Evidence for beetle pollination in the African grassland sugarbushes (Protea: Proteaceae)

Most lineages in the African genus Protea consist of species with large unscented flowers pollinated principally by birds, and several of these lineages also show evidence of shifts to rodent pollination, associated with concealed yeasty-scented flowerheads. In this study we investigated the hypothesis that brightly coloured and fruity-scented flowerheads of four Protea species (P. caffra, P. simplex, P. dracomontana and P. welwitschii) represent a novel shift from bird to insect pollination in a grassland lineage in the genus. These species are visited by a wide range of insects, but cetoniine beetles were found to be the most important pollinators because of their abundance, size and relatively pure pollen loads. Three of the four putatively insect-pollinated Protea species have flowers presented at ground level, and experiments showed that cetoniine beetles preferred inflorescences at ground level to those artificially elevated to the height of shrubs and small trees. Relative to insects, birds were infrequent visitors to all of the study species. The nectar of all the study species contained xylose, as documented previously in bird- and rodent-pollinated Protea species, suggesting that this is a phylogenetically conserved trait. However, the very low concentration of nectar (ca. 8%), short nectar-stigma distance and the fruity scent of florets appear to be traits that are associated with specialisation for pollination by cetoniine beetles.     

Thermal death kinetics of red flour beetle (Coleoptera: Tenebrionidae)

While developing radio frequency heat treatments for dried fruits and nuts, we used a heating block system developed by Washington State University to identify the most heat-tolerant life stage of red flour beetle, Tribolium castaneum (Herbst), and to determine its thermal death kinetics. Using a heating rate of 15 degrees C/min to approximate the rapid heating of radio frequency treatments, the relative heat tolerance of red flour beetle stages was found to be older larvae > pupae and adults > eggs and younger larvae. Lethal exposure times for temperatures of 48, 50, and 52 degrees C for the most heat-tolerant larval stage were estimated using a 0.5th order kinetic model. Exposures needed for 95% mortality at 48 degrees C were too long to be practical (67 min), but increasing treatment temperatures to 50 and 52 degrees C resulted in more useful exposure times of 8 and 1.3 min, respectively. Red flour beetle was more sensitive to changes in treatment temperature than previously studied moth species, resulting in red flour beetle being the most heat-tolerant species at 48 degrees C, but navel orangeworm, Amyelois transitella (Walker), being most heat tolerant at 50 and 52 degrees C. Consequently, efficacious treatments for navel orangeworm at 50-52 degrees C also would control red flour beetle.     

Insecticidal potential of natural zeolite and diatomaceous earth formulations against rice weevil (Coleoptera: Curculionidae) and red flour beetle (Coleoptera: Tenebrionidae)

Insecticidal potential of natural zeolites and diatomaceous earths originating from Serbia against Sitophilus oryzae (L.) and Tribolium castaneum (Herbst) was evaluated. Two natural zeolite formulations (NZ and NZ Modified) were applied to wheat at rates of 0.50, 0.75, and 1.0 g/kg, while two diatomaceous earth (DE) formulations (DE S-1 and DE S-2) were applied at rates of 0.25, 0.50, 0.75, and 1.0 g/kg. A bioassay was conducted under laboratory conditions: temperature of 24 +/- 1 degrees C, relative humidity in the range 50-55%, in tests with natural zeolites, and 60-65%, in tests with DEs, and in all combinations for progeny production. Mortality was assessed after 7, 14, and 21 d of insect contact with treated wheat, and the total mortality after an additional 7-d recovery on untreated broken wheat. Progeny production was also assessed after 8 wk for S. oryzae and 12 wk for T. castaneum. The highest mortality for S. oryzae and T. castaneum was found after the longest exposure period and 7 d of recovery, on wheat treated with NZ at the highest rate and DEs at rates of 0.50 -1.0 g/kg. Progeny reduction higher than 90% was achieved after 14 and 21 d of contact of both beetle pests with wheat treated with DE S-1 at 0.50-1.0 g/kg and DE S-2 at 0.75-1.0 g/kg, while the same level of reduction was achieved only for T. castaneum after its contact with the highest rate of NZ formulation. NZ Modified, applied even at the highest rate, revealed much lower insecticidal potential.     

