Benzoquinones and Hydroquinones in Defensive Secretions of Tropical Millipedes

 The defensive secretions of two tropical species of millipedes (the spirostreptid Telodeinopus aoutii and a species of Harpagophoridae) contain a complex mixture of closely related benzoquinones and hydroquinones. The major compounds are toluquinone and 2-methoxy-3-methylbenzoquinone, accompanied by the minor components, 2,3-dimethoxybenzoquinone and toluhydroquinone. Because of the large size and the geographic separation of the test animals a common defensive strategy of tropical, large millipedes against predation by vertebrates is assumed. Received: 30 August 1999 / Accepted in revised form: 20 October 1999     

Benzoquinones from millipedes deter mosquitoes and elicit self-anointing in capuchin monkeys (Cebus spp.)

Neotropical monkeys of the genus Cebus anoint themselves by rubbing arthropods and plants against their pelage. A recent study has shown that free-ranging wedge-capped capuchin monkeys ( C. olivaceus) in Venezuela self-anoint with a benzoquinone-secreting millipede, an activity by which they are hypothesized to appropriate chemical deterrents of mosquitoes. To evaluate the plausibility of this hypothesis, female yellow fever mosquitoes ( Aedes aegypti) were presented with two millipede secretory compounds, 2-methyl-1,4-benzoquinone and 2-methoxy-3-methyl-1,4-benzoquinone, on nylon-reinforced silicone membranes placed over wells filled with human blood, a highly preferred food. Mosquitoes exhibited fewer landings, fed less frequently, and flew more frequently (a possible indication of repellency) in the presence of membranes treated with benzoquinones than with controls. These compounds also elicit self-anointing in captive male and female tufted ( C. apella) and white-faced ( C. capucinus) capuchin monkeys.     

Orange/lemon-scented beetles: opposite enantiomers of limonene as major constituents in the defensive secretion of related carabids

The major constituent in the pygidial gland defensive fluid of the carabid beetle Ardistomis schaumii is (R)-(+)-limonene, whereas that of Semiardistomis puncticollis is (S)-(−)-limonene. This was an unanticipated result, since it is not very common to find the opposite enantiomers of the same compound among the secondary metabolites of related species. Moreover, the glandular liquid of A. schaumii contains 1,8-cineole, and that of S. puncticollis has β-pinene, β-phellandrene, sabinene, and p-cymene. Of about 500 carabid species that have been chemically investigated, this is the first report of the presence of such complex mixtures of monoterpenes in their defensive secretions.     

Hydrogen peroxide as a new defensive compound in “benzoyl cyanide” producing polydesmid millipedes

Hydrogen peroxide was newly and simultaneously demonstrated with well-known hydrogen cyanide as a component of defensive secretions of “benzoyl cyanide” producing polydesmid millipedes. Presence of hydrogen peroxide was successively evidenced by Trinder reagent’s spray with colorless as well as oily smears of defensive secretions containing benzoyl cyanide and hydrogen cyanide by alkaline picrate paper treatment. Linear correlation was demonstrated between quantities of hydrogen peroxide and benzoyl cyanide. By qualitative assay, seven benzoyl cyanide containing polydesmidans (six species of adults and one species of a nymph at stadium I) tested positive to Trinder reagent, indicative of the presence of hydrogen peroxide (together with hydrogen cyanide), while two cyanogenic species without benzoyl cyanide exhibited negative responses to the reagent. Two types of millipedes were elucidated as species of cyanogenic Polydesmida.     

Triterpene saponin hemi-biosynthesis of a leaf beetle’s (<Emphasis Type="Italic">Platyphora kollari</Emphasis>) defensive secretion

The adults of the leaf beetle Platyphora kollari (Chrysomelidae) are able to metabolise the oleanane triterpene β-amyrin (1) into the glycoside 3-O-β-d-glucopyranosyl-(1→4)-β-d-glucuronopyranosyl-hederagenin (2) that is stored in their defensive glands. The aim of this study was to test the hypothesis that oleanolic acid (3) is an intermediate in the conversion of 1 into 2 and to check whether the sequestration of pentacyclic triterpenes is selective in favour of β-amyrin (1). To this end, adults of P. kollari were fed with Ipomoea batatas leaf disks painted with a solution of [2,2,3-²H₃]oleanolic acid or [2,2,3-²H₃]α-amyrin and the secretion of their defensive glands analysed by HPLC–ESIMS. The data presented in this work indicated that the first step of the transformation of β-amyrin (1) into the sequestered glycoside 2 is its oxidation into oleanolic acid (3) and that this conversion is selective but not specific in favour of β-amyrin (1).     

