Ecological Networks in Stored Grain: Key Postharvest Nodes for Emerging Pests, Pathogens, and Mycotoxins

Wheat is at peak quality soon after harvest. Subsequently, diverse biota use wheat as a resource in storage, including insects and mycotoxin-producing fungi. Transportation networks for stored grain are crucial to food security and provide a model system for an analysis of the population structure, evolution, and dispersal of biota in networks. We evaluated the structure of rail networks for grain transport in the United States and Eastern Australia to identify the shortest paths for the anthropogenic dispersal of pests and mycotoxins, as well as the major sources, sinks, and bridges for movement. We found important differences in the risk profile in these two countries and identified priority control points for sampling, detection, and management. An understanding of these key locations and roles within the network is a new type of basic research result in postharvest science and will provide insights for the integrated pest management of high-risk subpopulations, such as pesticide-resistant insect pests.

Managing resistance to chemical treatments in stored products pests

This review focuses on key trends in resistance to chemical treatments in stored product pests, and advances in resistance management, with an emphasis on resistance to the fumigant phosphine. Findings: Phosphine resistance continues to be a major concern. In particular, phosphine resistance in Cryptolestes ferrugineus has emerged as a serious issue, with some populations exhibiting the strongest level detected so far for this fumigant. In response, a 'quick knock down test' has been established to deliver industry and scientists 'same day' advice on the resistance status of field samples; sulfuryl fluoride is being developed as a 'resistance breaker' and phosphine dosages are being revised to manage this problem. There has been major progress in identifying the genes responsible for phosphine resistance and the development of molecular resistance diagnostics for key pests. Several studies on Rhyzopertha dominica have demonstrated that molecular screening can be used to determine the frequency of resistance alleles in samples collected from farm storages. Despite on-going research in several pests, there is no definitive answer to the question of whether there is a fitness cost associated phosphine resistance, with some studies showing a clear cost and others none. Evidence continues to emerge of resistance to grain protectants, including the juvenile hormone analogue methoprene. The development and adoption of spinosad, as a next generation 'green' treatment, and the use of protectant combinations provides opportunities to counter the problem of protectant resistance.Directions for future research: A uniform set of protocols should be developed for phosphine resistance detection for all major species. It should combine 'quick tests' and molecular diagnostics to be adopted internationally. Research is required on the establishment of a decision making system that integrates newly developed grain protectants and fumigants, other alternative control methods, as well as an accurate and rapid resistance detection system for early warning of the emergence of new resistances.

The role of spiders as predators of two lepidopteran Brassica pests

Spiders are thought to play a significant role in limiting pest outbreaks in agroecosystems such as vineyards, orchards and cotton. The diversity and impact of spiders in vegetable crops are less well understood, although there is evidence that predators may be important for suppression of lepidopteran pests in Brassica crops, particularly early in the season before parasitoids become established. Sampling was conducted in early season plantings of Brassicas in the Lockyer Valley (South East Queensland, Australia) in order to determine the most commonly occurring spider families. The most numerous were Theridiidae, which were more strongly associated with cauliflower and poorly associated with cabbage. The Lycosidae and Clubionidae/Miturgidae (formerly in the ‘catch-all’ family Clubionidae) also occurred commonly. Lycosidae (and to a lesser extent Salticidae) had above average abundance in Chinese cabbage and below average abundance in broccoli compared with average abundance for these spider families; Clubionidae/Miturgidae had above average abundance in cauliflower. Laboratory studies were then conducted to explore the predatory capacity of these three most commonly occurring spider families. All three were capable of feeding on larvae of the diamondback moth, Plutella xylostella (Linnaeus), and cabbage cluster caterpillar, Crocidolomia pavonana (Fabricius), under laboratory conditions. Theridiidae, which are thought to prey on small pests such as leafhoppers and aphids, were able to successfully attack larvae up to five times their body size. Predation rates varied from an average of 1.7 (SE = 0.47) (1.6 control corrected) larvae consumed over a 24 h period in the case of the Theridiidae, to 3.3 (SE = 0.60) larvae for the Clubionidae/Miturgidae.