Potential resistance of crape myrtle cultivars to flea beetle (Coleoptera: Chrysomelidae) and Japanese beetle (Coleoptera: Scarabaeidae) damage

Field and laboratory studies were conducted to identify potential resistance among crape myrtles, Lagerstroemia spp., to Japanese beetle, Popillia japonica Newman and to flea beetles, Altica spp. Damage ratings revealed variation among cultivars in susceptibility to beetle feeding. Cultivars with Lagerstroemia fauriei Koehne in their parentage exhibited the least amount of damage in choice and no-choice experiments, with few exceptions. The data indicate that both beetle species cause more feeding damage on certain cultivars of Lagerstroemia indica L., such as 'Country Red', 'Twilight', and 'Carolina Beauty' than interspecific cultivars with L. fauriei in their parentage, such as 'Natchez', 'Tonto', and 'Muskogee'. When comparing the effect of parentage on all of the major pests of crape myrtle, L. faurei confers resistance to all pests except crape myrtle aphid. No correlation was found between leaf toughness, leaf color, and leaf nutrients in estimating flea beetle cultivar preference. With this information, growers can more effectively target scouting measures to the most susceptible cultivars. and breeders can select plants that will require the fewest chemical inputs.     

Evidence for limited spatial spread in an exotic longhorn beetle, Tetropium fuscum (Coleoptera: Cerambycidae)

The longhorn beetle Tetropium fuscum F. (Coleoptera: Cerambycidae) has become established in Nova Scotia, Canada, where it coexists with Tetropium cinnamopterum Kirby. The two Tetropium species share a similar ecological niche and use the same volatile cues for mate attraction. Exotic T. fuscum was introduced near Halifax, Nova Scotia, in approximately 1990, but the rate of its spread 20 yr later has not been documented. We report a large-scale, 3-yr study that investigates the distribution of T. fuscum relative to its site of introduction. Traps baited with male-produced pheromone and host volatiles were used to estimate the relative abundance of the two Tetropium species. Adult T. fuscum emerged 1-2 wk earlier than T. cinnamopterum each year between 2008 and 2010. The spatial distribution of T. fuscum was characterized by a sharp decline in abundance in relation to its point of introduction, up to a threshold distance of approximately 80 km beyond which T. fuscum is rare in comparison with native T. cinnamopterum. The restricted range of T. fuscum 20 yr after its introduction may be attributed to limited dispersal of adults or reproductive failures of low-density populations. The distribution of T. fuscum seemed stable between 2008 and 2010. In 1 of 3 yr, the abundance of T. cinnamopterum increased with the distance to the site of introduction of T. fuscum, which suggests competitive interactions between the two Tetropium species.     

Evidence for low-titre infections in insect symbiosis: Wolbachia in the bark beetle Pityogenes chalcographus (Coleoptera, Scolytinae)

Wolbachia are obligatory endosymbiotic α-proteobacteria found in many insect species. They are maternally transmitted and often exhibit reproductive phenotypes like cytoplasmic incompatibility. Pityogenes chalcographus is a bark beetle causing severe damage in spruce stands. Its European populations are divided into several mitochondrial clades separated by partial crossing barriers. In this study, we tested a large sample set covering the natural range of the beetle in Europe for the presence of Wolbachia and associations between infection pattern and mitotypes using a highly sensitive nested PCR technique. 35.5% of the individuals were infected with the endosymbiont and two distinct strains were identified. Both strains occur in low titre not accessible by conventional detection methods. The infections are present all over Europe, unlikely to cause the partial crossing barriers in this host and uncoupled from mitochondrial clades. This pattern is indicative for populations evolving towards endosymbiont loss and for repeated intraspecific horizontal transfer of Wolbachia . Alternatively, the low-titre infections found in P. chalcographus are yet another example for Wolbachia that can persist in host species at low densities and frequencies.     

