Host Tree Influences on Longicorn Beetle (Coleoptera: Cerambycidae) Attack in Subtropical Corymbia (Myrtales: Myrtaceae)

Phoracantha longicorn beetles are endemic to Australia, and some species have become significant pests of eucalypts worldwide, yet little is known about their host plant interactions and factors influencing tree susceptibility in Australia. Here, we investigate the host relationships of Phoracantha solida (Blackburn, 1894) on four eucalypt taxa (one pure species and three hybrid families), examining feeding site physical characteristics including phloem thickness, density, and moisture content, and host tree factors such as diameter, height, growth, taper, and survival. We also determine the cardinal and vertical (within-tree) and horizontal (between-tree) spatial distribution of borers. Fewer than 10% of P. solida attacks were recorded from the pure species (Corymbia citriodora subsp. variegate (Hook)), and this taxon also showed the highest survival, phloem thickness, relative growth rate, and bark:wood area. For the two most susceptible taxa, borer severity was negatively correlated with moisture content, and positively related to phloem density. Borers were nonrandomly and nonuniformly distributed within trees, and were statistically aggregated in 32% of plots. More attacks were situated on the northern side of the tree than the other aspects, and most larvae fed within the lower 50 cm of the bole, with attack height positively correlated with severity. Trees with borers had more dead neighbors, and more bored neighbors, than trees without borers, while within plots, borer incidence and severity were positively correlated. Because the more susceptible taxa overlapped with less susceptible taxa for several physical tree factors, the role of primary and secondary chemistries in determining host suitability needs to be investigated. Nevertheless, taxon, moisture content, phloem density, tree size, and mortality of neighboring trees appeared the most important physical characteristics influencing host suitability for P. solida at this site.

Dietary media for mass rearing of rusty grain beetle, Cryptolestes ferrugineus (Stephens) and flat grain beetle, Cryptolestes pusillus (Schonherr) (Coleoptera: Cucujidae)

We developed a suitable diet for mass rearing of Cryptolestes ferrugineus (Stephens) populations under laboratory conditions. Recently, this pest has developed strong level of resistance to phosphine in Australia, and therefore, a significant amount of research has been directed towards its management. In total, nineteen grain-based diets, containing rolled oats, various combinations of cracked grains and flours of wheat, sorghum, maize and barley were tested. Each diet contained a small proportion of wheat germ (4.5% w/w) and torula yeast (0.5% w/w). Experiments were conducted at fixed temperature and relative humidity regimes of 30 ± 2 °C and 70 ± 2%, respectively, and replicated three times. Adults (n = 40) of a laboratory strain of C. ferrugineus were introduced into each diet, removed after 14 days and total numbers of live adult progeny were recorded. The following diets resulted in highest live progeny production: barley flour (95%) (607.67 ± 11.21) = rolled oats (75%) + cracked sorghum (20%) (597.33 ± 33.79) ≥ wheat flour (47.5%) + barley flour (47.5%) (496.67 ± 52.93) > cracked sorghum (95%) (384.00 ± 60.66). The performance of these four diets was then tested with field-collected populations of C. ferrugineus and Cryptolestes pusillus (Schonherr). The diets based on rolled oats + cracked sorghum, wheat flour + barley flour, and barley flour alone consistently produced highest progeny numbers in field-collected populations of both species, with mean progeny numbers ranging from 359.9 to 478.5. The multiplication of C. pusillus was significantly higher than C. ferrugineus on all four diets. Our findings will help in mass rearing of healthy cultures of C. ferrugineus and C. pusillus that will greatly facilitate laboratory and field research and in particular, in developing management tactics for these species.

Developing effective fumigation protocols to manage strongly phosphine-resistant Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae)

