Effect of gland extracts on digging and residing preferences of red imported fire ant workers (Hymenoptera: Formicidae)

There is evidence that ant‐derived chemical stimuli are involved in regulating the digging behavior in Solenopsis invicta Buren. However, the source gland(s) and chemistry of such stimuli have never been revealed. In this study, extracts of mandibular, Dufour's, postpharyngeal, and poison glands were evaluated for their effect on ant digging and residing preferences of S. invicta workers from three colonies. In the intracolonial bioassays, workers showed significant digging preferences to mandibular gland extracts in 2 of 3 colonies and significant residing preferences in 1 of 3 colonies; significant digging preferences to Dufour's gland extracts in 1 of 3 colonies and significant residing preferences in 2 of 3 colonies. No digging and residing preferences were found for postpharyngeal and poison gland extracts. In intercolonial bioassays, significant digging and residing preferences were found for mandibular gland extracts in 3 of 6 colony combinations. Significant digging preferences to Dufour's gland extracts were found in 4 of 6 colony combinations and significant residing preferences in all 6 colony combinations. For postpharyngeal gland extracts, significant digging preferences were found only in 1 of 6 colonial combinations and no significant residing preferences were found. For poison gland extracts, no significant digging preferences were found; significant residing preferences were found in 1 of 6 colony combinations. However, a significant residing deterrence (negative residing preference index) was found for 2 of 6 colony combinations. Statistical analyses using data pooled from all colonies showed that mandibular and Dufour's gland extracts caused significant digging and residing preferences in both intracolonial and intercolonial bioassays but not postpharyngeal and poison gland extracts. By analyzing the data pooled from the same three colonies used for gland extract bioassays, it was found that, in no cases, workers showed significant digging and residing preferences to 2‐ethyl‐3,6‐dimethylpyrazine, an alarm pheromone component from mandibular gland.     

Caste and colony variations in the chemical composition of the cephalic secretions of the African weaver ant, Oecophylla longinoda

ABSTRACT. Considerable differences were found in the composition of the volatile cephalic chemical of the castes of Oecophylla longinoda. The mandibular glands of minor workers contain a series of primary alcohols, predominantly 1-hexanol, 1-octanol and 1-nonanol, together with nerol and geraniol. Hexanal, 2-butyl-2-octenal and 3-undecanone (components of the mandibular gland secretion of major workers, and which are important stimuli in alarm communication) are absent. Major workers are attracted and arrested by high concentrations of nerol, which is part of a possible mechanism for communication within the nest. The heads of males contain a pungent secretion, which may be used in defence; the major constituents are five aliphatic acids. The quantities of both main and trace constituents of the mandibular gland secretion of major workers were found to vary between colonies collected in different areas of West Africa.     

Chemical signals of queens in kin recognition of honeybees,Apis mellifera L.

Summary The volatile signals of live queens as well as head and tergite extracts were analyzed by capillary gas chromatography. Multivariate analysis revealed a significant decrease in the variability of the volatile signals of queens the more closely related they were. Though extracts of the tergal glands showed the same phenomenon, head extracts of closely related queens had no significantly reduced variabilities compared to extracts of unrelated queens. Workers, tested in a metabolic bioassay, could not discriminate among head extracts of related and unrelated queens. Therefore the classical queen substance and the semiochemicals of the mandibular glands are unlikely to be used as kin recognition labels by the workers. Workers could, however, discriminate among the odours of tergite extracts and the volatile signals of queens, indicating that the signal of the tergal gland secretions is a possible source for kin recognition labels which the mandibular gland secretions are not. Furthermore the results show that learning is strongly involved in the kin recognition system of honeybees.     