A trail pheromone component of the ant Mayriella overbecki Viehmeyer (Formicidae: Myrmicinae)

 The myrmicine ant Mayriella overbecki lays recruitment trails during foraging and nest emigrations. The trail pheromone originates from the poison gland. From ten identified components of the poison gland secretions only methyl 6-methylsalicylate 1 elicited trail following behavior. Received: 6 April 2000 / Accepted in revised form: 6 June 2000     

Behavioral genomics of honeybee foraging and nest defense

The honeybee has been the most important insect species for study of social behavior. The recently released draft genomic sequence for the bee will accelerate honeybee behavioral genetics. Although we lack sufficient tools to manipulate this genome easily, quantitative trait loci (QTLs) that influence natural variation in behavior have been identified and tested for their effects on correlated behavioral traits. We review what is known about the genetics and physiology of two behavioral traits in honeybees, foraging specialization (pollen versus nectar), and defensive behavior, and present evidence that map-based cloning of genes is more feasible in the bee than in other metazoans. We also present bioinformatic analyses of candidate genes within QTL confidence intervals (CIs). The high recombination rate of the bee made it possible to narrow the search to regions containing only 17–61 predicted peptides for each QTL, although CIs covered large genetic distances. Knowledge of correlated behavioral traits, comparative bioinformatics, and expression assays facilitated evaluation of candidate genes. An overrepresentation of genes involved in ovarian development and insulin-like signaling components within pollen foraging QTL regions suggests that an ancestral reproductive gene network was co-opted during the evolution of foraging specialization. The major QTL influencing defensive/aggressive behavior contains orthologs of genes involved in central nervous system activity and neurogenesis. Candidates at the other two defensive-behavior QTLs include modulators of sensory signaling (Am5HT ₇ serotonin receptor, AmArr4 arrestin, and GABA-B-R1 receptor). These studies are the first step in linking natural variation in honeybee social behavior to the identification of underlying genes.     

Fever in honeybee colonies

 Honeybees, Apis spp., maintain elevated temperatures inside their nests to accelerate brood development and to facilitate defense against predators. We present an additional defensive function of elevating nest temperature: honeybees generate a brood-comb fever in response to colonial infection by the heat-sensitive pathogen Ascosphaera apis. This response occurs before larvae are killed, suggesting that either honeybee workers detect the infection before symptoms are visible, or that larvae communicate the ingestion of the pathogen. This response is a striking example of convergent evolution between this "superorganism" and other fever-producing animals. Received: 2 September 1999 / Accepted in revised form: 28 February 2000     

Heterospecific pairing and hybridization between <Emphasis Type="Italic">Nasutitermes corniger</Emphasis> and <Emphasis Type="Italic">N. ephratae</Emphasis>

The sympatric neotropical termites Nasutitermes corniger and Nasutitermes ephratae are clearly distinguishable based on morphology, nest architecture, defensive secretion composition, and molecular markers. However, given the extensive ecological, geographical, and behavioral overlap of these closely related species, the potential for interbreeding may exist. To explore this possibility, heterospecific pairs were formed experimentally to examine courtship and colony-establishment behaviors, and reproductive potential. Courtship and nest construction behavior occurred in heterospecific pairs in a similar manner to that of conspecific pairs. Survival of pairs depended upon the species of the female partner. N. ephratae females paired with N. corniger males produced as many offspring as conspecific pairs. N. corniger females mated to N. ephratae males, however, produced significantly fewer offspring at 60 days post-establishment than the reciprocal cross or conspecific N. ephratae or N. corniger pairs. This was also the only pairing in which any aggression was observed. Heterospecific pairs and groups formed in mate choice mesocosms, suggesting that species recognition between these two termites is not an important aspect of mate choice. Overall, species mismatch tolerance and hybrid offspring viability are high. The present data, together with previous evidence from defensive secretions and isozyme analysis, suggest that hybridization may periodically occur in nature, and that reproductive barriers between these two species may be incomplete. Hybridization could provide a rare but important source of genetic diversity and may ensure mating opportunities for the more abundant sex of alates in each species.     

