Identification of the alkaloidal venoms of some Monomorium ants of Saudi Arabia

The major volatile compounds in the poison glands of two Monomorium ant species from Saudi Arabia have been identified. Monomorium niloticum and Monomorium najrane both contain mixtures of alkyl- and alkenyl-pyrrolidines and -pyrrolines in their venom glands but no Dufour gland volatile compounds were detected. Monomorium mayri showed neither Dufour gland compounds nor venom components detectable by gas chromatography.     

Novel diterpenoids and hydrocarbons in the Dufour gland of the ectoparasitoid Habrobracon hebetor (Say) (Hymenoptera: Braconidae)

Chemical constituents contained in the Dufour gland of the ectoparasitoid Habrobracon hebetor (Say) (Hymenoptera: Braconidae) were characterized. Three terpenes, beta-springene, a homo-beta-springene, and a homo-geranyllinalool constitute approximately 37% of the gland components, with the remaining 63% all being hydrocarbons. The hydrocarbons consist of a homologous series of n-alkanes (n-C21 to n-C31), a trace amount of 3-methyl C23, a homologous series of internally methyl-branched alkanes (11-methyl C23 to 13-methyl C35), one dimethylalkane (13,17-dimethyl C33), a homologous series of monoenes (C(25:1) to C(37:1)) with the double bonds located at Delta9, Delta13 and Delta15 for alkenes of carbon number 25 to 31 and at Delta13 and Delta15 for carbon numbers 33 to 37 and three homologous dienes in very low amounts with carbon numbers of 31, 32, and 33. The terpenoid and hydrocarbon composition of the Dufour gland was similar in virgin and mated females. However, in contrast to the hydrocarbons, the amount of beta-springene and homo-geranyllinalool increased significantly with time after adult emergence from the cocoon. Although many hydrocarbons in the Dufour gland are the same as those on the cuticle of this species [Howard and Baker, Arch. Insect Biochem. Physiol. 53:1-18 (2003)], substantial differences also occur. Of particular note is the chain length of alkenes and location of the double bonds: cuticular alkenes have a chain length of C23 to C29 and double bond locations at Delta5, Delta7, and Delta9, whereas the Dufour gland alkenes contains a greater range of carbon numbers and have no Delta5 or Delta7 alkenes. The Dufour gland contains only one of the long-chain dimethylalkanes found on the cuticle. Also, no terpenoids are found on the cuticle, and the Dufour gland contains none of the secondary wax esters that are major components on the cuticle. GC-MS analysis of lipids carried in the hemolymph of H. hebetor indicated that all hydrocarbons found on both the cuticle and in the Dufour gland are present, as are some of the wax esters. However, none of the terpenoids were detected in the hemolymph. This suggests that the hydrocarbons are synthesized in other tissues or cells, probably by oenocytes, and differentially partitioned between the cuticle and the Dufour gland. The terpenoids are most likely synthesized within the Dufour gland. Analysis of surface lipids from eggs laid within 18 h indicated that no diterpenoids were present. Rather, the lipids present on the eggs were n-alkanes, monomethylalkanes, alkenes, and secondary alcohol wax esters. This composition did not reflect that of the Dufour gland, hence eggs are not being coated with Dufour gland components during oviposition.     

Chemical constituents of Dufour's gland in the ant, Myrmica rubra

The volatile constituents of the Dufour gland in Myrmica rubra have been examined by gas chromatography-mass spectrometry. The principal constituents are 8-heptadecene, n-pentadecane, n-heptadecane, and 9-nona-dencene. The other constituents comprise mainly other alkanes, alkenes, and a few sesquiterpenes. The sting gland does not contain volatile materials.     

Dufour gland secretions of Myrmica rugulosa and Myrmica schencki workers

The chemical composition and behavioural activities of the secretions of the Dufour glands of Myrmica rugulosa and M. schencki have been studied, as part of an extended study on Myrmica ants. Chemically, the Dufour gland of M. rugulosa is filled with a mixture of hydrocarbons dominated by straight chain alkanes and alkenes with 13 to 19 carbon atoms, as found in M. rubra. Significant quantities of (Z,E)-α-farnesene and its homologues, homofarnesene and bishomofarnesene, are also present. In M. schencki, the major compounds present are homofarnesene and bishomofarnesene. In both species, the very volatile portion of the Dufour gland secretion is identical to that analysed in M. rubra. From an ethological point of view, this very volatile part is efficient in attracting workers at a distance (6 to 8 cm) and in decreasing their wandering movements. No specificity was observed when performing cross-tests with Dufour glands freshly isolated from workers of other Myrmica species, but obvious specificities were detected when testing the less volatile part of the Dufour glands' contents, known to be used for marking newly discovered areas.     

