On the basis of the data series, analyses of correlation were carried out and a new indicator, the so-called G index was introduced, summing up the number of correlations which were found to be significant on the different levels of significance. In our present study we compare the significant meteorological factors and analyse the differences based on the correlation data on plants and butterflies. Data on butterflies are much more varied regarding the effectiveness of the meteorological factors.

The possibilities of biodiversity monitoring based on Hungarian Light Trap Networks

Our method is presented with displaying time series, consisting of the daily amount of precipitation of 100 years, which has meant a separate challenge, as the precipitation data shows significant deviations. By nowadays, mankind has changed its environment to such an extent that it has a significant effect on other species as well. The Lepidoptera data series of the National Plant Protection and Forestry Light Trap Network can be used to justify this. This network has a national coverage, a large number of collected Lepidoptera, and an available, long data series of several years. For obtaining information from these data, the setting up of an easy to manage database is necessary. Furthermore, it is important to represent our data and our results in an easily analysable and expressive way. In this article the setting up of the database is introduced, together with the presentation of a three dimensional visualization method, which depicts the long-range and seasonal changes together.

Comparative reproductive biology of two Florida pawpaws Asimina reticulata Chapman and Asimina tetramera Small

I investigated the phenology and breeding systems of two Florida endemic pawpaws, Asimina reticulata, widespread in peninsular Florida, and A. tetramera, a federally endangered species limited to two counties on the Atlantic Coastal Ridge. The purpose of this study was to determine if differences contribute to the rarity of Asimina tetramera compared with A. reticulata. The study was conducted in sand pine scrub sites with the largest populations of A. tetramera in the two counties. Flowering seasons differ for the two species. Both species are hermaphroditic and strongly protogynous. Pollination experiments show that neither species is autogamous and the primary breeding mechanism is outcrossing, although low levels of geitonogamous pollination occur in mature scrub habitats. High levels of inbreeding depression were noted in both species at both sites but inbreeding depression was relaxed the first year post-fire. Fruit set in mature habitats may be pollinator limited. ^ I studied insects associated with the flowers in sand pine scrub habitat in southeastern Florida from 1994–1996. The most commonly represented orders were Coleoptera (25 spp.), Lepidoptera. (3 spp.) and Hymenoptera. (3 spp.). All Coleoptera. were flower visitors; one species, Euphoria sepulchralis (Fabricius)(Scarabeaidae), visited flowers of the two Asimina species at both sites. Eurytides marcellus (Cramer) (Lepidoptera: Papilionidae) eggs and larvae were observed on both species of Asimina during each year of the study. ^ Resource management techniques were applied to a mature sand pine scrub community in Jonathan Dickinson State Park in southeastern Florida for the management of Asimina tetramera. Manipulations conducted in 1996 included combinations of fire and mechanical treatments. I measured effects of these treatments on flowering and fruit set on A. tetramera and found cutting and burning was most effective in increasing flowering, followed by burning. Mechanical cutting and mulching had no significant effect. ^

Endocrine regulation of dynamic communication signals in gymnotiform fish

Communication signals are shaped by the opposing selection pressures imposed by predators and mates. A dynamic signal might serve as an adaptive compromise between an inconspicuous signal that evades predators and an extravagant signal preferred by females. Such a signal has been described in the gymnotiform electric fish, Brachyhypopomus gauderio, which produces a sexually dimorphic electric organ discharge (EOD). The EOD varies on a circadian rhythm and in response to social cues. This signal plasticity is mediated by the slow action of androgens and rapid action of melanocortins. My dissertation research tested the hypotheses that (1) signal plasticity is related to sociality levels in gymnotiform species, and (2) differences in signal plasticity are regulated by differential sensitivity to androgen and melanocortin hormones. To assess the breadth of dynamic signaling within the order Gymnotiformes, I sampled 13 species from the five gymnotiform families. I recorded EODs to observe spontaneous signal oscillations after which I injected melanocortin hormones, saline control, or presented the fish with a conspecific. I showed that through the co-option of the ancient melanocortin pathway, gymnotiforms dynamically regulate EOD amplitude, spectral frequency, both, or neither. To investigate whether observed EOD plasticities are related to species-specific sociality I tested four species; two territorial, highly aggressive species, Gymnotus carapo and Apteronotus leptorhynchus, a highly gregarious species, Eigenmannia cf. virescens , and an intermediate short-lived species with a fluid social system, Brachyhypopomus gauderio. I examined the relationship between the androgens testosterone and 11-ketotestosterone, the melanocortin α-MSH, and their roles in regulating EOD waveform. I implanted all fish with androgen and blank silicone implants, and injected with α-MSH before and at the peak of implant effect. I found that waveforms of the most territorial and aggressive species were insensitive to hormone treatments; maintaining a static, stereotyped signal that preserves encoding of individual identity. Species with a fluid social system were most responsive to hormone treatments, exhibiting signals that reflect immediate condition and reproductive state. In conclusion, variation in gymnotiform signal plasticity is hormonally regulated and seems to reflect species-specific sociality.