BACKGROUND The emergence of high levels of resistance in Cryptolestes ferrugineus (Stephens) in recent years threatens the sustainability of phosphine, a key fumigant used worldwide to disinfest stored grain. We aimed at developing robust fumigation protocols that could be used in a range of practical situations to control this resistant pest. RESULTS Values of the lethal time to kill 99.9% (LT99.9, in days) of mixed-age populations, containing all life stages, of a susceptible and a strongly resistant C. ferrugineus population were established at three phosphine concentrations (1.0, 1.5 and 2.0 mg L−1) and three temperatures (25, 30 and 35 °C). Multiple linear regression analysis revealed that phosphine concentration and temperature both contributed significantly to the LT99.9 of a population (P < 0.003, R2 = 0.92), with concentration being the dominant variable, accounting for 75.9% of the variation. Across all concentrations, LT99.9 of the strongly resistant C. ferrugineus population was longest at the lowest temperature and shortest at the highest temperature. For example, 1.0 mg L−1 of phosphine is required for 20, 15 and 15 days, 1.5 mg L−1 for 12, 11 and 9 days and 2.0 mg L−1 for 10, 7 and 6 days at 25, 30 and 35 °C, respectively, to achieve 99.9% mortality of the strongly resistant C. ferrugineus population. We also observed that phosphine concentration is inversely proportional to fumigation period in regard to the population extinction of this pest. CONCLUSION The fumigation protocols developed in this study will be used in recommending changes to the currently registered rates of phosphine in Australia towards management of strongly resistant C. ferrugineus populations, and can be repeated in any country where this type of resistance appears.

Genetic characterization of field-evolved resistance to phosphine in the rusty grain beetle, Cryptolestes ferrugineus (Laemophloeidae: Coleoptera)

Inheritance of resistance to phosphine fumigant was investigated in three field-collected strains of rusty grain beetle, Cryptolestes ferrugineus, Susceptible (S-strain), Weakly Resistant (Weak-R) and Strongly Resistant (Strong-R). The strains were purified for susceptibility, weak resistance and strong resistance to phosphine, respectively, to ensure homozygosity of resistance genotype. Crosses were established between S-strain × Weak-R, S-strain × Strong-R and Weak-R × Strong-R, and the dose mortality responses to phosphine of these strains and their F1, F2 and F1-backcross progeny were obtained. The fumigations were undertaken at 25 °C and 55% RH for 72 h. Weak-R and Strong-R showed resistance factors of 6.3 × and 505 × compared with S-strain at the LC50. Both weak and strong resistances were expressed as incompletely recessive with degrees of dominance of − 0.48 and − 0.43 at the LC50, respectively. Responses of F2 and F1-backcross progeny indicated the existence of one major gene in Weak-R, and at least two major genes in Strong-R, one of which was allelic with the major factor in Weak-R. Phenotypic variance analyses also estimated that the number of independently segregating genes conferring weak resistance was 1 (nE = 0.89) whereas there were two genes controlling strong resistance (nE = 1.2). The second gene, unique to Strong-R, interacted synergistically with the first gene to confer a very high level of resistance (~ 80 ×). Neither of the two major resistance genes was sex linked. Despite the similarity of the genetics of resistance to that previously observed in other pest species, a significant proportion (~ 15 to 30%) of F1 individuals survived at phosphine concentrations higher than predicted. Thus it is likely that additional dominant heritable factors, present in some individuals in the population, also influenced the resistance phenotype. Our results will help in understanding the process of selection for phosphine resistance in the field which will inform resistance management strategies. In addition, this information will provide a basis for the identification of the resistance genes.

Genetic characterization of field-evolved resistance to phosphine in the rusty grain beetle, Cryptolestes ferrugineus (Laemophloeidae: Coleoptera)

Inheritance of resistance to phosphine fumigant was investigated in three field-collected strains of rusty grain beetle, Cryptolestes ferrugineus, Susceptible (S-strain), Weakly Resistant (Weak-R) and Strongly Resistant (Strong-R). The strains were purified for susceptibility, weak resistance and strong resistance to phosphine, respectively, to ensure homozygosity of resistance genotype. Crosses were established between S-strain × Weak-R, S-strain × Strong-R and Weak-R × Strong-R, and the dose mortality responses to phosphine of these strains and their F1, F2 and F1-backcross progeny were obtained. The fumigations were undertaken at 25 °C and 55% RH for 72 h. Weak-R and Strong-R showed resistance factors of 6.3 × and 505 × compared with S-strain at the LC50. Both weak and strong resistances were expressed as incompletely recessive with degrees of dominance of − 0.48 and − 0.43 at the LC50, respectively. Responses of F2 and F1-backcross progeny indicated the existence of one major gene in Weak-R, and at least two major genes in Strong-R, one of which was allelic with the major factor in Weak-R. Phenotypic variance analyses also estimated that the number of independently segregating genes conferring weak resistance was 1 (nE = 0.89) whereas there were two genes controlling strong resistance (nE = 1.2). The second gene, unique to Strong-R, interacted synergistically with the first gene to confer a very high level of resistance (~ 80 ×). Neither of the two major resistance genes was sex linked. Despite the similarity of the genetics of resistance to that previously observed in other pest species, a significant proportion (~ 15 to 30%) of F1 individuals survived at phosphine concentrations higher than predicted. Thus it is likely that additional dominant heritable factors, present in some individuals in the population, also influenced the resistance phenotype. Our results will help in understanding the process of selection for phosphine resistance in the field which will inform resistance management strategies. In addition, this information will provide a basis for the identification of the resistance genes.