The grand challenge of food security - general lessons from a comprehensive approach to protecting stored grain from insect pests in Australia and India

We outline a philosophical approach to Grand Challenge projects, with particular reference to our experience in our food security project involving the protection of stored grain from insect attack in two countries on different continents. A key consideration throughout has been the management of resistance in these pests to the valuable fumigant phosphine. Emphasis is given to the chain of research issues that required solution and the assembly of a well-integrated team, overlapping in skills for effective communication, in each country to solve the problems identified along that chain. A crucial aspect to maintaining direction is the inclusion of key end users in all deliberations, as well as the establishment and maintenance of effective outlets for the dissemination of practical recommendations. We finish with a summary of our achievements with respect to our approach to this food security Grand Challenge.

Status of resistance to phosphine in insect pests of stored products in Vietnam

In response to numerous reports of failures to control insect pests of stored products with phosphine in Vietnam, a national survey for resistance to this key fumigant was undertaken in 2009–2011. Data from a more limited survey undertaken by the authors in 2002 in northern Vietnam are also presented. Samples collected in the 2002 survey (Sitophilus oryzae, n=8; Tribolium castaneum, n=8) were tested using a full dose- response assay, while for the 2009–11 survey, F1 generations were tested for resistance with two discriminating dosages of phosphine to detect frequency of weak and strong resistance phenotypes. Compared with a susceptible reference strain, in 2002, resistance to phosphine was indicated in six T. castaneum samples but only two of S. oryzae. Resistance factor, however, did not exceed 2.8-fold in T. castaneum and 1.7 in S. oryzae indicating relatively low frequency and weak expression of resistance. In 2009–11 survey, 176 samples were collected from a range of food and feed storages along the supply chain and from all major regions of Vietnam (125 sites). Rhyzopertha dominica and S. oryzae were the most common species found infesting stored commodities. Resistance was detected at high frequency in all the species. Weak and strong resistance phenotype frequencies were, respectively: Cryptolestes ferrugineus (37 and 58%, n=19), R. dominica (1.5 and 97%, n=65), S. oryzae (34 and 59%, n=82) and T. castaneum (70 and 30%, n=10). Strong resistance phenotype was detected in all the major regions and all parts of the supply chain but frequency was the highest in central storages and animal feed establishments. The increase in frequency and strength of resistance to phosphine in the eight years between the two surveys has been rapid and dramatic. The survey demonstrates the threat of resistance to grain protection in Vietnam and highlights the need for training of fumigators, and the development and adoption of phosphine resistance management tactics nationally.

The importance of cotton disease surveys in Queensland for monitoring endemic diseases and detecting new pathogens and pests

The cotton industry in Australia funds biannual disease surveys conducted by plant pathologists. The objective of these surveys is to monitor the distribution and importance of key endemic pests and record the presence or absence of new or exotic diseases. Surveys have been conducted in Queensland since 2002/03, with surveillance undertaken by experienced plant pathologists. Monitoring of endemic diseases indicates the impact of farming practices on disease incidence and severity. The information collected gives direction to cotton disease research. Routine diagnostics has provided early detection of new disease problems which include 1) the identification of Nematospora coryli, a pathogenic yeast associated with seed and internal boll rot; and 2) Rotylenchulus reniformis, a plant-parasitic nematode. This finding established the need for an intensive survey of the Theodore district revealing that reniform was prevalent across the district at populations causing up to 30% yield loss. Surveys have identified an exotic defoliating strain (VCG 1A) and non-defoliating strains of Verticillium dahliae, which cause Verticillium wilt. An intensive study of the diversity of V. dahliae and the impact these strains have on cotton are underway. Results demonstrate the necessity of general multi-pest surveillance systems in broad acre agriculture in providing (1) an ongoing evaluation of current integrated disease management practices and (2) early detection for a suite of exotic pests and previously unknown pests.