Ploidy of the eusocial beetle Austroplatypus incompertus (Schedl) (Coleoptera, Curculionidae) and implications for the evolution of eusociality

In the hymenopterans, haplodiploidy, leading to high-genetic relatedness amongst full sisters has been regarded as critical to kin selection and inclusive fitness hypotheses that explain the evolution of eusociality and altruistic behaviours. Recent evidence for independent origins of eusociality in phylogenetically diverse taxa has led to the controversy regarding the general importance of relatedness to eusociality and its evolution. Here, we developed a highly polymorphic microsatellite marker to test whether the eusocial ambrosia beetle Austroplatypus incompertus (Schedl) is haplodiploid or diplodiploid. We found that both males and females of A. incompertus are diploid, signifying that altruistic behaviour resulting from relatedness asymmetries did not play a role in the evolution of eusocialty in this species. This provides additional evidence against the haplodiploidy hypothesis and implicates alternative hypotheses for the evolution of eusociality.     

Fitness consequences of malathion-specific resistance in red flour beetle (Coleoptera: Tenebrionidae) and selection for resistance in the absence of malathion

Malathion resistance in the red flour beetle, Tribolium castaneum (Herbst), is a worldwide problem and is very stable once it becomes widespread in natural populations. In the absence of insecticide the proportion of resistant phenotypes may rapidly decline but the development of resistance does not always involve reduced fitness. Malathion-specific resistance in T. castaneum seems not to involve any loss of fitness in laboratory or field conditions. Susceptible beetles were in competition with resistant beetles at different initial frequencies and modifications of susceptible gene frequency were estimated in these laboratory populations over 10 generations. A significant decrease in susceptible gene frequency was observed in Tribolium populations over time. The selection coefficient of the susceptible allele was estimated and the fitness of susceptible alleles in all tests was observed to range from 0.89 to 0.93 compared with the fitness of resistant genotypes, which was assumed to be 1. Data provided evidence that the resistant strains exhibited fitness advantages in the absence of malathion. We also compared the biotic potential (fecundity and developmental time) of the susceptible strain, the homozygous malathion-specific resistant strain, and their hybrids. Malathion-specific resistant strains showed an 8 -23% increase in biotic potential relative to the susceptible strain. These findings are consistent with those of malathion-specific resistance in T. castaneum; the fitness of the insects seems independent of the genetic background and the fitness of the resistant insects is not affected by this resistance mechanism.     

Lip anatomy and its implications for the pollination mechanisms of Bulbophyllum species (Orchidaceae)

BACKGROUND AND AIMS: The lip structure of six Brazilian and one Asiatic species of Bulbophyllum with wind-assisted fly pollination (B. involutum, B. ipanemense and B. weddellii) and non-wind-assisted fly pollination (B. epiphytum, B. glutinosum, B. regnellii and B. rothschildianum) was studied to investigate the presence of secretory tissues related to these pollination mechanisms. METHODS: The lip study was carried out through scanning electron microscopy (lip surface) and light microscopy (anatomical features). KEY RESULTS: In most of the species studied, the osmophores (odour glands) were located in the lobes and in the upper surface of the lip callus. Differences in the lip structure were observed between the two groups (the presence of a nectary and the extent of osmophore surface), depending on the mechanism of pollination. Nectaries were found in the cavity callus in B. ipanemense, B. involutum and B. weddellii, even though their pollinators were presumably attracted by the instinct to oviposit. CONCLUSIONS: These findings corroborate the hypothesis that, because pollination in these species is dependent on an unpredictable external factor (wind), nectar is necessary to keep the insect in the flower for a long period. Despite the occurrence of a liquid-like nectar in the flowers of B. epiphytum, B. glutinosum, B. regnelli and B. rothschildianum, no anatomical evidence for nectaries was found in the lips of these species, although a similar structure may occur in another region of the flowers. This observation agrees with the fact that pollination by lip movement in the latter species requires only gravity, with no additional mechanism being needed to keep the flies in the flower.     