Multi-proxy records of Two-Yurts Lake

Within the scope of Russian-German palaeoenvironmental research, Two-Yurts Lake (TYL, Dvuh-Yurtochnoe in Russian) was chosen as the main scientific target area to decipher Holocene climate variability on Kamchatka. The 5x2 km large and 26 m deep lake is of proglacial origin and situated on the eastern flank of Sredinny Ridge at the northwestern end of the Central Kamchatka Valley, outside the direct influence of active volcanism. Here, we present results of a multi-proxy study on sediment cores, spanning about the last 7000 years. The general tenor of the TYL record is an increase in continentality and winter snow cover in conjunction with a decrease in temperature, humidity, and biological productivity after 5000-4500 cal yrs BP, inferred from pollen and diatom data and the isotopic composition of organic carbon. The TYL proxy data also show that the late Holocene was punctuated by two colder spells, roughly between 4500 and 3500 cal yrs BP and between 1000 and 200 cal yrs BP, as local expressions of the Neoglacial and Little Ice Age, respectively. These environmental changes can be regarded as direct and indirect responses to climate change, as also demonstrated by other records in the regional terrestrial and marine realm. Long-term climate deterioration was driven by decreasing insolation, while the short-term climate excursions are best explained by local climatic processes. The latter affect the configuration of atmospheric pressure systems that control the sources as well as the temperature and moisture of air masses reaching Kamchatka.

Purificação, caracterização e atividade bioinseticida de um inibidor de tripsina de sementes de Crotalaria pallida

A proteinaceous trypsin inhibitor was purified from Crotalaria pallida seeds by ammonium sulphate fractionation, affinity chromatography on immobilized Trypsin-Sepharose and TCA precipitation. The trypsin inhibitor, named ITC, had Mr of 32.5 kDa by SDS-PAGE and was composed by two subunits with 27.7 and 5.6 kDa linked by disulphide bridges, a typical characteristic of Kunitz-Inhibitor family. ITC was stable until 50°C, and at 100°C its residual activity was of about 60%. Also, ITC was stable at pHs 2 to 12. The inhibition of trypsin by ITC was non-competitive, with a Ki of 8,8 x 10-7M. ITC inhibits weakly other serine proteinases such as chymotrypsin and elastase. The inhibition of papain (44% of inhibition), a cysteine proteinase was an indicative of the bi-functionality of ITC. In vitro assays against digestive proteinases from several Lepdoptera, Diptera and Coleoptera pests were made. ITC inhibited in 100% digestive enzymes of Ceratitis capitata (fruit fly), Spodoptera frugiperda and Alabama argillacea, the last one being a cotton pest. It also inhibited in 74.4% Callosobruchus maculatus (bean weevil) digestive enzymes, a Coleoptera pest. ITC, when added in artificial diet models, affected weakly the development of C. capitata larvae and it had a WD50 of 2.65% to C. maculatus larvae

Purificação, caracterização e análise da atividade bioinseticida, de um inibidor de tripsina em sementes de catanduva (Piptadenia moniliformis)