2,3-Dimethyl-5-(2-methylpropyl)pyrazine, a trail pheromone component of Eutetramorium mocquerysi Emery (1899) (Hymenoptera: Formicidae)

 The ant Eutetramorium mocquerysi (Myrmicinae) is endemic to the island of Madagascar. During foraging and nest emigration the ants lay recruitment trails with secretions from the poison gland. We identified three pyrazine compounds in the poison gland secretion: 2,3-dimethyl-5-(2-methylpropyl)pyrazine 1, 2,3-dimethyl-5-(3-methylbutyl)pyrazine 3, 2,3-dimethyl-5-(2-methylbutyl)pyrazine 4. Only the first component elicited trail-following behavior in the ants. We were unable to investigate whether the other pyrazine components have a synergistic function. Received: 21 February 2000 / Accepted in revised form: 27 June 2000     

Mycotoxin production on rice, pulses and oilseeds

 Mycotoxin-producing fungi were isolated from contaminated grains of rice, pulses and oilseeds sold in the local markets of Calcutta for human consumption. It was found that aflatoxin B₁ was produced by Aspergillus flavus and Aspergillus parasiticus, aflatoxin G₁ by A. flavus, ochratoxin by Aspergillus ochraceous, sterigmatocystin by Aspergillus japonicus and citrinin by Penicillium citrinum. Aflatoxin B₁ (333–10416 μg/kg) was produced by Aspergillus spp. in rice, pulses and oilseeds. Received: 11 May 1999 / Accepted in revised form: 10 April 2000     

Impacts of Climate Change on the Distribution of Larix Spp. and Pinus Sylvestris and Their Climatypes in Siberia

Inter- and intraspecific effects of climate change were assessed for the dominant conifers of Siberia (60–140^∘E and 48–75^∘N): Larix spp. (L. sibirica, L. dahurica, and L. sukaczewii) and Pinus sylvestris . The approach employed a tri-variate (degree-days above 5 ^∘C, degree-days below 0 ^∘C, and a moisture index) estimate of the climatic envelope within which exists the actual ecological distribution of a species and their constituent climatypes (genotypes physiologically attuned to similar environments). Limits of the actual ecological distribution were approximated by reducing the climatic envelope according to effects of permafrost and interspecific competition. Climatypes were mapped within the climatic envelope according to the climatic interval that must separate populations for reasonable assurance of genetic differentiation. This interval was calculated from response functions that related 13-year growth and survival of a species to the difference in climate between the provenance of a climatype and the climate of numerous test sites distributed across Russia. Mapping species' distributions and their climatypes was done for the contemporary climate and for future climates predicted by the HadCM3GGa1 scenario of Hadley Centre. The results showed that if the forests of the future are to reflect the adaptedness of today, the distribution of species will shift and genotypes within species will be redistributed. Some contemporary climatypes are projected to disappear from Siberia while others common elsewhere would evolve. To mitigate these effects, climatypes should be transferred today to the expected future location of their climatic optima, a distance that is likely to approach 700–1200 km for these species.     

The larval parasitoid <Emphasis Type="Italic">Microplitis croceipes</Emphasis> oviposits in conspecific adults

Microplitis croceipes (Hymenoptera: Braconidae) is a larval parasitoid of Helicoverpa/Heliothis spp. In the course of mass rearing of M. croceipes, we found that females oviposited in the conspecific adults in rearing cages. When 20 pairs of inexperienced females and males or of experienced females and males were reared in a cage, the males lived for 14–15 days and the females for 18–20 days on average. At their death, 37–42% of the males and 50–57% of the females contained conspecific eggs or first instar larvae in their abdominal cavity. When two of inexperienced females met on a host-infested leaf of soybean, they attempted to sting each other. Of the attacked females, 30% contained a conspecific egg laid in their abdomen. In abdominal cavity of the adults parasitized by a conspecific female, the majority of the parasitoid eggs laid disappeared within 1 day after oviposition. Only 10–30% of the parasitoid eggs laid in conspecific adults hatched 3–4 days after oviposition, but those larvae never molted to second instar. When the adults were stung by one or two conspecific females, their subsequent longevity was significantly shorter than that for the control adults. Oviposition in conspecific adults may be prevalent in other parasitic wasps that quickly oviposit without intensive host examination, and have cuticle and size of abdomen to be stung by conspeicifcs.     

Recycling plant wax constituents for chemical defense: hemi-biosynthesis of triterpene saponins from β-amyrin in a leaf beetle