Developing effective fumigation protocols to manage strongly phosphine-resistant Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae)

BACKGROUND The emergence of high levels of resistance in Cryptolestes ferrugineus (Stephens) in recent years threatens the sustainability of phosphine, a key fumigant used worldwide to disinfest stored grain. We aimed at developing robust fumigation protocols that could be used in a range of practical situations to control this resistant pest. RESULTS Values of the lethal time to kill 99.9% (LT99.9, in days) of mixed-age populations, containing all life stages, of a susceptible and a strongly resistant C. ferrugineus population were established at three phosphine concentrations (1.0, 1.5 and 2.0 mg L−1) and three temperatures (25, 30 and 35 °C). Multiple linear regression analysis revealed that phosphine concentration and temperature both contributed significantly to the LT99.9 of a population (P < 0.003, R2 = 0.92), with concentration being the dominant variable, accounting for 75.9% of the variation. Across all concentrations, LT99.9 of the strongly resistant C. ferrugineus population was longest at the lowest temperature and shortest at the highest temperature. For example, 1.0 mg L−1 of phosphine is required for 20, 15 and 15 days, 1.5 mg L−1 for 12, 11 and 9 days and 2.0 mg L−1 for 10, 7 and 6 days at 25, 30 and 35 °C, respectively, to achieve 99.9% mortality of the strongly resistant C. ferrugineus population. We also observed that phosphine concentration is inversely proportional to fumigation period in regard to the population extinction of this pest. CONCLUSION The fumigation protocols developed in this study will be used in recommending changes to the currently registered rates of phosphine in Australia towards management of strongly resistant C. ferrugineus populations, and can be repeated in any country where this type of resistance appears. © 2014 Commonwealth of Australia. Pest Management Science © 2014 Society of Chemical Industry

Susceptibility to sulfuryl fluoride and lack of cross-resistance to phosphine in developmental stages of the red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae)

BACKGROUND Our aim was to ascertain the potential of sulfuryl fluoride (SF) as an alternative fumigant to manage phosphine-resistant pests. We tested the susceptibility of all life stages of red flour beetle, Tribolium castaneum (Herbst), to SF and assessed the presence of cross-resistance to this fumigant in phosphine-resistant strains of this species. RESULTS Analysis of dose–response data indicated that the egg was the stage most tolerant to SF under a 48 h exposure period. At LC50, eggs were 29 times more tolerant than other immature stages and adults, and required a relatively high concentration of 48.2 mg L−1 for complete mortality. No significant differences in tolerance to SF were observed among the three larval instars, pupae and adults, and all of these stages were controlled at a low concentration of 1.32 mg L−1. Phosphine-resistant strains did not show cross-resistance to SF. CONCLUSION Our research concluded that the current maximum registered rate of SF, 1500 gh m−3, is adequate to control all the post-embryonic life stages of T. castaneum over a 48 h fumigation period, but it will fail to achieve complete mortality of eggs, indicating the risk of some survival of eggs under this short exposure period. As there is no cross-resistance to SF in phosphine-resistant insects, it will play a key role in managing phosphine resistance in stored-grain insect pests. © 2014 Commonwealth of Australia. Pest Management Science © 2014 Society of Chemical Industry

New synonymies for Australian Cleridae (Coleoptera)