One Kunitz-type trypsin inhibitors (PmTI) was purified from Piptadenia moniliformis seeds, a tree of the sub-family Mimosoideae, by TCA precipitation, affinity chromatography on immobilized trypsin-Sepharose, DEAE cellulose (ion exchange) and Superose 12 (molecular exclusion) column FPLC/AKTA. The inhibitor has Mr of 25 kDa by SDS-PAGE and chromatography molecular exclusion. The N-terminal sequence of this inhibitor showed high homology with other family Kunitz inhibitors. This also stable variations in temperature and pH and showed a small decrease in its activity when incubated with DDT in the concentration of 100mM for 120 minutes. The inhibition of trypsin by PmTI was competitive, with Ki of 1.57 x10-11 M. The activity of trypsin was effectively inhibited by percentage of inhibition of 100%, among enzymes tested, was not detected inhibition for the bromelain, was weak inhibitor of pancreatic elastase (3.17% of inhibition) and inhibited by 76.42% elastase of neutrophils, and inhibited in a moderate, chymotrypsin and papain with percentage of inhibition of 42.96% and 23.10% respectively. In vitro assays against digestive proteinases from Lepidoptera, Diptera and Coleoptera pests were carried out. Several degrees of inhibition were found. For Anthonomus grandis and Ceratitis capitata the inhibition was 89.93% and 70.52%, respectively, and the enzymes of Zabrotes subfasciatus and Callosobruchus maculatus were inhibited by 5.96% and 9.41%, respectively, and the enzymes of Plodia. interpunctella and Castnia licus were inhibited by 59.94% and 23.67, respectively. In vivo assays, was observed reduction in the development of larvae in 4rd instar of C. capitata, when PmTI was added to the artificial diet, getting WD50 and LD50 of 0.30% and 0.33%, respectively. These results suggest that this inhibitor could be a strong candidate to plant management programs cross transgenic

Microlepidoptera of Illinois hill prairies

Among insects, which are the most diverse eukaryotic group on earth, Lepidoptera is one of four enormously diverse orders, with approximately 10,000 described species in North America. Within the order, Nearctic “microlepidoptera,” which represent an overwhelmingly large percentage of diversity within the order, remain poorly known despite their ecological importance in many plant communities. In this thesis, I undertook several studies of microlepidoptera diversity in a natural community type (hill prairie) and a managed community type (biofuel feedstock). In two Illinois hill prairies differing in size, latitude, and plant composition, alpha diversity of Pyraloidea and Tortricidae was similar, but the prairies were found to support different sets of species of these moth groups. It is concluded that the similarity in alpha diversity occurs because the larger prairie supports primarily a complement of moth species that feed as larvae on prairie plants (especially species of Asteraceae and Fabaceae), whereas the moths collected in the small prairie represent relatively few prairie-associated species, plus a large component of species that feed as larvae on deciduous trees that surround the prairie. This agrees with the finding of high beta diversity of moths between the sites, which reflects a high level of larval hostplant specificity in most species of Pyraloidea and Tortricidae. Based on published information plus observations made on microlepidoptera collected during the course of this study, 31 families of basal microlepidoptera were reviewed with an aim toward evaluating the likelihood of their including species that are dependent on tallgrass prairie. Of these families, 12 were evaluated as possible, and two as likely or certain, to include prairie-dependent species. In a comparison of moth diversity in light-trap samples from corn, miscanthus, switchgrass, and native prairie, alpha diversity was highest in prairie and was higher in switchgrass than in the other two biofuel crops. Moth species complements generally were similar among the biofuel crops, and prairie shared higher species complementarity with switchgrass than with corn or miscanthus. These findings suggest that large-scale conversion of land to biofuel crops may, to a substantial degree, detrimentally affect arthropod biodiversity, with a resulting loss of valuable arthropod-derived ecosystem services both within the cropping systems and in the surrounding landscape. During the course of this study, rearing efforts yielded two species of moths of the family Gelechiidae, both of which are monophagous leaf feeders on leadplant, Amorpha canescens (Fabaceae). Because these moths are restricted to tallgrass prairie, they are likely to be of interest to conservation biologists. In the interest of naming the moths to facilitate communication regarding them, and to augment our taxonomic knowledge of their respective genera, the moths are described, and diagnoses are provided to differentiate them from similar, related species.