New tools for management of phosphine resistance

Phosphine is the primary fumigant used to protect the majority of the world' s grain and a variety of other stored commodities from insect pests. Phosphine is playing an increasingly important role in the protection of commodities for two primary reasons. Firstly, use of the alternative fumigant, methyl bromide, has been sharply curtailed and is tightly regulated due to its role in ozone depletion, and secondly, consumers are becoming increasingly intolerant of contact pesticides. Niche alternatives to phosphine exist, but they suffer from a range of factors that limit their use, including: 1) Limited commercial adoption due to expense or slow mode of action; 2) Poor efficacy due to low toxicity, rapid sorption, limited volatility or high density; 3) Public health concerns due to toxicity to handlers or nearby residents, as well as risk of explosion; 4) Poor consumer acceptance due to toxic residues or smell. These same factors limit the prospects of quickly identifying and deploying a new fumigant. Given that resistance toward phosphine is increasing among insect pests, improved monitoring and management of resistance is a priority. Knowledge of the mode of action of phosphine as well as the mechanisms of resistance may also greatly reduce the effort and expense of identifying synergists or novel replacement compounds.

Gene interactions constrain the course of evolution of phosphine resistance in the lesser grain borer, Rhyzopertha dominica

Phosphine, a widely used fumigant for the protection of stored grain from insect pests, kills organisms indirectly by inducing oxidative stress. High levels of heritable resistance to phosphine in the insect pest of stored grain, Rhyzopertha dominica have been detected in Asia, Australia and South America. In order to understand the evolution of phosphine resistance and to isolate the responsible genes, we have undertaken genetic linkage analysis of fully sensitive (QRD14), moderately resistant (QRD369) and highly resistant (QRD569) strains of R. dominica collected in Australia. We previously determined that two loci, rph1 and rph2, confer high-level resistance on strain QRD569, which was collected in 1997. We have now confirmed that rph1 is responsible for the moderate resistance of strain QRD369, which was collected in 1990, and is shared with a highly resistant strain from the same geographical region, QRD569. In contrast, rph2 by itself confers only very weak resistance, either as a heterozygote or as a homozygote and was not discovered in the field until weak resistance (probably due to rph1) had become ubiquitous. Thus, high-level resistance against phosphine has evolved via stepwise acquisition of resistance alleles, first at rph1 and thereafter at rph2. The semi-dominance of rph2 together with the synergistic interaction between rph1 and rph2 would have led to rapid selection for homozygosity. A lack of visible fitness cost associated with alleles at either locus suggests that the resistance phenotype will persist in the field.

Sampling and management of Bemisia tabaci (Genn.) biotype B in Australian cotton

Data on seasonal population abundance of Bemisia tabaci biotype B (silverleaf whitefly (SLW)) in Australian cotton fields collected over four consecutive growing seasons (2002/2003-2005/2006) were used to develop and validate a multiple-threshold-based management and sampling plan. Non-linear growth trajectories estimated from the field sampling data were used as benchmarks to classify adult SLW field populations into six density-based management zones with associated control recommendations in the context of peak flowering and open boll crop growth stages. Control options based on application of insect growth regulators (IGRs) are recommended for high-density populations (>2 adults/leaf) whereas conventional (non-IGR) products are recommended for the control of low to moderate population densities. A computerised re-sampling program was used to develop and test a binomial sampling plan. Binomial models with thresholds of T=1, 2 and 3 adults/leaf were tested using the field abundance data. A binomial plan based on a tally threshold of T=2 adults/leaf and a minimum sample of 20 leaves at nodes 3, 4 or 5 below the terminal is recommended as the most parsimonious and practical sampling protocol for Australian cotton fields. A decision support guide with management zone boundaries expressed as binomial counts and control options appropriate for various SLW density situations is presented. Appropriate use of chemical insecticides and tactics for successful field control of whiteflies are discussed.