Photoperiod modifies thermal reaction norms for growth and development in the red poplar leaf beetle Chrysomela populi (Coleoptera: Chrysomelidae)

Regression lines of development rate on temperature appeared significantly different between long (22 h) and short (12 h) day conditions and intersected each other at 23.8 °С. Thus, the rate of growth and development was higher at temperatures below the intersection point under short-day but above the intersection point it was higher under long day. Ecological relevance of this effect seems as follows: in autumn, as nights become longer and average daily temperature decreases, larvae have to speed up their development because it is only imago that overwinters. Conversely, midsummer offers long days and usually higher temperature, so again it is advantageous to develop as fast as possible in order to have at least one more generation per year. These results are compared with other studies showing interactions between photoperiod and temperature, and some possible general patterns are outlined. The lower thermal threshold for larval development depended on photoperiodic conditions; therefore rate isomorphy must be violated in this species. Development at higher temperatures generally resulted in smaller adults, as is usual with ectotherms according to the “temperature-size rule”, but body weight depended significantly on temperature only under short day. Our estimates of the lower temperature thresholds for growth and development in both cases did conform to the generalization made previously by Walters and Hassall (2006) in spite of another formula used by us. We briefly discuss this phenomenon and argue that relative position of these thresholds can be explained mathematically and per se may lack any biological sense.     

Carabid beetle (Coleoptera: Carabidae) conservation in New Zealand

An overview of carabid beetles in the New Zealand conservation context is provided. Processes for threatened species recovery within the New Zealand Department of Conservation are outlined, and the Department’s two major beetle-related recovery documents (The Conservation Requirements for New Zealand’s Nationally Threatened Invertebrates, and The Carabid Beetle Recovery Plan) are discussed. A subjective consideration of the merits and drawbacks of both the documents, and the systems that support them, is presented, along with an option for the way forward for threatened species conservation in New Zealand.     

The maintenance of extensively exploited pastures within the Alpine mountain belt: implications for dung beetle conservation (Coleoptera: Scarabaeoidea)

The abandonment of marginal areas and the decline of traditional small-scale cattle-breeding is bringing about substantial changes in many areas of the Alpine mountain belt. Extensively exploited pastures, which were colonized by typical ecological communities, are now being replaced by shrublands and forests. In this paper, we evaluate the effectiveness of the maintenance of small pastures for dung beetle (Scarabaeoidea: Geotrupidae, Aphodiidae, Scarabaeidae) conservation in a protected area of the north-western Italian Alps. From July until September 2007, we studied the dung beetle coenosis in the main habitats of the mountain belt: beech forest, scots pine forest, mountain pine forest and pasture. Twenty-three species were sampled. We compared abundance, species richness, α-diversity, evenness and species turnover between the four habitat assemblages and performed a correspondence analysis based on dominance patterns of the three families. Two main ensembles were identified: a pasture ensemble, highly diverse and dominated by Scarabaeidae, and a closed habitats ensemble, less structured and dominated by Aphodiidae and Geotrupidae. Combining habitat specificity and fidelity, the IndVal method showed that half of the species collected made a strong habitat selection toward pasture, while only C. granarius (Aphodiidae) was a reliable indicator of woody areas. We therefore conclude that the loss of extensively exploited pastures may bring about fundamental changes in dung insect communities. In protected areas, a management policy intended to preserve the traditional, sustainable human activities within the mountain belt is highly recommended for the conservation of these insects.     

