The effect of parasitization by Trichogramma australicum on Helicoverpa armigera host eggs and embryos

Histological investigations of the pathology of Helicoverpa armigera (Hiibner) eggs after attack by the egg parasitoid, Trichogramma australicum (Girault), indicate that the developing embryo is immediately killed by envenomation. Soon afterward the histological staining characteristics of parasitized host embryos change and the embryonic germ band dissociates into a mass of individual rounded cells. Hosts attacked by females sterilized by gamma-irradiation showed the same pathological effects as normally parasitized hosts, indicating that host degeneration is due to female venom rather than factors derived from the parasitoid embryo or larva. Cell death also occurred in older host embryos although tissue breakdown was delayed. These findings have allowed us to determine not just that the host dies but what happens to the cells and tissues, i.e., their physical appearance, the time course of their degeneration, and that the process is retarded in older hosts. These processes can possibly be emulated in artificial diets. (C) 2003 Elsevier Inc. All rights reserved.

Isolation and characterization of microsatellite loci from Helicoverpa armigera Hübner (Lepidoptera: Noctuidae)

Five microsatellite loci are presented for Helicoverpa armigera. These microsatellite loci were obtained through the construction of enriched libraries, overcoming previously reported difficulties with obtaining microsatellites from H. armigera and other Lepidoptera due to the low frequency of microsatellites in their genomes. The description of a further five microsatellite loci for H. armigera makes microsatellite based population genetics studies feasible.

Isolation and characterization of microsatellite loci from Acacia nilotica ssp. indica (Mimosaceae)

Five microsatellite loci are presented for prickly acacia, Acacia nilotica ssp. indica (Benth.) Brenan, an introduced weed of national significance in Australia. These microsatellite loci were obtained through the construction of an enriched library and their use will enable us to determine the genetic origin and extent of genetic diversity of this weed in Australia.

Isolation and characterization of microsatellite loci from Chiasmia assimilis (Warren, 1899) (Lepidoptera: Geometridae)

Twelve microsatellite loci are presented for the biological control agent Chiasmia assimilis (Warren, 1899). These microsatellite loci were obtained through the construction of an enriched library, overcoming previous reported difficulties with obtaining microsatellites from other Lepidoptera due to the low frequency of microsatellites in their genomes.

Introduced mammals in Australian rangelands: Future threats and the role of monitoring programmes in management strategies

In the present paper, we have provided an initial assessment of the current and future threats to biodiversity posed by introduced mammals (predators and herbivores) inhabiting the Australian rangelands, exploring trends in populations and options for management. Notably, rabbits have declined in recent years in the wake of rabbit haemorrhagic disease, populations of feral camels have increased dramatically and foxes appear to have moved northwards, thereby threatening native fauna within an expanded range. Following on, we developed a framework for monitoring the impacts of introduced mammals in the Australian rangelands. In doing so, we considered the key issues that needed to be considered in designing a monitoring programme for this purpose and critically evaluated the role of monitoring in pest animal management. Finally we have provided a brief inventory of current best-practice methods of estimating the abundance of introduced mammal populations in the Australian rangelands with some comments on new approaches and their potential applications.

Induction and transmission of Bacillus thuringiensis tolerance in the flour moth Ephestia kuehniella

The use of Bacillus thuringiensis (Bt) endotoxins to control insect vectors of human diseases and agricultural pests is threatened by the possible evolution of resistance in major pest species. In addition to high levels of resistance produced by receptor insensitivity (5, 16, 17), several cases of tolerance to low to medium levels of toxin have been reported in laboratory colonies of lepidopteran species (3, 18). Because the molecular basis of some of these cases of tolerance to the toxin are not known, we explored alternative mechanisms. Here, we present evidence that tolerance to a Bt formulation in a laboratory colony of the flour moth Ephestia kuehniella can be induced by preexposure to a low concentration of the Bt formulation and that the tolerance correlates with an elevated immune response. The data also indicate that both immune induction and Bt tolerance can be transmitted to offspring by a maternal effect and that their magnitudes are determined by more than one gene.