Spatial patterns of sub-tidal seagrasses and their tissue nutrients in the Torres Strait, northern Australia: Implications for management

The distribution and nutritional profiles of sub-tidal seagrasses from the Torres Strait were surveyed and mapped across an area of 31,000 km2. Benthic sediment composition, water depth, seagrass species type and nutrients were sampled at 168 points selected in a stratified representative pattern. Eleven species of seagrass were present at 56 (33.3%) of the sample points. Halophila spinulosa, Halophila ovalis, Cymodocea serrulata and Syringodium isoetifolium were the most common species and these were nutrient profiled. Sub-tidal seagrass distribution (and associated seagrass nutrient concentrations) was generally confined to northern-central and south-western regions of the survey area (herbivores, were significantly correlated with species and with the plant component (above or below ground). For all seagrass species, the above-ground component (shoots and leaves) possessed greater nitrogen concentrations than the below-ground component (roots and rhizomes), which possessed greater starch concentrations. S. isoetifolium had the highest total nitrogen concentrations (1.40±0.05% DW). However, it also had higher fibre concentrations (38.2±0.68% DW) relative to the other four species. H. ovalis possessed the highest starch concentrations (2.76±0.12% DW) and highest digestibility (83.24±0.66% DW) as well as the lowest fibre (27.2±0.66% DW). The high relative abundance (found at 55% of the sites that had seagrass) and nutrient quality characteristics of H. ovalis make it an important source of energy to marine herbivores that forage sub-tidally in the Torres Strait. There were two regions in Torres Strait (north-central and south-western) where sub-tidal seagrass meadows were prevalent and of relatively higher nutritional value. This spatial and nutritional information can be used by local agencies to manage and to protect the ecological, economic and cultural values of the sub-tidal seagrass ecosystems and associated fisheries of the Torres Strait.

The effects of salivary gland extracts from Boophilus microplus ticks on mitogen-stimulated bovine lymphocytes

The effect of salivary gland extract (SGE) from the tick Boophilus microplus was examined in mitogen-stimulated lymphocytes in vitro. SGE was added to lymphocytes of seven cattle together with the mitogens concanavalin A (ConA), phytohaemagglutinin (PHA) and pokeweed mitogen (PWM). Semi-purified B cells from another seven cattle were stimulated with the mitogen lipopolysaccharide (LPS). PHA and ConA stimulated proliferation of lymphocytes to the same extent, but the inhibition due to SGE of Boophilus microplus on the proliferative response stimulated by PHA (39.0% ± 9.3%) was less than the inhibition of proliferative response stimulated by ConA (75.4% ± 6.9%). In contrast, SGE of B. microplus stimulated the proliferation of B cells in the presence of LPS in a dose-dependent manner. Enhanced stimulation of B cells by SGE at >4 μg in culture was greater than twice that observed when B cells were stimulated by LPS alone. SGE does not have a direct suppressive effect on bovine B cell proliferation; however, in vivo the effectiveness of B cell responses might be influenced by other immune factors, such as cytokine profiles.

Ceratocystis atrox sp. nov. associated with Phoracantha acanthocera infestations on Eucalyptus grandis in Australia

Ceratocystis spp. include important pathogens of trees as well as apparently saprophytic species. Four species have been recorded on Eucalyptus grandis in Australia, of which only one, C. pirilliformis Barnes and M.J. Wingf., is known to be pathogenic. A recent survey of pests and diseases of Eucalyptus trees in northern Queensland revealed a species of Ceratocystis associated with the tunnels made by the aggressive wood-boring insect Phoracantha acanthocera (Macleay) (Cerambicydae: Coleoptera). The aim of the present study was to identify the fungus based on morphological characteristics and comparisons of DNA sequence data for three gene regions. The fungus peripherally resembles C. fimbriata Ell. and Halst. but differs from this species most obviously by having much darker mycelium, longer ascomatal necks, segmented hyphae and an absence of aleuroconidia. Comparisons of combined sequence data confirmed that the Ceratocystis sp. from P. acanthocera represents an undescribed taxon, which is provided with the name Ceratocystis atrox sp. nov. C. atrox appears to have a close relationship with P. acanthocera, although its role in the biology of the insect is unknown and its pathogenicity has not been considered.

