Progression of phosphine resistance in susceptible Tribolium castaneum (Herbst) populations under different immigration regimes and selection pressures

Insecticide resistance is an escalating global issue for a wide variety of agriculturally important pests. The genetic basis and biochemical mechanisms of resistance are well characterized in some systems, but little is known about the ecological aspects of insecticide resistance. We therefore designed a laboratory experiment to quantify the progression of phosphine resistance in Tribolium castaneum populations subject to different immigration regimes and selection pressures. Mated resistant females were added to originally susceptible populations under two distinct migration rates, and in addition, half of the populations in each migration treatment were exposed to selection pressures from phosphine fumigation. The progression of phosphine resistance was assessed by screening beetles for the resistance allele at rph2. Phosphine resistance increased slowly in the low migration treatment and in the absence of selection, as expected. But at the higher migration rate, the increase in frequency of the resistance allele was lower than predicted. These outcomes result from the high levels of polyandry known in T. castaneum females in the laboratory, because most of the Generation 1 offspring (86%) were heterozygous for the rph2 allele, probably because resistant immigrant females mated again on arrival. Phosphine resistance was not fixed by fumigation as predicted, perhaps because susceptible gametes and eggs survived fumigation within resistant females. In terms of phosphine resistance progression in populations exposed to selection, the effect of fumigation negated the difference in migration rates. These results demonstrate how species-specific traits relating to the mating system may shape the progression of insecticide resistance within populations, and they have broad implications for the management of phosphine resistance in T. castaneum in the field. ​We specify and discuss how these mating system attributes need to be accounted for when developing guidelines for resistance management

Phosphine fumigation of cool grain

The biosecurity problem addressed was the need to understand and evaluate phosphine fumigation of cool grain (i.e. 20°C or less) as a means of controlling resistant biotypes of insect pests of stored grain which are major EPPs threatening the grain industry. The benefits of cooling and phosphine fumigation are that cooling preserves grain quality and reduces insect population growth, and phosphine kills insects and has a residue free status in all major markets. The research objectives were to: - conduct laboratory experiments on phosphine efficacy against resistant insects in cool grain, and determine times to population extinction. - conduct laboratory experiments on phosphine sorption in cool grain and quantify. - complete fumigation trials in three states (Queensland, WA and NSW) on cool grain stored insealed farm silos. - make recommendations for industry on effective phosphine fumigation of cool grain. Phosphine is used by growers and other stakeholders in the grain industry to meet domesticand international demands for insect-free grain. The project aim was to generate new information on the performance of phosphine fumigation of cool grain relevant to resistant biotypes. Effective control of resistant biotypes using phosphine to fumigate cool grain will benefit growers and other sectors of the grain industry, needing to fumigate grain in the cooler months of the year, or grain that has been cooled using aeration

Resistance management and the ecology of Rhyzopertha dominica (F.) and Tribolium castaneum (Herbst) in subtropical Australia

Subtropical Australia is a demonstrated hotspot for phosphine resistance in stored product pests. Rhyzopertha dominica and Tribolium castaneum are common pests of stored grain in this region and management of these pests is increasingly impeded through the spread of resistance to phosphine, the most desirable control method. A number of field-oriented studies were conducted from the 1970’s to 1990’s to understand the ecology of these pests in subtropical Australia, including seasonal abundance, flight and population growth in stored grain. To manage the evolution and spread of resistance we require an understanding of movement of these beetles among foci of infestation. This paper presents preliminary analyses of two aspects of new research on these species: (1) a trapping program using pheromones to investigate beetle numbers in spatial and temporal contexts, and (2) characterisation of beetles leaving infested farm silos. Adults of both species were trapped throughout the year with the lowest numbers corresponding to the coldest part of the year. The coldest trapping period had mean maximum and minimum temperatures of 21.1 and 3.5°C respectively. Trapping also revealed distinct differences between the two species, both in terms of numbers caught and where they were caught. In general, more R. dominica were caught than T. castaneum, similar numbers of R. dominica were caught near farm silos and in paddocks at least 1 km from the nearest silo, and more T. castaneum were caught near silos than in paddocks. Individual adults intercepted flying from farm silos are being characterised in the laboratory, and results to date show that these adults are long-lived, the females have mated before emigrating and are highly fecund. By undertaking research of the type summarised here we aim to develop an understanding of how these two species interact with their environment and how these interactions influence resistance development. Keywords: Ecology, Rhyzopertha dominica, Tribolium castaneum, Australi

Policy, innovation and cost reduction in UK offshore wind

Offshore wind in the UK has been a remarkable green growth success story. The price of offshore wind is less than a third of what it was a decade ago (Figure 1). In UK conditions, it is now cost competitive with new fossil fuel generation1. Indeed, if electricity prices return to pre-Covid levels, the Government would no longer be subsidising new offshore wind; HM Treasury will be earning revenue instead2. In parallel, the industry has grown and matured to a point where oil and gas companies are clamouring to enter the market and pension funds are comfortable in investing billions of pounds into construction

Prevalence and potential fitness cost of weak phosphine resistance in Tribolium castaneum (Herbst) in eastern Australia