Compensatory proteolytic responses to dietary proteinase inhibitors in the red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae)

Increasing levels of inhibitors that target cysteine and/or serine proteinases were fed to Tribolium castaneum larvae, and the properties of digestive proteinases were compared in vitro. Cysteine proteinases were the major digestive proteinase class in control larvae, and serine proteinase activity was minor. Dietary serine proteinase inhibitors had minimal effects on either the developmental time or proteolytic activity of T. castaneum larvae. However, when larvae ingested cysteine proteinase inhibitors, there was a dramatic shift from primarily cysteine proteinases to serine proteinases in the proteinase profile of the midgut. Moreover, a combination of cysteine and serine proteinase inhibitors in the diet prevented this shift from cysteine proteinase-based digestion to serine proteinase-based digestion, and there was a corresponding substantial retardation in growth. These data suggest that the synergistic inhibitory effect of a combination of cysteine and serine proteinase inhibitors in the diet of T. castaneum larvae on midgut proteolytic activity and beetle developmental time is achieved through the prevention of the adaptive proteolytic response to overcome the activity of either type of inhibitor.     

Dung beetle (Coleoptera: Scarabaeinae) persistence in Amazonian forest fragments and adjacent pastures: biogeographic implications for alpha and beta diversity

The aim of this study is to analyze the effects of habitat loss and forest replacement by cattle pasture on the alpha and beta diversity, abundance, biomass and species composition of dung beetles with different dispersal ability. Dung beetles were captured in 19 forest fragments and neighbouring pastures. Forest fragment area ranged from 3.7 to 4825 ha and in this study were grouped into four categories: small, medium, large and control forest. A total of 35,048 dung beetles representing 101 species were collected. Forest fragments had the highest richness with 81 species, followed by pasture with 58 species. Replacement of forest by pasture reduced species richness; however, due to the proximity and connectivity of these areas with Cerrado patches, pastures also had high species richness, but species composition was independent of adjacent fragments. Small fragments had lower abundance and species richness than our other habitat categories, even pastures. Our results highlight that proximity and connectivity with Cerrado areas influenced the patterns of alpha and beta diversity of dung beetles in fragments and pastures. We highlight that the ability to cross the pasture matrix is a strong adaptive trait for species living in human-modified landscapes. Consequently, species with these abilities are less susceptible to the effects of forest fragmentation and local extinction. Our results reinforce the importance of considering the biogeographic location and distribution pattern of species in forest fragmentation studies.     

Aeration management for stored hard red winter wheat: simulated impact on rusty grain beetle (Coleoptera: Cucujidae) populations

Simulation studies were conducted to determine temperature accumulations below defined thresholds and to show the impact of controlled aeration on populations of the rusty grain beetle, Cryptolestes ferrigineus (Stephens), a major secondary pest of stored wheat, Triticum aestivum (L.). Recorded data from weather stations in Texas, Oklahoma, Kansas, eastern New Mexico, and eastern Colorado (356 total) were used to determine hours of temperature accumulation below 23.9 degrees C in June and July, 15.6 degrees C in September and October, and 7.2 degrees C in December. At an airflow rate of 0.0013 m3/s/m3 (0.1 cubic ft3/min/bu), which requires 120 h of temperatures below the specified threshold to complete an aeration cycle, summer cooling at 23.9 degrees C in bulk-stored wheat could be completed throughout the hard red winter wheat zone except for extreme southern Texas. An early-autumn cooling cycle at 15.6 degrees C could not be completed throughout most of Texas and Oklahoma before the end of September. The late-autumn cooling cycle could be completed in all states except Texas by the end of November. Five geographic regions were delineated and the times required for completion of the summer, early-autumn, and late-autumn cooling cycles within each region were estimated. Population growth of the rusty grain beetle was modeled for San Antonio, TX; Abilene, TX; Tulsa, OK; Topeka KS; and Goodland, KS, by predicting the numbers of adults in the top, outer middle, outer periphery, and the center of the bin during a 1-yr storage season. Populations of C. ferrugineus in San Antonio and Austin were predicted to exceed the Federal Grain Inspection Service (FGIS) threshold of two beetles per kilogram of wheat in all four levels of the bin during late autumn, decline during the winter, and increase the following spring. In Midland, TX, and Oklahoma City, OK, populations were predicted to exceed the threshold only in the top and outer middle of the bin, whereas populations in the Kansas locations were not predicted to exceed the threshold at any time.     