Induced Resistance: Potential for Control of Postharvest Diseases of Horticultural Crops

Fortunately, plants have developed highly effective mechanisms with which to defend themselves when attacked by potentially disease-causing microorganisms. If not, then they would succumb to the many pathogenic fungi, bacteria, viruses, nematodes and insect pests, and disease would prevail. These natural defence systems of plants can be deliberately activated to provide some protection against the major pathogens responsible for causing severe yield losses in agricultural and horticultural crops. This is the basis of what is known as ‘induced’ or ‘acquired’ disease resistance in plants. Although the phenomenon of induced resistance has been known amongst plant pathologists for over 100 years, its inclusion into pest and disease management programmes has been a relatively recent development, ie. within the last 5 years. This review will discuss very briefly some of the characteristics of the induced resistance phenomenon, outline some of the advantages and limitations to its implementation and provide some examples within a postharvest pathology context. Finally some approaches being investigated by the fruit pathology team at DPI Indooroopilly and collaborators will be outlined.

Effect of fumigation temperature on the efficacy of phosphine against strongly resistant psocids liposcelis bostrychophila (Pscodoptera: Liposcelididae)

Species of Liposcelis psocids have emerged as major pests of stored grain in Australia in recent years. Several populations have been detected with high resistance to phosphine, the major chemical treatment. Highest resistance has been detected in the cosmopolitan species Liposcelis bostrychophila. As part of a national resistance management strategy to maintain the viability of phosphine, we are developing minimum effective dosage regimes (concentration x time) required to control all life stages of resistant L. bostrychophila at a range of grain temperatures. Four concentrations of phosphine, 0.1, 0.17, 0.3 aid 1 mg/L, were evaluated for their effectiveness against strongly resistant L. bostrychophila at a series of fumigation temperatures: 20, 25, 30 and 35°C. Results were recorded as the least number of days taken to achieve population extinction. We found that, at any fixed concentration of phosphine, time to population extinction decreased as fumigation temperature increased from 20 to 30°C. For example, at 0.1 mg/L, it took more than 14 days at 20°C to completely control these insects, whereas at 30°C it took only seven days. Increase in fumigation temperature from 25OC to 30°C dramatically reduced the exposure period needed to achieve population extinction of resistant psocids. For example, a dose of 0.17 mg/L over six days at 30°C completely controlled strongly resistant L. bostrychophila populations that can survive at 1 mg/L and 25°C over the same exposure period. Findings from our study will be used to formulate recommendations for registered dosage rates and fumigation periods for use in Australia.

Cane Weevil Borer, Rhabdoscelus obscurus (Coleoptera: Curculionidae), a pest of palms in Northern Queensland, Australia

In recent years the cultivation of ornamental palms (Arecaceae) has increased markedly in northern Queensland. Consequently, several insects have become important pests, particularly Rhabdoscelus obscurus (Boisduval), the cane weevil borer. The larvae of this beetle feed on various species of palms, making the plants unsaleable. Death or lodging of the trees may also result. This paper documents its pest status, derived from information in the literature and from consultation with local growers.