Comparative study of the dufour gland secretions of workers of four species of Myrmica ants

The Dufour gland secretions of myrmica rubra, M. ruginodis, M. sabuleti and M. scabrinodis have been studied. The most volatile portions of the secretion of workers of all four species were found to be similar, containing C₂C₄ oxygenated compounds. The less volatile portion consists of a mixture of hydrocarbons. In M. ruginodis this is chiefly a mixture of linear saturated and mono-unsaturated hydrocarbons, similar in composition to that of M. rubra, while in M. sabuleti it consists of (Z,E)-α-farnesene and its homologues, homofarnesene, bishomofarnesene and trishomofarnesene, similar in composition to that of M. scabrinodis. Workers of each species studied were attracted to the Dufour gland volatiles of all four species, these substances chiefly causing an increase in running speed, with the workers not distinguishing between conspecific and allospecific secretions, though small quantitative differences could be demonstrated between the speed and orientation reaction of workers of each species. The less volatile fraction of the Dufour gland secretion is used for territorial marking by foraging workers. This marking is specific for each species except between M. rubra and M. ruginodis.     

Dufour gland of the digger wasp<Emphasis Type="Italic"> Liris niger:</Emphasis> structure and developmental and biochemical aspects

The tubiform Dufour gland in the digger wasp species Liris niger is about 1.0 mm long ( 0.15 mm). An alternating arrangement of longitudinal and circumferential bundles of striated muscle fibers surrounds the gland. The Dufour gland, together with the venom gland, enters the sting base and terminates in the sting. The glandular epithelium is monolayered. Glands about 3 day after imaginal ecdysis have an empty lumen but a thick lining epithelium. The gland cells are characterized by a well-developed vesicular smooth endoplasmic reticulum, sparse rough ER and numerous free ribosomes. They also exhibit several electron-lucent vesicles and autophagic vacuoles. Secretion of electron-dense material via the gland cuticle into the gland lumen is apparent. Glands more than 20 days after imaginal ecdysis display a large lumen and a thin epithelium. The cells show signs of degeneration with numerous cytolytic inclusions. Dufour gland liquid contains numerous polypeptides of molecular weights ranging from 14 to about 200 kDa. In addition the secretion consists predominantly of straight-chain hydrocarbons, accompanied by small amounts of esters. The major hydrocarbons are pentadecane and (Z)-8-heptadecene. Dufour gland secretion may have several functions: (1) the polypeptides might be involved in the gluing process of the eggs, while (2) the hydrocarbon oils may function as lubricants for the lancets and (3) might soften the secretion, thus allowing easier application of the glue. The lipophilic volatile material (4) might also be involved in pheromonal signaling.     

Dufour gland secretion of Polistes wasp: Chemical composition and possible involvement in nestmate recognition (Hymenoptera: vespidae)

The Dufour gland secretion and cuticle lipids of foundresses of Polistes dominulus have been chemically analysed. The gland secretion (analysed for the first time in a polistine species) contains a complex mixture of hydrocarbons, which are the same as those found on the cuticle. The cuticular hydrocarbon mixture and gland secretion differ, however, in the proportions of components: in the first, linear hydrocarbons are more abundant, while in the second we found a higher quantity of dimethylalkanes. Multivariate statistical analysis has shown that foundresses belonging to different colonies are distinguished on the basis of both the composition of their Dufour gland secretion and cuticular hydrocarbon mixture. Branched hydrocarbons seem to be particularly important in distinguishing females according to their colony. We suggest that the Dufour gland may contribute substances to form the hydrocarbon layer on the cuticle.     