Female mating receptivity after injection of male-derived extracts in Callosobruchus maculatus

The effects of male-derived extracts on female receptivity were investigated in Callosobruchus maculatus (Coleoptera: Bruchidae). Injection of aqueous extracts of the male reproductive tract into the abdomen of females reduced receptivity. Aqueous extracts of male reproductive tracts were divided to three molecular weight (MW) fractions by ultrafiltration: Fractions: (I) MW<3 kDa, (II) 3-14 kDa, and (III)>14 kDa. Fraction II reduced female receptivity from 3h after injection, and Fraction III reduced female receptivity from 2 days after injection. On the other hand, no effect on receptivity was found for Fraction I. Furthermore, male reproductive tract organs were divided into accessory gland, testis, and seminal vesicle including the ejaculatory duct. Aqueous extracts of the seminal vesicle reduced receptivity of females immediately following injection, while aqueous extracts of the accessory gland reduced receptivity at the second day. The results suggest that the components of Fraction II existed in the seminal vesicle, and those of Fraction III in the accessory gland. The results of the present and the previous studies in Callosobruchus chinensis, a species closely related to C. maculatus, were compared and are discussed from the viewpoint of the significance of ejaculation in the two species.     

Putative chemosignals of the ferret (Mustela furo) associated with individual and gender recognition

Quantitative stir bar sorptive extraction methods, both in the aqueous and headspace modes, followed by thermal desorption gas chromatography-mass spectrometry were used to investigate individual variations in the volatile components of male and female ferret (Mustela furo) urine. The urinary profiles were further compared with volatile profiles of anal gland secretions of breeding male and female ferrets. Thirty volatile compounds were quantified in male and female urine. Among them, 2-methylquinoline was unique to male urine. Four ketones (4-heptanone, 2-heptanone, o-aminoacetophenone, and a dimethoxyacetophenone) and several nitrogen compounds (e.g., 2,5-dimethylpyrazine, quinoline, 4-methylquinazoline) and low levels of three unidentified nonsulfur compounds were significantly more abundant in males than in females. Quantitative comparison of 30 volatile urinary compounds showed several statistically significant differences between the sexes and individuals of the same sex. These findings suggest that ferrets may use urine marking for sex and individual recognitions. Ten of the 26 compounds identified in anal gland secretions from females and males were also found in urine. However, most of the major compounds (thietanes, dithiolanes, and indole) in anal glands were not present in urine. This suggests that urine may convey specific signals that differ from those of anal glands. Additionally, 10 volatiles (two aldehydes, five ketones, benzothiazole, 2-methylquinoline, and 4-methylquinazoline), not previously identified, were found in ferret anal gland secretions. Among the new compounds, o-aminoacetophenone was found only in males, while only traces of this compound were found in females. Similar results were previously obtained in anal glands of three other Mustela species. These findings provide new information about the constituents of urine and volatile components of anal gland secretions in ferrets.     

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.     

Volatile components from the mandibular glands of the turtle ants, Cephalotes alfaroi and Cephalotes cristatus

GC–MS analysis of whole head extracts from the turtle ants, Cephalotes alfaroi and Cephalotes cristatus, showed that 4-heptanone and 4-heptanol were the major volatile components in the mandibular glands. 4-Heptanone and 4-heptanol have rarely been identified in mandibular gland secretions from other ant genera. Thus, these compounds may be chemotaxonomic markers for the genus Cephalotes, since they have been identified in the mandibular glands from all members of this genus that have been investigated to date. Minor components identified in the whole head extracts of these ants were 3-methyl-1-butanol, 3-heptanone, 3-hexanol, 2- and 3-methylbutanoic acids, 2-methyl-4-heptanone, 2-phenylethanol and phenol. To our knowledge, this is the first time that 2-methyl-4-heptanone and phenol have been reported in the mandibular gland secretion from any Formicid.     

The ecological and taxonomic importance of flower volatiles of Clusia species (Guttiferae)

The chemical composition of floral volatiles of sixteen species of Clusia (Guttiferae) belonging to four different taxonomic sections of the genus was investigated. The volatiles were extracted from fresh petals by microhydrodistillation and analysed by GC/MS. The composition of the volatiles was in part, but not always, related to the taxonomic position of the species, and to a minor extent to the type of pollinator observed on the flowers as revealed by clustering analysis. The composition of the volatile components of female and male flowers belonging to the same species (C. grandiflora, C. lanceolata, C. paralicola, C. parviflora and C. spiritu-sanctensis) was found to be almost identical. Field bioassays showed the petal extracts to be attractive to pollinating bees.     

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.     