A sampling technique for two-spotted mite in papaya

Two-spotted mite, Tetranychus urticae Koch, was until recently regarded as a minor and infrequent pest of papaya in Queensland through the dry late winter/early summer months. The situation has changed over the past 4-5 years, so that now some growers consider spider mites significant pests all year round. This altered pest status corresponded with a substantial increase in the use of fungicides to control black spot (Asperisporium caricae). A project was initiated in 1998 to examine the potential reasons for escalating mite problems in commercially-grown papaya, which included regular sampling over a 2 year period for mites, mite damage and beneficial arthropods on a number of farms on the wet tropical coast and drier Atherton Tableland. Differences in soil type, papaya variety, chemical use and some agronomic practices were included in this assessment. Monthly visits were made to each site where 20 randomly-selected plants from each of 2 papaya lines (yellow and red types) were surveyed. Three leaves were selected from each plant, one from each of the bottom, middle and top strata of leaves. The numbers of mobile predators were recorded, along with visual estimates of the percentage and age of mite damage on each leaf. Leaves were then sprayed with hairspray to fix the mites and immature predators to the leaf surface. Four leaf disks, 25 mm in diameter, were then punched from each leaf into a 50 ml storage container with a purpose-built disk-cutting tool. Disks from each leaf position were separated by tissue paper, within the container. On return to the laboratory, each leaf disk was scrutinised under a binocular microscope to determine the numbers of two-spotted mites and eggs, predatory mites and eggs, and the immature stages of predatory insects (mainly Stethorus, Halmus and lacewings). A total of 2160 leaf disks have been examined each month. All data have been entered into an Access database to facilitate comparisons between sites.

Pheromone-mediated mass trapping and population diversion as strategies for suppressing Carpophilus spp. (Coleoptera: Nitidulidae) in Australian stone fruit orchards

1 Five experiments were conducted during 1995-99 in stone fruit orchards on the Central Coast and in inland New South Wales, Australia, on the use of synthetic aggregation pheromones and a coattractant to suppress populations of the ripening fruit pests Carpophilus spp. (Coleoptera: Nitidulidae). 2 Perimeter-based suppression traps baited with pheromone and coattractant placed at 3m intervals around small fruit blocks, caught large numbers of Carpophilus spp. Very small populations of Carpophilus spp. occurred within blocks, and fruit damage was minimal. 3 Carpophilus spp. populations in stone fruit blocks 15-370m from suppression traps were also small and non-damaging, indicating a large zone of pheromone attractivity. 4 Pheromone/coattractant-baited suppression traps appeared to divert Carpophilus spp. from nearby (130 m) ripening stone fruit. Ten metal drums containing decomposing fruit, baited with pheromone and treated with insecticide, attracted Carpophilus spp. and appeared to reduce populations and damage to ripening fruit at distances of 200-500 m. Populations and damage were significantly greater within 200m of the drums and may have been caused by ineffective poisoning or poor quality/overcrowding of fruit resources in the drums. 5 Suppression of Carpophilus spp. populations using synthetic aggregation pheromones and a coattractant appears to be a realistic management option in stone fruit orchards. Pheromone-mediated diversion of beetle populations from ripening fruit may be more practical than perimeter trapping, but more research is needed on the effective range of Carpophilus pheromones and the relative merits of trapping compared to attraction to insecticide-treated areas.

Volatile compounds from the flowers of spathiphyllum Cannaefolium

Headspace analysis and solvent extraction of the pollenbearing flower spike of Spathiphyllum cannaefolium have been conducted by GC-MS, to determine the basis of the flower spike’s attractancy to certain fruit-fly species. The major components were benzyl acetate, methyleugenol, methylchavicol, p-methoxybenzyl acetate and fatty acids. Benzyl acetate is known to be attractive to D. cueurbitae, D. dorsalis and C. capitata (representing the three different ‘male-lure categories’) and methyleugenol (one of these male-lures) attracts D. cacuminatus, D. dorsalis and D. occipitalis. Thus the odoriferous flowerspike exhibits wide ranging attractancy and hence Spathiphyllum cannaefolium may have some application as a fruit-fly control measure for small orchards where ‘methyleugenol-attracted’ species (e.g. D. cacuminatus, D. dorsalis, D. occipitalis) are the dominant pests.