Behavioral and olfactory antennal responses of Solenopsis geminata (Fabricius) (Hymenoptera: Formicidae) workers to their dufour gland secretion

Behavioral and electrophysiological tests were performed to evaluate the responses of workers of the ant Solenopsis geminata (Fabricius) from different size categories to Dufour gland extracts. Morphometric measures based in head widths across eyes were used to determine worker sizes. Trail following response of different worker sizes to Dufour gland extract from workers of different sizes was assessed. For each worker size category olfactory responses to Dufour gland extracts were determined using electroantennography (EAG). Gas chromatography and mass spectrometry (GC-MS) were used to determine the chromatographic profile of Dufour gland secretion for each worker size. Morphometric measures permitted to classify the workers of S. geminata as large, medium and small workers. Medium S. geminata workers displayed a significantly higher behavioral response to Dufour gland extracts produced by medium size workers. Similarly, medium workers showed a significantly higher EAG response to Dufour gland extracts produced by medium sized workers. Chromatographic profile of Dufour gland secretions produced by workers showed that each size category exhibited a characteristic profile of the three main components considered as potential trail pheromone constituents. This work showed that medium workers of S. geminata exhibited a high trail-following behavior as well as a high antennal response to Dufour gland secretion. This and their relative abundance in field foraging areas, suggest that medium-sized workers are specialized in foraging activities.     

Morphology and chemistry of Dufour glands in four ectoparasitoids: Cephalonomia tarsalis,C. waterstoni (Hymenoptera: Bethylidae), Anisopteromalus calandrae,and Pteromalus cerealellae (Hymenoptera: Pteromalidae)

The venom apparatus of four hymenopterous parasitoids, including two bethylids, C. tarsalis (Ashmead) and C. waterstoni (Gahan), and two pteromalids, A. calandrae (Howard) and P. cerealellae (Ashmead), were removed and the associated Dufour glands characterized with respect to their external morphology and chemistry. Dufour glands in all four species have a characteristic translucent appearance that apparently results from their lipid content. The stalked Dufour glands of C. tarsalis and C. waterstoni are pear-shaped and have overall lengths of approximately 0.2 and 0.15 mm, respectively. The thin venom glands are bifurcate and insert through a fine duct into the transparent ovoid- to pear-shaped venom reservoir in these bethylids. In A. calandrae and P. cerealellae the Dufour glands are elongated, tubular structures of ca. 0.35 and 0.8 mm in length, respectively, that constrict to a short stalk that empties into the common oviduct. The venom glands in these pteromalids are simple elongated structures that insert into the sac-like venom reservoir through a fine duct. The chemistry of the volatile contents of the Dufour gland in these four species differs considerably. C. tarsalis Dufour glands contain the same hydrocarbon components as found on the cuticle of this species (Ann. Entomol. Soc. Am. 91:101-112 (1998)), and no other chemicals. The Dufour glands of C. waterstoni also contain only hydrocarbons, most of which are the same as the cuticular hydrocarbons (Ann. Entomol. Soc. Am. 85:317-325 (1992)), but in addition the Dufour gland contains ca. 3% of a mixture of 2,17- and 2,19-dimethyl C(23). A. calandrae Dufour gland chemistry is somewhat more complex than that of either of the two bethylids, but like the bethylids, only hydrocarbons are present. The carbon number range is from C(30) to C(39) and consists of a mixture of n-alkanes (C(30)-C(38)); 3-, 5-, 7-, 9-, 11-, 12-, 13-, 14-, 15- and 17-methyl alkanes; 3,7- and 3,11-dimethyl alkanes; 5,9- and 5,17-dimethyl alkanes; 7,11-, 9,13-, 13,17-, 14,18- and 15,19-dimethyl alkanes; 3,7,11- and 3, 9,15-trimethyl alkanes; and 3,7,11,15-tetramethyl alkanes. The cuticular hydrocarbons of this species have not been previously reported, but they are the same as the Dufour gland hydrocarbons. The Dufour glands of P. cerealellae contain both hydrocarbons and two long-chain aldehydes. Most of the hydrocarbons are identical to those found on the cuticle of this species (Ann. Entomol. Soc. Am. 94:152-158 (2001)), but in addition, 5,9-dimethyl C(27), 5,13-, 5,17- and 5,19-dimethyl C(35), 12- and 14-methyl C(36), 12,16- and 13,17-dimethyl C(36), 13-methyl C(37) and 13,17-dimethyl C(37) are present. The two aldehydes detected in glands from P. cerealellae are n-tetracosanal (C(23)CHO) and n-hexacosanal (C(25)CHO).     