Discrepancy between antennal and behavioural responses for enantiomers of -pinene: electrophysiology and behavior of Helicoverpa armigera (Lepidoptera)

The ability of adult cotton bollworm, Helicoverpa armigera (Hubner), to distinguish and respond to enantiomers of alpha-pinene was investigated with electrophysiological and behavioral methods. Electroantennogram recordings using mixtures of the enantiomers at saturating dose levels, and single unit electrophysiology, indicated that the two forms were detected by the same receptor neurons. The relative size of the electroantennogram response was higher for the (-) compared to the (+) form, indicating greater affinity for the (-) form at the level of the dendrites. Behavioral assays investigated the ability of moths to discriminate between, and respond to the (+) and (-) forms of alpha-pinene. Moths with no odor conditioning showed an innate preference for (+)-alpha-pinene. This preference displayed by naive moths was not significantly different from the preferences of moths conditioned on (+)-alpha-pinene. However, we found a significant difference in preference between moths conditioned on the (-) enantiomer compared to naive moths and moths conditioned on (+)-alpha-pinene, showing that learning plays an important role in the behavioral response. Moths are less able to distinguish between enantiomers of alpha-pinene than different odors (e.g., phenylacetaldehyde versus (-)-alpha-pinene) in learning experiments. The relevance of receptor discrimination of enantiomers and learning ability of the moths in host plant choice is discussed.

Biology of the leech Actinobdella inequiannulata Moore, 1901 (Annelida: Hirudinea: Rhynchobdellida: Glossiphoniidae), parasitic on the white sucker, Catostomus commersoni Lacepede, 1803, and the longnose sucker, Catostomus catostomus Forster, 1773, in Algonquin Provincial Park, Ontario, Canada

Actinobdella inequiannulata was found on the white sucker. Catostomus commersoni, and less frequently on the longnose sucker, Catostomus catostomus, in Algonquin Provincial Park, Ontario, Canada. Catostomus commersoni parasitized with Act. inequiannulata was collected from July to October 1973 and May to October 1974. In May and October, less than 3% of the fish carried leeches. In July, 80% of the fish were parasitized with an average of 1.5 leeches/fish. Observations on leech weight suggest that young leeches attach to fish from May to September, some mature in July, and a second generation of leeches reparasitize the fish in August and September. The mean size of leeches on suckers increased from May until July, after which the size remained relatively constant. Leeches produced characteristic lesions on the opercula of suckers. Fully developed lesions on fish opercula produced by aggregated leeches had varying amounts of central erosion, extravasation, dermal and epidermal hyperplasia, and necrosis.

Discrimination among host plants (Leucaena species and accessions) by the psyllid pest Heteropsylla cubana and implications for understanding resistance

1 The herbivorous bug Heteropsylla cubana Crawford (Homoptera: Psyllidae) is a pest of the cattle fodder crop Leucaena (Leguminosae: Mimosoideae). The interaction between the psyllid and three varieties of its Leucaena host plant was investigated in relation to the apparent resistance of some Leucaena varieties (Leucaena leucocephala, Leucaena pallida and their hybrids) to attack. 2 Field trials demonstrated that adult psyllids distinguished among the different varieties of Leucaena over a distance, and were attracted to L. leucocephala in significantly higher numbers than to L. pallida or to the hybrid. Pesticide treatment increased the attractiveness of Leucaena plants, even of those deemed to be psyllid resistant. Numbers of psyllid eggs and nymphs, sampled in the field, reflect the arrival rates of adults at the three plant varieties. 3 Wavelength reflectance data of the three Leucaena varieties were not significantly different from one another, suggesting that psyllids cannot discriminate among the three plants using brightness or wavelength cues. There was a differential release of caryophyllene among the three varieties. Release of caryophyllene in L. leucocephala and the hybrid appeared to be influenced by environmental conditions. 4 Experiments demonstrated that caryophyllene (at least on its own) did not influence the behaviour of leucaena psyllids in relation to leucaena plants. 5 The results suggest that host plant volatiles cannot be dismissed as significant in the interaction between the leucaena psyllid and its Leucaena host plants. Further avenues for investigation are recommended and these are related to novel ways of understanding resistance in insect plant inter-relationships.

Isolation and characterization of microsatellite loci from mother-of-millions, Bryophyllum delagoense (Crassulaceae), and its hybrid with Bryophyllum daigremontianum, 'Houghton's hybrid'

Nine microsatellite loci were developed, and are transferable, across the Madagascan succulents Bryophyllum daigremontianum, Bryophyllum delagoense (mother-of-millions) and their horticultural hybrid (Houghton's), from enriched libraries of the later two species. For B. delagoense, a tetraploid, three to 13 alleles per locus were found for native Madagascan (H-O = 0.4-1.0), and one to nine in invasive Australian (H-O = 0.0-1.0) samples. In addition for 91 Australian samples, only five multilocus genotypes were found (95% of individuals were of two genotypes), suggesting extensive clonality in its introduced range. These loci will be used to examine genetic diversity, hybrid origin and mating system in natural and introduced populations.