Several species of Doryphorina leaf beetles from Central- and South America produce oleanane triterpene glycosides in their defensive glands. The presence of pentacyclic triterpenes in insects is intriguing since they lack the key enzymes necessary to synthesize these compounds. Since -amyrin is a common constituent of leaf waxes, we hypothesized that these leaf beetles use this compound as a precursor to their oleanane glycosides. To test this hypothesis we first confirmed the presence of -amyrin in Ipomoea batatas, the food plant of Platyphora kollari. Next, adults of P. kollari were fed for 10 days with I. batatas leaf disks painted with a solution of [2,2,3-²H₃]-amyrin ([2,2,3-²H₃]-1). The secretion from their defensive glands was collected and analyzed by HPLC-ESIMS. The results demonstrated that the secretion of beetles fed with an amount of [2,2,3-²H₃]-amyrin corresponding to the quantity of unlabeled (natural) -amyrin present in the leaf disks contained on average 5.1% of [2,2,3-²H₃]-3-O--d-glucopyranosyl-(1-->4)--d-glucuronopyranosyl-hederagenin ([2,2,3-²H₃]-2), whereas the secretions of beetles fed with 10 times this amount of [2,2,3-²H₃]-amyrin contained on average 23.9% of [2,2,3-²H₃]-2. In both series of experiments, the percentage of labeled versus unlabeled triterpene glycoside in the secretion was positively correlated with the amount of deuterated -amyrin ingested. These results demonstrate for the first time that some leaf beetles are able to metabolize a widespread triterpenic constituent of leaf wax into more complex glycosides that are stored in their defensive glands.     

How to uncoil your partner—“mating songs” in giant pill-millipedes (Diplopoda: Sphaerotheriida)

The stridulation of the giant pill-millipede genus Sphaerotherium from South Africa, one of only three groups of millipedes that produce sounds, was studied. One hundred one stridulation series of a total of nine different species (Sphaerotherium dorsaloide, Sphaerotherium hanstroemi, Sphaerotherium mahaium, Sphaerotherium similare, Sphaerotherium punctulatum, Sphaerotherium convexitarsum, Sphaerotherium dorsale, Sphaerotherium rotundatum, and Sphaerotherium perbrincki) were analyzed. Stridulation sounds are produced only with a special field of ribs on the posterior surface of the posterior telopod, which is actively moved over a field of sclerotized nubs on the inner margin of the anal shield. The Sphaerotherium male usually stridulates only when in contact with a female to initiate mating. This seems to prevent the female from volvating into a ball or stimulate the female to uncoil when already rolled in. The sound analyzes revealed a broad frequency spectrum in all stridulation sounds produced, without obvious differences in frequency distribution among species. However, the temporal pattern of the stridulation varies greatly between species and seems to be species-specific, arguing for a species recognition function of the stridulation during courtship behavior. A single species (S. punctulatum) was found to stridulate during mating while three species also show postcopulatory stridulation. Apparently, pill-millipedes are not capable of acoustic perception, as no hearing organs are known, indicating that the communication is mainly based on perception of temporal vibration patterns, and not of the acoustic signal itself. The need to overcome the rolling-in reflex of the female is developed as a hypothesis why stridulation exists only in millipedes able to coil into a ball, and apparently evolved four times independently in the superorder Oniscomorpha.     

Size-correlated division of labour and spatial distribution of workers in the driver ant,<Emphasis Type="Italic"> Dorylus molestus</Emphasis>

Driver ants ( Dorylus spp.) show a high degree of worker polymorphism. Previous reports suggest that large Dorylus workers are specialised for defensive tasks. In this study, we first quantitatively tested whether there is a size-correlated division of defensive labour among workers. Second, we determined whether the spatial distribution of workers outside the nest can be predicted based on such size-specific differences in task allocation. We show that the division of defensive behaviour among different-sized workers is not strict. However, there is a significant correlation between worker size and the tendency to carry out defensive tasks. First, workers of larger size were more likely than smaller workers to participate in colony defence. Second, larger workers were more frequent near the nest containing the reproducing individuals and the brood. Finally, large workers were more common in open sections of the trail than in covered sections, which are likely to be less exposed to predators.     

What’s the buzz? Ultrasonic and sonic warning signals in caterpillars of the great peacock moth (Saturnia pyri)

Caterpillars have many natural enemies and, therefore, have evolved a diversity of antipredator strategies. Most research focuses on those strategies (crypsis, countershading, and warning coloration) targeting visually guided predators. In contrast, defensive sounds, although documented for more than a century, have been poorly studied. We report on a novel form of sound production—chirping—in caterpillars of the common European Great Peacock moth (Saturnia pyri). Chirps are broadband, with dominant peaks ranging between the sonic (3.7 kHz) and ultrasonic (55.1 kHz) and are generated by a rapid succession of mandibular “tooth strikes.” Chirp trains are induced by simulated predator attacks and precede or accompany the secretion of a defensive chemical from integumental bristles, supporting our hypothesis that these sounds function in acoustic aposematism. We propose that these caterpillars generate multimodal warning signals (visual, chemical, and acoustic) to target the dominant sensory modalities of different predators, including birds, bats, and invertebrates. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