The following synonymies are proposed based on examination of primary types (lectotypes are designated for all taxa except those marked with a '*'): Lemidia spinnipennis Lea, 1907 syn. n. and Lemidia bicolor Schenkling, 1906 syn. n. = Lemidia biaculeata (Westwood); Lemidia mastersi Lea, 1907 syn. n. = Lemidia circumcincta Schenkling, 1906; Lemidia albonotata Pic, 1941 syn. n. = Lemidia laticeps Lea, 1907; Lemidia australiae Lea, 1907 syn. n. = Lemidia maculata Schenkling, 1902; Lemidia bilineatra Lea, 1907 syn. n. = Lemidia maculicollis Gorham, 1877; Lemidia decolor Pic, 1941 syn. n. = Lemidia munda Blackburn, 1892; *Phlogistus conspiciendus Elston, 1926 syn. n. = Mimolesterus ventralis (Westwood); Thanasimus cursorius Westwood, 1853 syn. n. and Stigmatium dispar Kuwert, 1894 syn. n. = Stigmatium acerbum (Newman); Stigmatium fasciatoventre Chevrolat, 1874 syn. n., Stigmatium flavescens Chevrolat, 1874 syn. n. and *Xestonotus eximius Kuwert, 1894 syn. n. = Stigmatium laevium Macleay, 1872; Stigmatium versipelle Gorham, 1876 syn. n. and Xestonotus (Cyclotomocerus) australicus Kuwert, 1894 syn. n. = Stigmatium varipes Chevrolat, 1876; Tarsostenus pulcher Macleay, 1872 syn. n. = *Tarsostenus carus (Newman, 1840). The available name Tarsosternus pulcher Macleay, 1872 is deemed a lapsus calami and emended to Tarsostenus pulcher Macleay, 1872.

Developmental biology and prey preference of Diomus notescens Blackburn (Coleoptera: Coccinellidae): A predator of Aphis gossypii Glover (Hemiptera: Aphididae)

The minute two-spotted ladybeetle, Diomus notescens Blackburn is a common predator of aphids and other pests in Australian agricultural crops, however little is known about the biology of D. notescens. The aim of this study was to provide information on the life cycle of this predator and improve our understanding of its biological control potential, particularly against one of the major pests of cotton, Aphis gossypii Glover. In laboratory experiments, juvenile development, prey consumption, as well as adult lifespan and fecundity were studied. Results from this study revealed that D. notescens could successfully complete development on A. gossypii, which at 25 °C required 21 days and during this period they each consume 129 ± 5.2 aphids. At 25 °C adult lifespan was 77 ± 9.6 days, with a mean daily prey consumption of 28 ± 1.8 aphids and a mean daily fecundity of 8 ± 0.5 eggs. Net reproductive rate was estimated as 187 ± 25.1 females and the intrinsic rate of increase was estimated as 0.14. Juvenile development was recorded at four constant temperatures (15, 21, 26 and 27 °C) and using a linear model, the lower threshold for D. notescens development was estimated to be 10 ± 0.6 °C with 285 ± 4.7 degree days required to complete development. A prey choice experiment studying predation rates revealed a strong preference for A. gossypii nymphs compared to Bemisia tabaci Gennadius eggs.

Lesser auger beetle Heterobostrychus aequalis(Coleoptera: Bostrichidae) in Australia: absent or elusive?

The lesser auger beetle, Heterobostrychus aequalis (Waterhouse), is a serious pest of seasoned hardwood timber throughout the Oriental region and several areas beyond. Some early collection records of H. aequalis from Australia in the 1950s and 1960s indicated that the insect was present in northern Queensland, but no confirmed breeding population has been found in the past few decades suggesting either that it may have not established permanently or it is difficult to detect. The ambiguity about the breeding status of the pest in Australia has caused confusion for regulating authorities needing to respond to each new post-border detection. We examined records of H. aequalis in Australian insect collections and from intensive plant pest surveillance activities in Queensland and northern Australia over the past 48 years to resolve this confusion. Until very recently, available evidence suggested that H. aequalis was not established in Australia, despite multiple introductions and apparently suitable climate and hosts. Collection records of the pest are predominantly linked to intercepted items or are recorded as of unknown origin, and no established populations have been found during many years of targeted surveillance. However, a detection of H. aequalis in suburban Cairns, north Queensland, in late 2013 and two more in mid-2015 in the same general locality do not appear to be linked to any imported material, indicating that there is at least a tenuously established local population. Investigations are underway to confirm this, but the insect is not widely established in Australia and, if present, remains elusive. Our recommended response to any future detection of H. aequalis is to fumigate or destroy the infested material, conduct tracing enquiries and limit surveys to the immediate vicinity of the detection.

