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Resistance of stored product insects to phosphine: an update

14 October 20198 min reading

“Monitoring is essential in phosphine use. When it comes to monitoring, monitoring of the gas concentration is not the only thing that has to be monitored. Other parameters, such as temperature etc. should be monitored in parallel, as phosphine concentration has interesting diurnal circles according to the key abiotic conditions prevailing. Distribution of the gas is also important. The traditional monitoring techniques are able to provide measurements for specific time intervals and at specific locations within the fumigated facilities. However, the measurements refer to the average concentrations, which are not always representative. Extreme dosing and absence of monitoring are key elements that are directly related with resistance development.”

Christos G. Athanassiou Laboratory of Entomology and Agricultural Zoology Department of Agriculture, Crop Production and Rural Environment University of Thessaly, Greece athanassiou@agr.uth.gr

INTRODUCTION Despite the newer developments with liquid insecticides in stored product protection, e.g. introduction of new active ingredients to the market, the fumigant phosphine remains the main insecticide that is used at the post-harvest stages of durable agricultural commodities, due to its effectiveness and the convenience in application. The development of resistance to insecticides is an evolutionary phenomenon, that has been thoroughly studied from both molecular and population genetics. It is thus a complicated phenomenon, that involves basic gene mutations and changes in chromosome structure, and is directly related with fitness cost and insect behavioral characteristics. Although the degree on which human involvement is related with the development of resistance has not been clarified in detail, there are specific indicators that can be used when the presence of resistance is present. These can be summarized in the following:

What happens to the genes if the fumigant is not applied? There are certain active ingredients that can be used with success towards this direction. Rotation of phosphine with other insecticides is the key element in resistance management. These other insecticides are referred often as “resistance breakers”, and can be applied in a carefully planned strategy on the basis of resistance mitigation practices. There are numerous paradigms, for different insecticides that are currently in use in stored product protection, that the avoidance in the use of specific active ingredients for some intervals may “reverse” resistance development through different resistance gene frequencies. This “breaker” approach has yielded good results in several parts of the world, and can be further expanded. For example, liquid insecticides, that can be effective for a long period on grains and related durable commodities have been proposed for this purpose, as their persistence on the commodity is able to lead to the drastic reduction of the fumigations that are performed for a certain period of time.

A FITNESS COST IS INVOLVED Different factors such as insect movement and mating may also impact the frequency of resistance. Considering the availability of new research tools in studying insect behavior, it is now well established that, in some cases, there is a considerable fitness cost of the resistant populations. For example, respiration may be reduced, as well as progeny production capacity, while development may be slower than that of the susceptible populations. This is directly related with frequency and occurrence of resistance, and is likely to cause a more gradual increase in resistance cases, instead of a rapid peak in the presence of the resistance genes. Still, differences in behavior may result in differential response of the resistant individuals to sampling and trapping methods, e.g. reduced movement and attraction towards a pheromone source in trapping protocols.

THERE ARE MOLECULAR MARKERS THAT CAN BE USED FOR EVALUATION The sequence information of the rph2 phosphine resistance gene has been used to develop molecular markers. This has been done in Australia, India, USA and Turkey. Although multiple alleles in DLD that are related with phosphine resistance have been recorded in Australia, there was only one allele from other countries (P45/49S). From a practical point of view, these markers can be further examined to separate weak from strong resistance, which is a key issue is some areas, as strongly resistant individuals should be treated in different way. Overall, there are certain advantages on the use of molecular/genetic markers for resistance, over the use of the classical laboratory evaluations, such as the dose-bioassay protocols (i.e. exposure of the insects at different concentrations in jars). The most important advantage is that the molecular markers can provide early detection, and thus the potential of a population to develop resistance can be seen earlier than using bioassays.

Graph: Phosphine concentration (orange line) is fluctuating during the day, affected by grain temperature (blue line) and air currents in the grain mass (monitoring using the centaur.ag technology)

THERE ARE RAPID DIAGNOSTICS AVAILABLE Apart from molecular markers that underline the presence of phosphine resistance genes, there are quick diagnostics that can provide accurate data in a very short period of time, usually hours or even minutes. These diagnostics are based on deviation of insects from normal movement and evaluate time to knockdown or immobilization. This is particularly important, as some of these diagnostics can be operated on site, e.g. by a fumigator in a flour mill prior the initiation of the fumigation. There are important advantages over the use of these diagnostics, as they are much less laborious than dose-response studies that last for days, and require specialized laboratories. It remains unclear, however, if these diagnostics can identify populations that are strongly resistant to phosphine. Nevertheless, these rapid diagnostics are important in the case of routine treatment in given facilities and commodities that are often fumigated.