红火蚁工蚁对跟踪信息素的触角电位和行为反应

【目的】测定红火蚁Solenopsis invicta Buren工蚁对其跟踪信息素的触角电位（EAG）及行为反应。【方法】解剖红火蚁工蚁的杜氏腺, 用正己烷溶剂提取其分泌的跟踪信息素进行气相色谱-质谱（GC-MS）分析, 并测定了红火蚁工蚁对杜氏腺提取物、 工蚁提取物和合成的法尼烯混合物的EAG和招募行为反应。【结果】通过与合成的法尼烯混合物的气相色谱（GC）保留时间和质谱图比对, 发现杜氏腺提取物的主要成分并不是Z, E-α-法尼烯。EAG测定结果表明, 红火蚁工蚁对杜氏腺提取物、 工蚁提取物及100 μg的法尼烯混合物均有较强的EAG反应, 其次为10 μg和1 μg的法尼烯混合物。在招募行为测定中, 杜氏腺提取物和工蚁提取物招募作用明显, 而10, 1, 0.1和0.01 μg法尼烯混合物的作用均不显著。【结论】Z, E-α-法尼烯不是红火蚁跟踪信息素的主要成分; 红火蚁工蚁对杜氏腺提取物、 工蚁提取物有较强的EAG反应和明显的招募行为反应。     

Morpho-functional comparison of the Dufour gland in the female castes of the Amazon ant <Emphasis Type="Italic">Polyergus rufescens</Emphasis> (Hymenoptera,Formicidae)

The Dufour gland is crucially involved in main aspects of the parasite habit of the slave-making ant Polyergus rufescens, i.e. slave-raids and host colony usurpation. Workers use chemicals from this gland as recruitment signals during raid organization, while newly-mated queens use its secretion to appease residents during host nest invasion. Here, we report a comparison of the general morphology and fine structure of the Dufour gland in the female castes of P. rufescens: queens, ergatogynes (intermediate forms), and workers. The analysis clearly shows the link between gland structure and its behavioural role in queens and workers. In particular, queens present a hypertrophied gland with an extended lumen and a thin epithelium no more active in secretory function. This is consistent with the fact that usurper queens use the Dufour gland contents only during the short phase of host nest penetration. Contrary to adult queens, the cytoplasmic organization of the Dufour gland epithelium of raiders is typical for a tissue with secretory activity (abundance of mitochondria, free ribosomes, strands of smooth endoplasmic reticulum and a Golgi apparatus). This is consistent with the continuous raiding activity performed by workers throughout their adult life. The biology of ergatogynes is still an enigmatic matter. Their Dufour gland is intermediate in shape and size between that of queens and workers. It presents a fairly thick epithelium with features that are typical of a quite active secretory tissue.     

Structure and chemistry of the Dufour gland in Pristomyrmex ants (Hymenoptera, Formicidae)

Billen, J., Ito, F., Tsuji, K., Schoeters, E., Maile, R. and Morgan, E. D. 2000. Structure and chemistry of the Dufour gland in Pristomyrmex ants (Hymenoptera, Formicidae). —Acta Zoologica (Stockholm) 81 : 159–166 All individuals of the three species of Pristomyrmex studied have a Dufour gland with a conspicuous hammer‐shaped distal part, that is connected to a thin‐walled proximal reservoir through a very narrow stalk. The secretory distal part is formed by high columnar cells that are characterized by a highly folded apical wall. The lateral cell junctions apically correspond with the crenel tops, which gives individual cells a peculiar shape with a deep apical depression. The cytoplasm of the secretory cells contains a very well developed smooth endoplasmic reticulum and Golgi apparatus, numerous mitochondria and lysosomes. Histochemical analysis indicates a positive reaction for the presence of a lipid secretion in the hammer‐like part. Gas chromatographic analysis of glands of P. pungens workers reveals the secretion to be formed of a mixture of simple volatile monoterpene hydrocarbons; α–pinene, β–pinene, limonene and camphene. Similar, but species characteristic mixtures of four monoterpenes were found in Pristomyrmex brevispinosus and Pristomyrmex sp.1. Behavioural experiments did not allow a conclusive determination of the function of the gland.     