Spermatophore expulsion in the carrion beetle Silpha perforata (Coleoptera: Silphidae)

Silphinae (Coleoptera: Silphidae) is an abundant decomposer that plays important roles in the ecosystem. However, there is little information about the life history of this taxon. We found sperm displacement behavior in carrion beetle Silpha perforata. Copulating males bit the female's antenna strongly and inserted the penis into the partner's genital organ more than once. We found a white substance on the tip of penis during copulation. We examined whether this white substance is a previous male's spermatophore, which was removed from the mating partner. When females were dissected just after mating, the same substance that often presents on the penis of mating males was found in the bursa copulatrix of females, although the bursa copulatrix of virgin females was empty. Male behavior during copulation with females of different mating history was also observed to confirm that the removal of spermatophores was observed only in copulation with females that have the spermatophores of previous males. Consequently, we estimated that S. perforata males removed spermatophores of previous males from mating partners. In addition, we dissected the males frozen during copulation, and inspected the penis morphology. This observation revealed that the apical part of the penis was usually hidden in the basal part of penis, but expanded and appeared during insertion. This apical part had many spines, which play an important role in sperm displacement and sexual conflict in some species. These results indicate that there is the sperm competition in S. perforata. This is the first report on sperm competition in Silphinae.     

Suitability of exotic and native lady beetle eggs (Coleoptera: Coccinellidae) for development of lady beetle larvae

Predation upon lady beetle (Coleoptera: Coccinellidae) eggs in the field is most often instances of egg cannibalism by larvae or adults while the majority of the remaining predation events upon coccinellid eggs is done by other species of Coccinellidae. Thus the recent introduction and establishment of Harmonia axyridis in the US could negatively affect native species of Coccinellidae via egg predation. However, little is known regarding the suitability of interspecific coccinellid eggs as a food source for larval development. In this study, it was found that native first or third instar Coleomegilla maculata and Olla v-nigrum larvae were incapable of surviving to the adult stage when provided solely exotic H. axyridis eggs. In stark contrast, H. axyridis larvae survived equally well when cannibalizing eggs or eating eggs of either native species. When C. maculata and O. v-nigrum were grouped as ‘native’ and compared with the exotic H. axyridis, more native eggs were attacked than exotic eggs and a higher percentage of eggs was attacked by H. axyridis larvae. Native and exotic larvae attacked a similar percentage of native eggs but native larvae attacked significantly fewer exotic eggs than did exotic larvae. These data suggest that H. axyridis may prey upon the eggs of these native species, when encountered in the field, compared with the likelihood of the native species preying upon H. axyridis eggs. Therefore, eggs of the native species C. maculata and O. v-nigrum will continue to be subjected to cannibalism and also to possible predation by other native species and the exotic H. axyridis.     

Complete mitochondrial genome of the darkling beetle Gonocephalum outreyi (Coleoptera: Tenebrionidae) with phylogenetic implications

The complete mitochondrial genome (mitogenome) of Gonocephalum outreyi was determined by using next-generation sequencing approach. The full length of this mitogenome is 15,836?bp, which consists of 37 typical metazoan mitochondrial genes with an identical genome organization to ancestral insects. The majority of the protein-coding genes begin with the codon ATN, except for cox1 and cox2 with AAT and AAA, respectively. To elucidate the phylogenetic position of G. outreyi, we used various sequence coding schemes for protein-coding genes and the combined nucleotide sequences of all mitochondrial genes for tree building under the Bayesian and Maximum Likelihood inferences. The phylogenetic results consistently supported G. outreyi as a member of the family Tenebrionidae. The monophyly of both Tenebrionoidea and Tenebrionidae were strongly supported. The Scraptiidae and Melandryidae were recovered to be non-monophyletic in regards to the Osphya. Within Tenebrionidae, the subfamilies Diaperinae and Tenebrioninae were found to be non-monophyletic.