ABIOTIC CONDITIONS ARE IMPORTANT Monitoring is essential in phosphine use. When it comes to monitoring, monitoring of the gas concentration is not the only thing that has to be monitored. Other parameters, such as temperature etc. should be monitored in parallel, as phosphine concentration has interesting diurnal circles according to the key abiotic conditions prevailing. Distribution of the gas is also important- even more important is monitoring of distribution. The traditional monitoring techniques are able to provide measurements for specific time intervals and at specific locations within the fumigated facilities. However, the measurements refer to the average concentrations, which are not always representative; in this sense it is the extreme values that are responsible for resistance development and not the average. Extreme dosing and absence of monitoring are key elements that are directly related with resistance development.

REFERENCES Agrafioti P., Athanassiou C. G., Nayak M. K. (2019). Detection of phosphine resistance in stored-product insects in Greece and evaluation of a field resistance test kit. J Stored Prod Res 82: 40-47. Araujo RA, Guedes RNC, Oliveira MGA et al (2008) Enhanced proteolytic and cellulolytic activity in insecticide-resistance strains of the maize weevil, Sitophilus zeamais. J Stored Prod Res 44:354–359 Benhalima, H., Chaudhry, M.Q., Mills, K.A., Price, N.R., 2004. Phosphine resistance in stored-product insects collected from various grain storage facilities in Morocco. J. Stored Prod. Res.40, 241-249. Chandhry M. Q., 2000.Phosphine resistance: a growing threat to an ideal fumigant. Pesticide Outlook, pp: 88-91. Chen, Z., Schlipalius, D., Opit, G., Subramanyam, B., Phillips, T.W., 2015. Diagnostic Molecular Markers for Phosphine Resistance in U.S. populations of Tribolium castaneum and Rhyzopertha dominica. PloS One 10, 0121343. Collins, P.J., Daglish, G.J., Pavic, H., and Kopittke, R.A., 2005. Response of mixed-age cultures of phosphine-resistant and susceptible strains of lesser grain borer, Rhyzopertha dominica, to phosphine at a range of concentrations and exposure periods. J. Stored Prod. Res. 41, 373-385. FAO, 1976. Recommended methods for detection and measurement of resistance of agricultural pests to pesticides - tentative method for adults of some major pest species of stored cereals, with methyl-bromide and phosphine. FAO Method No 16. FAO Plant Prot. Bull. 23, 12–25. Holloway, J., Falk, M.J., Emery, R.N., Nayak, M.K., 2016. Resistance to phosphine in Sitophilus oryzae in Australia: A national analysis of trends and frequencies over time and geographical spreads. J. Stored Prod. Res. 69, 129-137. Kaur, R, Schilipalius, DI, Collins, PJ, Swain, AJ & Ebert, PR (2012) Inheritance and relative dominance, expressed as toxicity response and delayed development, of phosphine resistance in immature stages of Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae). J..Stored Prod. Res. 51, 74- 80. Konemann, CE, Hubhachen, Z, Opit, GP, Gautam, S & Bajracharya, NS., 2017.Phosphine resistance in Cryptolestes ferrugineus(Coleoptera:Laemophloeidae) collected from grain storage facilities in Oklahoma, USA. J. Econ. Entomol.110, 1377-1383. Lorini, I., Collins, P.J., Daglish, G.J., Nayak, M.K., and Pavic, H., 2007. Detection and characterisation of strong resistance to phosphine in Brazilian Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae). Pest Manag. Sci. 63, 358-364. Nayak, M.K., Collins, P.J., 2008. Influence of concentration, temperature and humidity on toxicity of phosphine against strongly phosphine-resistant psocid Liposcelis bostrychophila Badonnel (Psocoptera: Liposcelididae). Pest Manag. Sci. 64, 971-976. Nayak, M.K., Holloway, J.C., Emery, R.N., Pavic, H., Bartlet, J. and Collins, P.J., 2013. Strong resistance to phosphine in the rusty grain beetle, Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae): its characterisation, a rapid assay for diagnosis and its distribution in Australia. Pest. Manag. Sci., 69, 48–53. Nayak, MK, Holloway J, Pavic H, Head M, Reid R and Collins PJ, 2010. Developing strategies to manage highly phosphine resistant populations of rusty grain beetles in large bulk storages in Australia, pp.396-401. Proceedings of the 10th International Working Conference on Stored Product Protection, ed. by Carvalho MO, Fields PG, Adler CS, Arthur FH, Athanassiou CG, Campell JF, et al. Julius-Kuhn Arch, 425, 396 – 401.

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