The prevalence of resistance to phosphine in the rust-red flour beetle, Tribolium castaneum, from eastern Australia was investigated, as well as the potential fitness cost of this type of resistance. Discriminating dose tests on 115 population samples collected from farms from 2006 to 2010 showed that populations containing insects with the weakly resistant phenotype are common in eastern Australia (65.2 of samples), although the frequency of resistant phenotypes within samples was typically low (median of 2.3). The population cage approach was used to investigate the possibility that carrying the alleles for weak resistance incurs a fitness cost. Hybridized populations were initiated using a resistant strain and either of two different susceptible strains. There was no evidence of a fitness cost based on the frequency of susceptible phenotypes in hybridized populations that were reared for seven generations without exposure to phosphine. This suggests that resistant alleles will tend to persist in field populations that have undergone selection even if selection pressure is removed. The prevalence of resistance is a warning that this species has been subject to considerable selection pressure and that effective resistance management practices are needed to address this problem. The resistance prevalence data also provide a basis against which to measure management success

Prevalence and potential fitness cost of weak phosphine resistance in Tribolium castaneum (Herbst) in eastern Australia

The prevalence of resistance to phosphine in the rust-red flour beetle, Tribolium castaneum, from eastern Australia was investigated, as well as the potential fitness cost of this type of resistance. Discriminating dose tests on 115 population samples collected from farms from 2006 to 2010 showed that populations containing insects with the weakly resistant phenotype are common in eastern Australia (65.2 of samples), although the frequency of resistant phenotypes within samples was typically low (median of 2.3). The population cage approach was used to investigate the possibility that carrying the alleles for weak resistance incurs a fitness cost. Hybridized populations were initiated using a resistant strain and either of two different susceptible strains. There was no evidence of a fitness cost based on the frequency of susceptible phenotypes in hybridized populations that were reared for seven generations without exposure to phosphine. This suggests that resistant alleles will tend to persist in field populations that have undergone selection even if selection pressure is removed. The prevalence of resistance is a warning that this species has been subject to considerable selection pressure and that effective resistance management practices are needed to address this problem. The resistance prevalence data also provide a basis against which to measure management success

Gene introgression in assessing fitness costs associated with phosphine resistance in the rusty grain beetle

The current study investigates the fitness cost associated with phosphine resistance in the rusty grain beetle, Cryptolestes ferrugineus (Stephens), a problematic pest in the stored commodities that has developed strong resistance to fumigant phosphine. Three characterised insect strains: the susceptible (Ref-S), the strongly resistant (Ref-R), the introgressed resistant (Intro-R) and a segregating population (F25) derived from crossing the Ref-S and Ref-R strains were used in this study. Intro-R was developed by introgressing two phosphine resistance genes, cf_rph1 and cf_rph2 into Ref-S, aimed to reduce the influence of background genetic factors. Intro-R exhibited 592 × resistance to phosphine and homozygous for strong resistance allele, cf_rph2 (L73N). Two key fitness cost criteria, developmental time and fecundity, were assessed under optimal and suboptimal conditions (less favourable diet and low temperature). There was no significant difference in developmental time and fecundity between Ref-S and either Intro-R strain or F25 under optimal conditions. When challenged with a less favourable diet, cracked wheat + cracked sorghum (CW + CS), or exposed to a low temperature (22 °C), both Intro-R and Ref-S had similar developmental time and total numbers of F1 progeny, confirming the absence of significant fitness effects expressed in these conditions. Therefore, we conclude that strongly phosphine resistant C. ferrugineus are unlikely to incur potential fitness costs. This finding will have implications towards developing strategies to manage this pest

Potential for using pheromone trapping and molecular screening in phosphine resistance research: Presentation

Phosphine resistance monitoring typically involves bioassays of beetles from population samples collected from grain storage facilities. Insects are classified into susceptible or resistant phenotypes based on mortality or survival at one or more discriminating doses. Although valuable, phenotype testing has several drawbacks. First, phenotype testing needs live insects, and considerable effort is required to collect and maintain them before testing. Second, population samples may contain multiple genotypes expressing different levels of resistance that may not be distinguishable using discriminating dose bioassays. Third, collections are likely to be focussed around grain storages to maximise sampling success. Recent research shows that several key pests are actively dispersing through flight. The availability of commercial pheromone lures and recent advances in molecular screening provide an opportunity to provide information on resistance gene frequencies more broadly across the landscape. This approach is proving to be a valuable adjunct to traditional resistance testing in Australia.Phosphine resistance monitoring typically involves bioassays of beetles from population samples collected from grain storage facilities. Insects are classified into susceptible or resistant phenotypes based on mortality or survival at one or more discriminating doses. Although valuable, phenotype testing has several drawbacks. First, phenotype testing needs live insects, and considerable effort is required to collect and maintain them before testing. Second, population samples may contain multiple genotypes expressing different levels of resistance that may not be distinguishable using discriminating dose bioassays. Third, collections are likely to be focussed around grain storages to maximise sampling success. Recent research shows that several key pests are actively dispersing through flight. The availability of commercial pheromone lures and recent advances in molecular screening provide an opportunity to provide information on resistance gene frequencies more broadly across the landscape. This approach is proving to be a valuable adjunct to traditional resistance testing in Australia