Phenology of Carpophilus spp. (Coleoptera: Nitidulidae)in stone fruit orchards as determined by pheromone trapping: Implications for prediction of crop damage

Traps baited with synthetic aggregation pheromone and fermenting bread dough were used to monitor seasonal incidence and abundance of the ripening fruit pests, Carpophilus hemipterus (L.), C. mutilatus Erichson and C. davidsoni Dobson in stone fruit orchards in the Leeton district of southern New South Wales during five seasons (1991-96). Adult beetles were trapped from September-May, but abundance varied considerably between years with the amount of rainfall in December-January having a major influence on population size and damage potential during the canning peach harvest (late February-March). Below average rainfall in December-January was associated with mean trap catches of < 10 beetles/trap/week in low dose pheromone traps during the harvest period in 1991/92 and 1993/94 and no reported damage to ripening fruit. Rainfall in December-January 1992/93 was more than double the average and mean trap catches ranged from 8-27 beetles/week during the harvest period with substantial damage to the peach crop. December-January rainfall was also above average in 1994/95 and 1995/96 and means of 50-300 beetles/trap/week were recorded in high dose pheromone traps during harvest periods. Carpophilus spp. caused economic damage to peach crops in both seasons. These data indicate that it may be possible to predict the likelihood of Carpophilus beetle damage to ripening stone fruit in inland areas of southern Australia, by routine pheromone-based monitoring of beetle populations and summer temperatures and rainfall.

Genetic variability of Tomato spotted wilt virus in Australia and validation of real time RT-PCR for its detection in single and bulked leaf samples

The potential for large-scale use of a sensitive real time reverse transcription polymerase chain reaction (RT-PCR) assay was evaluated for the detection of Tomato spotted wilt virus (TSWV) in single and bulked leaf samples by comparing its sensitivity with that of DAS-ELISA. Using total RNA extracted with RNeasy® or leaf soak methods, real time RT-PCR detected TSWV in all infected samples collected from 16 horticultural crop species (including flowers, herbs and vegetables), two arable crop species, and four weed species by both assays. In samples in which DAS-ELISA had previously detected TSWV, real time RT-PCR was effective at detecting it in leaf tissues of all 22 plant species tested at a wide range of concentrations. Bulk samples required more robust and extensive extraction methods with real time RT-PCR, but it generally detected one infected sample in 1000 uninfected ones. By contrast, ELISA was less sensitive when used to test bulked samples, once detecting up to 1 infected in 800 samples with pepper but never detecting more than 1 infected in 200 samples in tomato and lettuce. It was also less reliable than real time RT-PCR when used to test samples from parts of the leaf where the virus concentration was low. The genetic variability among Australian isolates of TSWV was small. Direct sequencing of a 587 bp region of the nucleoprotein gene (S RNA) of 29 isolates from diverse crops and geographical locations yielded a maximum of only 4.3% nucleotide sequence difference. Phylogenetic analysis revealed no obvious groupings of isolates according to geographic origin or host species. TSWV isolates, that break TSWV resistance genes in tomato or pepper did not differ significantly in the N gene region studied, indicating that a different region of the virus genome is responsible for this trait.

Minimising mortality in field and pot experiments using Heteronyx piceus Blanchard (Coleoptera: Scarabaeidae) eggs and larvae

The establishment of experimental populations of scarab larvae using eggs and early instar larvae has proven to be difficult for many researchers. Despite this, little work has been published examining ways to optimise establishment under artificial conditions. In this experiment, we examined the effect of shade and irrigation on the establishment of Heteronyx piceus Blanchard larvae introduced into pots as eggs and first-, second- and third-instar larvae to optimise artificial infestation techniques. The most important factor affecting larval establishment was the life stage introduced. Establishment of eggs and first instars was very low, with only 21% of eggs and 11% of first-instar larvae establishing. In contrast, 82% of second-instar larvae and 84% of third-instar larvae established successfully. The addition of shade marginally improved overall survival from 45% in the unshaded pots to 53% in the shaded pots. However, most of this increase was in the eggs and first instars. Irrigation did not improve survival. These results suggest that when introducing scarab larvae to field or pot experiments, second- or thirdinstar larvae should be used to maximise establishment. The provision of shade and supplementary irrigation is optional.