新安江水库河口区水质及藻类群落结构高频变化

水库库尾区的水环境多变,是水库生态系统突变的重要策源地.为探究大型水库水源地水环境演变特征及其突变的促发机制,以新安江水库为例,通过库尾河口断面18个月水质浮标的高频记录及3 d一次的藻类群落结构人工鉴定数据等,分析了气象水文过程影响下的水库库尾区的水温、溶解氧、浊度及营养盐等环境指标及藻类群落结构的高频变化特征,揭示了降雨、入流及季节温度变化等关键气象水文过程对水库水质及藻类群落结构的影响机制.结果表明:①在27 m深的河流入库区的水体温度和溶解氧存在明显的季节分层,相应水体藻类叶绿素a和营养盐等指标也同步发生分层,水温分层从气温达到14℃以上的3月中旬开始,至气温降至24℃后的10月中旬结束,期间较大降雨和入流多次破坏水温分层;②河道入库区水体氮、磷等营养盐变幅大,总磷浓度变幅为0. 011～0. 188 mg·L~(-1)之间,总氮浓度变幅为0. 75～2. 76 mg·L~(-1)之间,总磷和总氮中的溶解态占比分别为56%及88%,降雨入流对水体营养盐浓度影响巨大,3 d的累积降雨与水体氮、磷浓度显著正相关,3～6月(雨季)的营养盐含量明显高于其他月份(P 0. 001),藻类的季节性增殖反过来也会影响水体总磷浓度;③藻类群落结构及其优势属呈现明显的季节变化,在总体硅藻门类占优的背景下,蓝藻、绿藻、隐藻等在不同季节形成明显峰值,蓝藻在7～10月的夏秋季形成明显的生物量峰值,其峰值形成原因除了高温之外,还与暴雨入流有关.蓝藻主要优势属为束丝藻属(Aphanizomenon spp.)、微囊藻(Microcystis spp.)及颤藻(Oscillatoria spp.)等,绿藻峰值与蓝藻基本同步,优势属为盘星藻属(Pediastrum spp.)和新月藻属(Closterium spp.),隐藻在3～5月形成峰值,优势属为隐藻属(Cryptomonas spp.),硅藻门中的优势属分别为脆杆藻属(Fragilaria spp.)、小环藻属(Cyclotella spp.)、针杆藻属(Synedra spp.)及直链藻属(Melosira spp.)等;④入库流量、温度、水位、透明度、总氮、总磷及氮磷比等均为影响藻类优势属演替的主要因子,秋冬季节的控制因子为气象水文条件,而夏秋季节则受气象水文及营养盐的共同控制.本研究表明强降雨过程能对水库库尾区水环境及水生态系统结构产生巨大冲击,是水库藻类水华发生的可能诱发因子,通过对该过程的规律认识及关键指标监测,能够为水库水源地水质风险提供预警信息.     

Cerumen of Australian stingless bees (<Emphasis Type="Italic">Tetragonula carbonaria</Emphasis>): gas chromatography-mass spectrometry fingerprints and potential anti-inflammatory properties

Cerumen, or propolis, is a mixture of plant resins enriched with bee secretions. In Australia, stingless bees are important pollinators that use cerumen for nest construction and possibly for colony’s health. While extensive research attests to the therapeutic properties of honeybee (Apis mellifera) propolis, the biological and medicinal properties of Australian stingless bee cerumen are largely unknown. In this study, the chemical and biological properties of polar extracts of cerumen from Tetragonula carbonaria in South East Queensland, Australia were investigated using gas chromatography-mass spectrometry (GC-MS) analyses and in vitro 5-lipoxygenase (5-LOX) cell-free assays. Extracts were tested against comparative (commercial tincture of A. mellifera propolis) and positive controls (Trolox and gallic acid). Distinct GC-MS fingerprints of a mixed diterpenic profile typical of native bee cerumen were obtained with pimaric acid (6.31 ± 0.97%, w/w), isopimaric acid (12.23 ± 3.03%, w/w), and gallic acid (5.79 ± 0.81%, w/w) tentatively identified as useful chemical markers. Characteristic flavonoids and prenylated phenolics found in honeybee propolis were absent. Cerumen extracts from T. carbonaria inhibited activity of 5-LOX, an enzyme known to catalyse production of proinflammatory mediators (IC₅₀ 19.97 ± 2.67 μg/ml, mean ± SEM, n = 4). Extracts had similar potency to Trolox (IC₅₀ 12.78 ± 1.82 μg/ml), but were less potent than honeybee propolis (IC₅₀ 5.90 ± 0.62 μg/ml) or gallic acid (IC₅₀ 5.62 ± 0.35 μg/ml, P < 0.001). These findings warrant further investigation of the ecological and medicinal properties of this stingless bee cerumen, which may herald a commercial potential for the Australian beekeeping industry.