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。     

The mandibular gland secretion of the ant, Myrmica scabrinodis

ABSTRACT. The volatile secretion of the mandibular gland of the common elbowed red ant, Myrmica scabrinodis Nyl., is shown to consist of ethanal, ethanol, acetone, 3-hexanone, 3-hexanol, 3-heptanone, 3-heptanol, 3-octanone, 3-octanol, 6-methyl-3-octanone, 6-methyl-3-octanol, 3-nonanone, 3-nonanol, 3-decanone and 3-undecanone. The electroantennographic response to the major components was recorded and compared with some related compounds. Behavioural tests were carried out on the major constituents, showing that 3-octanol is an attractant for workers, that 3-octanone increases the effect of 3-octanol, and 3-nonanone augments the linear speed of the ants.     

Alarm pheromones of the Attini: Their phylogenetic significance

The volatile components present in the mandibular glands of a number of species of the attine genera Cyphomyrmex (1 species), Trachymyrmex (3 species), and Acromyrmex (2 species) were investigated and compared with those present in Atta. The extracts were found to consist of mixtures of a number of compounds. All but one of these mixtures contained some or all of the following compounds: 3-octanone, 4-methyl-3-heptanone, 3-octanol, and 4-methyl-3-heptanol. The behavioural responses of Trachymyrmex and Cyphomyrmex workers to these compounds were tested. A common chemical heritage based on 3-ketones and 3-alcohols appeared to exist among the genera studied. The chemical data were compared with an accepted phylogeny of these genera to see whether it supported the phylogeny.     

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.     

Metasternal gland secretion of the locust tree borer, Megacyllene robiniae (Coleoptera: Cerambycidae)

1. The volatile components of metasternal gland extracts of male and female Megacyllene robiniae have been analyzed by gas chromatography-mass spectroscopy. 2. The major component was identified as 2-(1,3-hexadien-1-yl)-5-methyltetrahydrofuran, a new natural product. 3. 1-Phenylethanol is present only in male extracts. 4. Acetates of hexadecanol and octadecanol are also present.     

The ontogenetic pattern of mandibular gland components in queenless worker bees (Apis mellifera capensis Esch.)

The quantity and composition of the six major mandibular gland components of young queenless workers of the Cape honeybee (Apis mellifera capensis) were determined. The total amount of the six components increased with age. The relative quantities in the mandibular gland secretion of queenless caged workers were found to change rapidly during the first 4 days after emergence and to become dominated by the queen substance, 9-keto-2(E)-decenoic acid. Also the relative amounts of 9-hydroxy-decenoic acid, a precursor of the queen substance, showed an increase of an order of magnitude within the first 4 days of imaginal life. The relative amounts of the aromatic compounds typical to the queen pheromone remained similar in this developmental time window. The increase of queenlike compounds is particularly strong between days two and three after emergence. These queen-like pheromones play a major role in the development of reproductive hierarchies among workers under queenless conditions. This may be an important factor in the socio-parasitic pathway of A. m. capensis.     

Alkylpyrazines in ponerine ants: Their presence in three genera,and caste specific behavioural responses to them in Odontomachus troglodytes

Alkylpyrazines were found in the mandibular gland secretions of three Ethiopian ponerine ants. The major components in Odontomachus troglodytes were 2,6-dimethyl-3-n-butyl- and n-pentylpyrazine; in Anochetus sedilloti 2,6-dimethyl-3-sec-butyl- and 2,5-dimethyl-3-n-pentylpyrazine, and in Brachyponera sennaarensis 2,6-dimethyl-3-n-butyl and n-pentylpyrazines. The male caste in O. troglodytes contained 2,6-dimethyl-3-n-hexyl- and n-butylpyrazine. In O. troglodytes caste-specific behavioural differences were found between males and workers in response to gland equivalent quantities of their pheromones. The fragile males retreated from the alkylpyrazines whereas the workers were attracted to and attacked the pheromone source. Large quantities (5 gland equivalents) of pheromones were also repellent to the workers.     