Influence of phosphine resistance genes on flight propensity and resource location in Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae): the landscape for selection

Phosphine resistance in Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) has evolved through changes to enzymes involved in basic metabolic pathways. These changes impose metabolic stress and could affect energy-demanding behaviours. We therefore tested whether phosphine resistance alleles impact the movement of these insects in their quest for new resources. We measured walking and flight parameters of four T. castaneum genotypes: (1) a field-derived population, (2) a laboratory cultured, phosphine-susceptible reference strain, (3) a laboratory cultured, phosphine-resistant reference strain, and (4) a resistant introgressed strain that is almost identical genetically to the susceptible population. The temporal pattern of flight was identical across all populations, but resistant beetles took flight significantly less, walked more slowly, and located resources less successfully than did susceptible beetles. Also, the field-derived beetles (proved not to be carrying resistance genes) walked significantly faster and more directly towards food resources, and had a higher propensity for flight when compared to the susceptible laboratory beetles. These negative effects suggest survival of beetles with the resistance alleles will be compromised should they leave phosphine application sites. The field for selection therefore extends beyond the site at which phosphine fumigant imposed its effect, and other mutations are also likely to be affected in this way.

Determining phosphine resistance in rust red flour beetle, Tribolium castaneum (Herbst.) (Coleoptera : Tenebrionidae) populations from Turkey

Fumigation with phosphine gas is the primary method of controlling stored grain pests. In Turkey, phosphine has been used extensively since the 1950's. Even though high levels of phosphine resistance have been detected in several key stored products pests across the world, it has never been studied in Turkey despite this long history of phosphine use. High-level phosphine resistance has been detected and genetically characterised previously in the rust red flour beetle, Tribolium castaneum in other countries. Since this pest is also a common problem in stored grain environment in Turkey, the current study was undertaken for the first time, to investigate the distribution and strength of phosphine resistance in T. castaneum. Four strains of T. castaneum were tested through bioassays for determining the weak and strong phosphine resistance phenotypes on the basis of the response of adults to discriminating phosphine concentrations of 0.03 mg/L and 0.25 mg/L, for 20 hour exposures respectively. Phenotype testing showed all strains exhibited some level of phosphine resistance with a maximum level of 196 fold. Sequencing and genetic testing of seven field-collected strains showed that all of them carried a strong resistance allele in at the rph2 locus similar to the one previously reported. Our results show that strong resistance to phosphine is common in Turkish strains of T. castaneum.

Random Mating Between Two Widely Divergent Mitochondrial Lineages of Cryptolestes ferrugineus (Coleoptera: Laemophloeidae): A Test of Species Limits in a Phosphine-Resistant Stored Product Pest

Effective pest management relies on accurate delimitation of species and, beyond this, on accurate species identification. Mitochondrial COI sequences are useful for providing initial indications in delimiting species but, despite acknowledged limitations in the method, many studies involving COI sequences and species problems remain unresolved. Here we illustrate how such impasses can be resolved with microsatellite and nuclear sequence data, to assess more directly the amount of gene flow between divergent lineages. We use a population genetics approach to test for random mating between two 8 ± 2% divergent COI lineages of the rusty grain beetle, Cryptolestes ferrugineus (Stephens). This species has become strongly resistant to phosphine, a fumigant used worldwide for disinfesting grain. The possibility of cryptic species would have significant consequences for resistance management, especially if resistance was confined to one mitochondrial lineage. We find no evidence of restricted gene flow or nonrandom mating across the two COI lineages of these beetles, rather we hypothesize that historic population structure associated with early Pleistocene climate changes likely contributed to divergent lineages within this species.

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.

Scat contents of the spotted-tailed quoll Dasyurus maculatus in the New England gorges, north-eastern New South Wales

In 1313 scats of the spotted-tailed quoll Dasyurus maculatus, collected over 5 years from the gorge country of north-eastern New South Wales, the most frequent and abundant items were derived from mammals and a restricted set of insect orders. These quolls also ate river-associated items: waterbirds, eels, crayfish, aquatic molluscs and even frogs. Macropods contributed most of the mammal items, with possums, gliders and rodents also being common. Some food, particularly from macropods and lagomorphs, had been scavenged (as shown by fly larvae). The most frequent invertebrates were three orders of generally large insects Coleoptera, Hemiptera and Orthoptera, which were most frequent in summer and almost absent in winter scats. Monthly mean numbers of rodent and small dasyurid items per scat were inversely related to these large insects in scats. The numbers of reptile items were inversely related to the numbers of mammal (especially arboreal and small terrestrial mammal) items per scat, thus types of items interacted in their occurrences in monthly scat samples. Frequencies of most vertebrate items showed no seasonal, but much year-to-year, variation. This quoll population ate four main types of items, each requiring different skills to obtain: they hunted arboreal marsupials (possibly up trees), terrestrial small mammals and reptiles (on the ground), and seasonally available large insects (on trees or the ground), and scavenged carcases, mostly of large mammals but also birds and fishes (wherever they could find them).