Exocrine chemistry of the myrmicine ant Zacryptocerus pusillus (Hymenoptera: Formicidae)

Minor workers of the ant Zacryptocerus pusillus have unusual exocrine secretions in both their mandibular and Dufour glands. The mandibular glands contain a 3:1 mixture of 4-heptanone and 4-heptanol, a mixture found only in the related species Z. varians. The Dufour gland contains a mixture of 13 aldehydes from C9 to C18, not previously encountered in ant secretions. The venom glands gave variable results with only nonanal present consistently.     

四川短尾鼩包皮腺的挥发性化学成分分析

哺乳动物的包皮腺分泌物对个体间性引诱及繁殖行为的信息交流和种群调节有着重要的作用。本研究以四川短尾鼩（Anourosorex squamipes）为研究对象对劳亚食虫目动物包皮腺挥发性化学成分和化学通讯功能进行报道。采用顶空固相微萃取—气质联用（HS-SPME-GC/MS）的方法,分析四川短尾鼩雄体包皮腺中挥发性物质的化学组成。结果表明：（1）四川短尾鼩雄体包皮腺的挥发性化学成分主要含烷烃类、醇类、酮类、醛类、醚类、酯类、酸类、芳香烃类等45种化合物;（2）成年雄性四川短尾鼩的包皮腺中挥发性成分有39种,幼年雄性含有28种,说明四川短尾鼩成年雄体的包皮腺中挥发性化学成分多于幼年雄体;（3）成年雄体的包皮腺分泌物中含有4种特有的挥发性化学成分,幼年雄体中特有成分仅为1种,表明四川短尾鼩包皮腺中的挥发性化学成分存在年龄差异;（4）四川短尾鼩包皮腺分泌物中不同化学成分相对含量不同,同种化学成分在不同个体间的相对含量存在差异。四川短尾鼩雄性包皮腺挥发性化合物种类丰富,个体、年龄差异明显。推测挥发性化合物中丁酸（Butanoicacid）、乙酸乙酯（Ethyl acetate）、苯酚（Phenol）为四川短尾鼩的信息素。本研究为进一步验证该物种的信息素成分及其传递机制提供了基础数据。     

Paralyzing action from a distance in an arboreal African ant species

Due to their prowess in interspecific competition and ability to catch a wide range of arthropod prey (mostly termites with which they are engaged in an evolutionary arms race), ants are recognized as a good model for studying the chemicals involved in defensive and predatory behaviors. Ants' wide diversity of nesting habits and relationships with plants and prey types implies that these chemicals are also very diverse. Using the African myrmicine ant Crematogaster striatula as our focal species, we adopted a three-pronged research approach. We studied the aggressive and predatory behaviors of the ant workers, conducted bioassays on the effect of their Dufour gland contents on termites, and analyzed these contents. (1) The workers defend themselves or eliminate termites by orienting their abdominal tip toward the opponent, stinger protruded. The chemicals emitted, apparently volatile, trigger the recruitment of nestmates situated in the vicinity and act without the stinger having to come into direct contact with the opponent. Whereas alien ants competing with C. striatula for sugary food sources are repelled by this behavior and retreat further and further away, termites defend their nest whatever the danger. They face down C. striatula workers and end up by rolling onto their backs, their legs batting the air. (2) The bioassays showed that the toxicity of the Dufour gland contents acts in a time-dependent manner, leading to the irreversible paralysis, and, ultimately, death of the termites. (3) Gas chromatography-mass spectrometry analyses showed that the Dufour gland contains a mixture of mono- or polyunsaturated long-chain derivatives, bearing functional groups like oxo-alcohols or oxo-acetates. Electrospray ionization-mass spectrometry showed the presence of a molecule of 1584 Da that might be a large, acetylated alkaloid capable of splitting into smaller molecules that could be responsible for the final degree of venom toxicity.     