Optimising indoor phosphine fumigation of paddy rice bag-stacks under sheeting for control of resistant insects

Three indoor, sheeted bag-stack fumigations of paddy rice using aluminium phosphide were undertaken in Guangdong Province, southern China. We measured the effect of two types of sheeting (polyvinylchloride [PVC] or polyethylene [PE]) and two types of floor sealing (clips or fixing into a slot with a rubber pipe) on phosphine concentration and retention. The aim was to test the feasibility of retaining fumigant at a sufficient concentration for long enough to control known resistant insect pests. Each stack was pressure tested and phosphine concentrations measured daily during the fumigation. Cages of test insects in culture medium, including resistant and susceptible strains, were placed inside each stack and could be observed through the clear sheeting. Highest concentrations for the longest period were obtained in a PVC-covered stack that included a ground sheet and wall sheets sealed to the floor with rubber pipes. A similar PVC-covered stack sealed to the floor with clips instead of pipe did not retain gas as efficiently and required re-dosing. A PE-covered stack, with no ground sheet but also with wall sheets sealed to the floor with pipe, produced an acceptable fumigation. Susceptible Rhyzopertha dominica were controlled in 2 days and the most resistant strain in 15 days. Resistant Cryptolestes ferrugineus survived until day 21. The paddy was still free of insect infestation 7 months later when the bag-stack was opened to mill the rice. Pressure half-lives correlated with gas concentration and retention. Sorption appeared to be a major limiting factor, reducing potential fumigant dosage by about 50%. The trials demonstrated the feasibility of sealing bag-stacks to a standard high enough to control all known resistant strains.

Host plant resistance in grain crops and prospects for invertebrate pest management in Australia: An overview

An integrated pest management (IPM) approach that relies on an array of tactics is adopted commonly in response to problems with pesticide-based production in many agricultural systems. Host plant resistance is often used as a fundamental component of an IPM system because of the generally compatible, complementary role that pest-resistant crops play with other tactics. Recent research and development in the resistance of legumes and cereals to aphids, sorghum midge resistance, and the resistance of canola varieties to mite and insect pests have shown the prospects of host plant resistance for developing IPM strategies against invertebrate pests in Australian grain crops. Furthermore, continuing advances in biotechnology provide the opportunity of using transgenic plants to enhance host plant resistance in grains.

Spiroacetal biosynthesis in insects from diptera to hymenoptera: The Giant Ichneumon wasp Megarhyssa nortoni nortoni Cresson

The volatile components of the mandibular gland secretion generated by the Giant Ichneumon parasitoid wasp Megarhyssa nortoni nortoni Cresson are mainly spiroacetals and methyl ketones, and all have an odd number of carbon atoms. A biosynthetic scheme rationalizing the formation of these diverse components is presented. This scheme is based on the results of incorporation studies using 2H-labeled precursors and [18O]dioxygen. The key steps are postulated to be decarboxylation of β-ketoacid equivalents, β-oxidation (chain shortening), and monooxygenase-mediated hydroxylation leading to a putative ketodiol that cyclizes to spiroacetals. The generality of the role of monooxygenases in spiroacetal formation in insects is considered, and overall, a cohesive, internally consistent theory of spiroacetal generation by insects is presented, against which future hypotheses will have to be compared.