Hydrolases in the hypopharyngeal glands of workers of Scaptotrigona postica and Apis mellifera (Hymenoptera, Apinae)

Hydrolytic enzymes from hypopharyngeal gland extracts of newly emerged, nurse and foraging workers of two eusocial bees, Scaptotrigona postica, a native Brazilian stingless bee, and the Africanized honey bee (Apis mellifera) in Brazil, were compared. The hypopharyngeal gland is rich in enzymes in both species. Fifteen different enzymes were found in the extracts, with only a few quantitative differences between the species. Some of the enzymes present in the extracts may have intracellular functions, while others seem to be digestive enzymes. Scaptotrigona postica, had lower beta-glucosidase and higher lipase esterase activities than A. mellifera. The differences may be due to different feeding habits and behavioral peculiarities of the two species.     

Ontogenetic patterns of volatiles identified in Dufour's gland extracts from queens and workers of the primitively eusocial halictine bee,Lasioglossum malachmum (Hymenoptera: Halictidae)

Summary Ontogenetic patterns of volatile compounds identified in Dufour's gland extracts from queens and workers of the primitively eusocial sweat beeLasioglossum malachurum (K.) were compared. Only young unmated queens showed high proportions of isopentenyl esters, while macrocyclic lactones were dominant in old breeding queens, spring queens, and workers. In young queens the relative and absolute amounts of volatiles changed one day after mating. A discriminant analysis revealed significant differences in odor patterns of unmated and mated young queens. The fat body was the largest in young females, while eggs could be recorded only in breeding queens. Possible functions of different odor components in the investigated female groups are discussed.     

Secretions of stingless bees: cephalic secretions of two Frieseomelitta species

Gas chromatographic and mass spectrometric analysis of the volatile compounds in the mandibular glands of Frieseomelitta varia and the heads of Frieseomelitta silvestrii have revealed relatively simple or more complex mixtures of volatile oxygenated compounds. 2-Alkanols were found to be important components for both species. In F. varia the composition was essentially the same in samples from two widely separated areas and there were small differences between callows and mature, foraging bees. The first electroantennographic studies on stingless bees, using hexane extracts of heads of both species, have demonstrated a response in the workers’ antennae to these, as well as to the pure compounds 2-heptanol and 2-nonanol .     

Factors in the rat submaxillary gland that stimulate growth of cultured glioma cells: identification and partial characterization

The effect of rat submaxillary extract on the growth of rat C6 glioma cells in serum-free culture has been examined. Extracts (10-15 microgram/ml) of submaxillary glands from both male and female rats markedly enhanced the growth of serum-deprived C6 cells and, in combination with insulin, transferrin, and NIH-LH (a source of fibroblast growth factor), were able to stimulate C6 cell growth to an extent comparable to that achieved with an optimal amount of fetal calf serum. The mitogenic activity of rat submaxillary extracts was found to be heat-labile, acid-stable, and partially inactivated by protease and 2-mercaptoethanol. Under our assay conditions, biologically active preparations of purified mouse submaxillary gland epidermal growth factor (EGF) or nerve growth factor (NGF) were not mitogenic for C6 cells, nor was the mitogenic activity of rat submaxillary extracts inhibited by antiserum to these mouse submaxillary gland growth factors. These results suggest that the active component(s) of rat submaxillary extracts is unrelated to either EGF or NGF. The growth-enhancing effect also appears unrelated to esteropeptidase activity present in these extracts since the mitogenic activity was unaffected by several protease inhibitors. Moreover, two purified mouse submaxillary gland arginylesteropeptidases, EGF-binding protein and gamma-subunit of 7 S NGF, were unable to elicit a comparable growth response even when added to cell culture medium at unreasonably high concentrations. The C6 cell mitogenic activity of crude submaxillary extracts could be separated into two biologically similar components by either gel filtration on Sephadex G-100, preparative isoelectric focusing in a pH gradient of 3-10, or adsorption to DEAE-cellulose followed by elution with a sodium chloride gradient. One of the active components was acidic in nature and had an apparent molecular weight of 40,000, while the other was near neutral in charge and possessed a molecular weight of approximately 20,000. The relationship between these two C6 cell mitogenic components and the rat submaxillary gland component responsible for stimulating Balb/c-3T3 cell growth in serum-free, factor supplemented medium (McClure et al., 1979, J. Cell Biol. 83:96a) is also discussed.