Method for the generation and cultivation of functional three-dimensional mammary constructs without exogenous extracellular matrix

During puberty, pregnancy, lactation and postlactation, breast tissue undergoes extensive remodelling and the disruption of these events can lead to cancer. In vitro studies of mammary tissue and its malignant transformation regularly employ mammary epithelial cells cultivated on matrigel or floating collagen rafts. In these cultures, mammary epithelial cells assemble into three-dimensional structures resembling in vivo acini. We present a novel technique for generating functional mammary constructs without the use of matrix substitutes.

Increasing the efficacy of Judas goats by sterilisation and pregnancy termination

The use of Judas goats to locate remnant animals is a potentially powerful tool for enhancing goat-eradication efforts, which are especially important to island conservation. However, current Judas goat methodology falls short of its potential efficacy. Female Judas goats are often pregnant at the time of deployment or become impregnated in the field; pregnant females leave associated goats to give birth, causing downtime of Judas goat operations. Further, male Judas goats may inseminate remnant females. Sterilising Judas goats prior to deployment removes these inefficiencies. Here, we describe two methods (epididymectomy for males and tubal occlusion for females) that sterilise Judas goats while still maintaining sexual motivation and other behaviours associated with intact animals. These surgeries are straightforward, time efficient, and may be conducted in the field by staff with minimal training. Given the widespread and deleterious impacts of non-native herbivores to ecosystems and the importance of Judas operations in detecting animals at low densities, sterilisation and termination of pregnancy should be applied routinely in Judas goat (and possibly other species) programs to increase the efficacy of low-density control operations and eradication campaigns.

Forecasting Helicoverpa populations in Australia: A comparison of regression based models and a bio-climatic based modeling approach

Long-term forecasts of pest pressure are central to the effective management of many agricultural insect pests. In the eastern cropping regions of Australia, serious infestations of Helicoverpa punctigera (Wallengren) and H. armigera (Hübner)(Lepidoptera: Noctuidae) are experienced annually. Regression analyses of a long series of light-trap catches of adult moths were used to describe the seasonal dynamics of both species. The size of the spring generation in eastern cropping zones could be related to rainfall in putative source areas in inland Australia. Subsequent generations could be related to the abundance of various crops in agricultural areas, rainfall and the magnitude of the spring population peak. As rainfall figured prominently as a predictor variable, and can itself be predicted using the Southern Oscillation Index (SOI), trap catches were also related to this variable. The geographic distribution of each species was modelled in relation to climate and CLIMEX was used to predict temporal variation in abundance at given putative source sites in inland Australia using historical meteorological data. These predictions were then correlated with subsequent pest abundance data in a major cropping region. The regression-based and bioclimatic-based approaches to predicting pest abundance are compared and their utility in predicting and interpreting pest dynamics are discussed.

Generation and maturation of dendritic cells for clinical application under serum-free conditions

Monocyte-derived dendritic cells (MoDCs) in clinical use for cancer immunotherapy are ideally generated in serum-free medium (SFM) with inclusion of a suitable maturation factor toward the end of the incubation period. Three good manfacturing practice (GMP) grade SFMs (AIM-V, X-VIVO 15, and X-VIVO 20) were compared with RPMI-1640, supplemented with 10% fetal bovine serum or 10% human serum. DCs generated for 7 days in SFM were less mature and secreted less interleukin (IL) 12p70 and IL-10 than DCs generated in 10% serum. DC yield was comparable in SFMs, and a greater proportion of cells was viable after maturation. Toll-like receptor (TLR) ligands were compared for their ability to induce cytokine secretion under serum-free conditions in the presence of interferon (IFN) gamma. With the exception of Poly I:C, TLR ligands stimulated high levels of IL-10 secretion. High levels of IL-12p70 were induced by two TLR4-mediated stimuli, lipopolysaccharide and Ribomunyl, a clinical-grade bacterial extract. When T-cell responses were compared in allogeneic mixed leukocyte reaction, DCs stimulated with Ribomunyl induced higher levels of IFN gamma than DCs stimulated with the cytokine cocktail: tumor necrosis factor-alpha, IL-1 beta, IL-6, and prostaglandin E-2. In the presence of IL-10 neutralizing antibodies, DC IL-12p70 production and T-cell IFN gamma were increased in vitro. Similarly, DCs stimulated with Ribomunyl, IFN gamma, and anti-IL-10 induced high levels of tetanus toxoid-specific T-cell proliferation and IFN gamma secretion. Thus, MoDCs generated ill SFM efficiently stimulate T-cell IFN gamma production after maturation in the presence of a clinical-grade TLR4 agonist and IL-10 neutralization.