Analysing industry needs to fine tune IPM project activities

As a first step to better targeting the activities of a project for improving management of western flower thrips, Frankliniella occidentialis, (WFT) in field grown vegetable crops, we surveyed growers, consultants and other agribusiness personnel in two regions of Queensland. Using face-to-face interviews, we collected data on key pests and measures used to manage them, the importance of WFT and associated viral diseases, sources of pest management information and additional skills and knowledge needed by growers and industry. Responses were similar in the two regions. While capsicum growers in one northern Queensland district had suffered serious losses from WFT damage in 2002, in general the pest was not seen as a major problem. In cucurbit crops, the silverleaf whitefly (Bemisia tabaci biotype B) was considered the most difficult insect pest to manage. Pest control tactics were largely based on pesticides although many respondents mentioned non-chemical methods such as good farm hygiene practices, control of weed hosts and regular crop monitoring, particularly when prompted. Respondents wanted to know more about pest identification, biology and damage, spray application and the best use of insecticides. Natural enemies were mentioned infrequently. Keeping up to date with available pesticide options, availability of new chemicals and options for a district-wide approach to managing pests emerged as key issues. Growers identified agricultural distributors, consultants, Queensland Department of Primary Industries staff, other growers and their own experience as important sources of information. Field days, workshops and seminars did not rank highly. Busy vegetable growers wanted these activities to be short and relevant, and preferred to be contacted by post and facsimile rather than email. In response to these results, we are focusing on three core, interrelated project extension strategies: (i) short workshops, seminars and farm walks to provide opportunities for discussion, training and information sharing with growers and their agribusiness advisors; (ii) communication via newsletters and information leaflets; (iii) support for commercialisation of services.

Management of mould mite Tyrophagus putrescentiae (Schrank) (Acarina: Acaridae): A case study in stored animal feed

An integrated pest management (IPM) strategy was developed to manage infestations of mould mite Tyrophagus putrescentiae (Schrank) in stored animal feed, due to the increasing importance of these mites as pests of feed processing and storage facilities in Australia. This strategy involved several aspects such as limiting the moisture content of the processed feed to 12%, admixing vegetable oil to some feed (2% w/w), strict hygiene practice in and around the processing and storage facility, and rejection of infested grain at the receiving point. Additionally, seven contact insecticides and the fumigant phosphine were evaluated for their effectiveness against the mould mite to assess their potential integration into the IPM strategy. Among them, pyrethrin synergised with piperonyl butoxide, the insect growth regulator s-methoprene and a newly developed bacterium-based material spinosad controlled the mites. Moreover, the fumigant phosphine at 1 mg/litre over a six days exposure period also controlled these mites. So far, the IPM strategy, without any involvement of insecticides or fumigant has resulted in a complete eradication of the mite population in this particular case of stored animal feed.

Hidden trails: Visualizing arthropod silk

Arthropods are known to use silk for a number of different purposes including web construction, shelter building, leaf tying, construction of pupal cocoons, and as a safety line when dislodged from a substrate (Alexander, 1961; Fitzgerald, 1983; Common, 1990). Across the arthropods, silk displays a diversity of material properties and chemical constituents and is produced from glands with different evolutionary origins (Craig, 1997). Among insects, larval Lepidoptera are prolific producers of silk. Because many lepidopteran larvae are pests, an ability to interfere with silk production or, at the very least, an understanding of how silk is used, could provide new options for pest control. After testing many known fluorescent dyes, we found that Fluorescent Brightener 28 (also known as Calcofluor White M2R) (Sigma-Aldrich Pty Ltd, Sydney, NSW, Australia), an optical brightener used in the textile industry, binds to arthropod silk in a simple staining reaction, causing it to fluoresce under ultraviolet (UV) light. Such brighteners have also been used in insect gut content analysis (Schlein & Muller, 1995; Hugo et al., 2003). Here we describe the method of visualizing arthropod silk on plant surfaces, using as a model the thin, barely visible, single strands of silk produced by Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) neonates.

Plant hosts of the phytoplasmas and rickettsia-like-organisms associated with strawberry lethal yellows and green petal diseases