Morphology and ultrastructure of the Dufour gland in workers of social wasps (Hymenoptera, Vespidae)

The Dufour gland in workers of vespine wasps appears as an unpaired tubiform gland that opens in close proximity to the sting base. The epithelial cells that line the central reservoir are characterized by apical microvillus-like projections and deep basal invaginations. Their cytoplasm contains a well-developed Golgi apparatus, numerous mitochondria, as well as strands of smooth endoplasmic reticulum. The Dufour gland duct occurs ventrally to the venom gland duct, and bends downward near the sting base to open in the dorsal vaginal wall. In this region, the duct is dorsoventrally flattened, and shows conspicuous bundles of parallel microtubules in the epithelial cells, that transmit the pulling forces of the myofilaments of the underlying muscular supply to the cuticle. This results in an active opening mechanism regulated by muscular contraction, while passive closure probably results from the return of the cuticular intima to a rest position.     

Behavioral responses to the alarm pheromone of the ant Camponotus obscuripes (Hymenoptera: Formicidae)

The alarm pheromone of the ant Camponotus obscuripes (Formicinae) was identified and quantified by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Comparisons between alarm pheromone components and extracts from the major exocrine gland of this ant species revealed that the sources of its alarm pheromone are Dufour's gland and the poison gland. Most components of Dufour's gland were saturated hydrocarbons. n-Undecane comprised more than 90% of all components and in a single Dufour's gland amounted to 19 microg. n-Decane and n-pentadecane were also included in the Dufour's gland secretion. Only formic acid was detected in the poison gland, in amounts ranging from 0.049 to 0.91 microl. This ant species releases a mixture of these substances, each of which has a different volatility and function. When the ants sensed formic acid, they eluded the source of the odor; however, they aggressively approached odors of n-undecane and n-decane, which are highly volatile. In contrast, n-pentadecane, which has the lowest volatility among the identified compounds, was shown to calm the ants. The volatilities of the alarm pheromone components were closely related to their roles in alarm communication. Highly volatile components vaporized rapidly and spread widely, and induced drastic reactions among the ants. As these components became diluted, the less volatile components calmed the excited ants. How the worker ants utilize this alarm communication system for efficient deployment of their nestmates in colony defense is also discussed herein.     

Revisiting the trail pheromone components of the red imported fire ant,Solenopsis invicta Buren

Ants use species-specific trail pheromones to coordinate their sophisticated foraging behavior. During the past decades, many trail pheromone components with various structures have been identified in ants, including the red imported fire ant, Solenopsis invicta, a notorious invasive species worldwide. Four compounds, Z,E- (ZEF) and E,E-α-farnesene (EEF), Z,E- (ZEHF) and E,E-α-homofarnesene (EEHF), have been reported as components of S. invicta trail pheromone. However, another study reported an analog of α-farnesene, Z,Z,Z-allofarnesene, as a key trail pheromone component. These contrasting results caused some uncertainty about the trail pheromone composition in S. invicta. In this study, we synthesized ZEF and EEF, ZEHF and EEHF, and reanalyzed the chemicals in the Dufour gland extract and in the trail pheromone fraction of S. invicta worker body extract. The reported isomers of farnesene and homofarnesene were detected and showed trail-following activity, with ZEF as the major compound, while no allofarnesene was found, neither in the Dufour gland extract nor in the whole-body extract. Our results confirm ZEF and EEF, ZEHF and EEHF as trail pheromone components of S. invicta.     

New chemicals from the dufour gland of the formicine ant Lasius niger (Hymenoptera:Formicidae)

A series of esters of C₈-C₁₃ fatty acids formed with C₁₀-C₁₂ straight chain alcohols, representing 3.7% of the glandular contents, was found in the Dufour gland of workers of the formicine ant Lasius niger. Altogether, nearly 50 components, among them hydrocarbons, alcohols, acetates and propionates were identified in the glandular liquid by means of gas chromatography (GC) and mass spectrometry (MS), using a solid injection technique. The wax esters and the propionates—decyl, dodecyl, octadecyl and farnesyl—have not been previously reported from ant secretions.     

膜翅目昆虫杜氏腺形态结构、内分泌物与功能的研究进展

杜氏腺为膜翅目雌性昆虫生殖器官中的一外分泌腺体,外形呈囊腔状,内含丰富的油状分泌物。其内分泌物主要为饱和与不饱和烃类化合物,有着辅助产卵、释放信息素及作幼虫食物等多方面的功能。本文对杜氏腺的形态结构、内分泌物合成途径、化学组分及其生理和生态学功能进行了综述。