Candidatus Phytoplasma australiense (Ca. P. australiense) is associated with the plant diseases strawberry lethal yellows (SLY), strawberry green petal (SGP), papaya dieback (PDB), Australian grapevine yellows (AGY) and Phormium yellow leaf (PYL; New Zealand). Strawberry lethal yellows disease is also associated with a rickettsia-like-organism (RLO) or infrequently with the tomato big bud (TBB) phytoplasma, the latter being associated with a wide range of plant diseases throughout Australia. In contrast, the RLO has been identified only in association with SLY disease, and Ca. P. australiense has been detected only in a limited number of plant host species. The aim of this study was to identify plant hosts that are possible reservoirs of Ca. P. australiense and the SLY RLO. Thirty-one plant species from south-east Queensland were observed with disease between 2001 and 2003 and, of these, 18 species tested positive using phytoplasma-specific primers. The RLO was detected in diseased Jacksonia scoparia and Modiola caroliniana samples collected at Stanthorpe. The TBB phytoplasma was detected in 16 different plant species and Ca. P. australiense Australian grapevine yellows strain was detected in six species. The TBB phytoplasma was detected in plants collected at Nambour, Stanthorpe, Warwick and Brisbane. Ca. P. australiense was detected in plants collected at Nambour, Stanthorpe, Gatton and Allora. All four phytoplasmas were detected in diseased Gomphocarpus physocarpus plants collected at Toowoomba, Allora, Nambour and Gatton. These results indicated that the vector(s) of Ca. P. australiense are distributed throughout south-east Queensland and the diversity of phytoplasmas detected in G. physocarpus suggests it is a feeding source for phytoplasma insect vectors or it has a broad susceptibility to a range of phytoplasmas.

Screening selected myrtaceous tree species for production of essential oils in northern Queensland

Trials to identify alternative cropping options to Melaleuca alternifolia for northern Queensland essential oil growers were established at Dimbulah and Innot Hot Springs in 2001. Seed sources of Asteromyrtus symphyocarpa (1,8-cineole form), Eucalyptus staigeriana (citral), Melaleuca cajuputi subsp. cajuputi (trans-nerolidol), M. ericifolia (d-linalool), M. quinquenervia (trans-nerolidol and viridiflorol forms) and M. viridiflora (methyl cinnamate) with potential to produce commercial foliar oils were evaluated. Information was gathered on their adaptability, growth and oil yields over 49 months and 52 months (two harvests) from planting at Dimbulah and Innot Hot Springs, respectively. Of the species and chemotypes evaluated, M. quinquenervia showed potential for commercial production of trans-nerolidol, a compound used in perfumery. It had a very high survival rate (96%) and yields could be expected to improve dramatically from the average 100 kg/ha per harvest achieved in these trials with further research into selection of seed source, control of insect damage and breeding for genetic improvement. M. cajuputi subsp. cajuputi gave a similar performance to M. quinquenervia. The rarity of the trans-nerolidol form of this species and remoteness of its natural occurrence are impediments to further planting and research. E. staigeriana, with second harvest yields of ~600 kg/ha, performed exceptionally well on both sites but potential for development is limited by the ready availability of competitively priced E. staigeriana oil produced in South America. Survival of M. ericifolia ranged from 62% to 82% at 32 months (second harvest) at Innot Hot Springs and was deemed a failure at Dimbulah with poor growth and low survival, raising a major question about the suitability of this species for cultivation in the seasonally dry tropics. Planting of this species on a wider scale in northern Queensland cannot be recommended until more is known about factors affecting its survival. A. symphyocarpa and M. viridiflora were too slow-growing to warrant further consideration as potential oil-producing species at this time.

Agonosoma trilineatum (Heteroptera: Scutelleridae) a biological control agent of the weed bellyache bush, Jatropha gossypiifolia (Euphorbiaceae)

Bellyache bush, Jatropha gossypiifolia L., is a serious weed of northern Australia. Agonosoma trilineatum (F.) is an insect from tropical America released in Australia in 2003 as a biological control agent against bellyache bush. It feeds on seeds and has the potential to reduce seed production, thereby potentially reducing the rate of spread and recruitment. To test the host specificity of A. trilineatum, four biological responses to host plant species were determined: development of nymphs, oviposition preferences, adult feeding and frequency of mating. Development of nymphs to adults and adult feeding only occurred on three Jatropha spp. These species also supported mating and oogenesis but only J. gossypiifolia was accepted for oviposition. Mating did not occur in the presence of other plant species. The evidence indicates that there is little risk associated with the release of this insect species in Australia and probably other countries where this weed is a problem. The probability of this insect expanding its host range is low because multiple aspects of the biology would need to change simultaneously. A. trilineatum was released in Australia between 2003 and 2007. A Climex model indicated that coastal areas of Queensland and the Northern Territory would be climatically most suitable for this insect.