Foliar and cortex oleoresin variability of silver fir (Abies alba Mill.) in Albania

Terpene composition of needle and cortical oleoresin from lateral shoots were analyzed by GC/MS for four Silver fir (Abies alba Mill.) populations scattered in natural species range in Albania. More than sixty compounds were detected in the needle oleoresin, which was characterized by a high content of alpha-pinene, camphene, beta-pinene, limonene and bornyl acetate. Three monoterpenes, alpha-pinene, beta-pinene and limonene, and two sesquiterpenes, beta-caryophyllene and germacrene D, comprised the majority of cortical oleoresin. The terpene composition differences among the populations that led to the recognition of two chemotypes. The needle oleoresin from the provinces of Puka, Bulqiza and Llogara were characterised by high amounts of beta-pinene, camphene and alpha-pinene and low amounts of limonene, while that from Drenova had high amounts of beta-pinene and limonene. A similar pattern was found in the cortical oleoresin with the exception of camphene that was a minor contributor. Geographical and seasonal variation between the populations was, also, investigated. Multivariate analysis of both needle and cortical oleoresin separated Drenova (southeastern population) from the other sites. When both major monoterpenes and sesquiterpenes were considered four chemical profiles could be attributed. Based on their chemical profiles, the populations can be divided into two groups: Populations with high content of beta-pinene and alpha-pinene but a low content of limonene (Puka, Bulqiza and Llogara), typical of most of A. alba populations in all its distribution range. Population with a high content of limonene and a moderate content of beta-pinene and alpha-pinene (Drenova).     

Odour-genes eovariance and differential investigation of individual odours in the Mus species complex

Previous research using habituation techniques has demonstrated that greater genetic similarity between two individuals is associated with more similarity in the qualities of their individual odours ('odour-genes eovariance'). We assessed odour similarities across species in two pairs of genetically close species within the Mus species complex (M. musculus and M. domesticus; M. spicilegus and M. macedonicus). Subjects treated odours within each species pair as similar compared with an odour from the other species pair. Subjects also treated odours of M. spicilegus males from the same population as similar compared with the odour of M. spicilegus males from a different population. This confirms odour-genes eovariance across species and within populations and also supports previous findings that odour similarities are more salient than specific odour markers. When adult males were presented with odours of females from two different heterospecific species, subjects spent more time investigating the odour from his own species pair than the other species pair, indicating greater interest in the odour of the closer heterospecific and demonstrating that odour-genes eovariance is reflected in behavioural responses to odours. Implications of odour-genes eovariance as a basis for identifying degrees of genetic relatedness of unfamiliar individuals through similarities in individual odours are discussed.     

PERMANENT GENETIC RESOURCES: Polymorphic microsatellite loci and interspecific cross-amplification in the parasitoid wasps Megastigmus stigmatizans and Megastigmus dorsalis

We isolated and characterized 19 polymorphic microsatellite loci in the congeneric parasitoid wasps Megastigmus stigmatizans and Megastigmus dorsalis associated with cynipid oak galls. Loci isolated from species-specific libraries showed extensive cross-amplification, resulting in a total of 15 polymorphic loci for M. stigmatizans and 13 for M. dorsalis.     

Electroantennogram responses of Douglas-fir seed chalcids to plant volatiles

Douglas fir seed chalcids females oviposit specifically into Douglas cones which they locate using olfactory and visual cues. We have tested whether this specialization was correlated with a specialized sensitivity towards cone volatiles. Field collected adults were presented a series of pure volatile chemicals. Electroantennogram responses (EAG) were recorded to generally occurring terpenoids and straight chain alcohols and aldehydes found in flowers and plant leaves. Monoterpenes identified in Douglas cone headspaces (alpha-pinene, beta-pinene, 3-thujene, alpha- and beta-phellandrene, gamma-terpinene and myrcene) elicited lower EAG responses than fatty acid derivatives corresponding to green odours. The EAG response profile of females differed significantly from that of males. Females were very sensitive to hexanol-1, heptanal, (Z)-3 hexenol-1, terpineol and terpinen-4-ol. Male responses were significantly higher to humid air, nerolidol, thujan-4-ol, hexanal, carvacrol, piperitone and farnesol. Several compounds (Z- verbenol, carvone, jasminol, geraniol, nerol and eugenol) elicited long lasting electrophysiological responses (over 5 s) in both sexes.     

Effects of in utero odorant exposure on neuroanatomical development of the olfactory bulb and odour preferences

Human babies and other young mammals prefer food odours and flavours of their mother's diet during pregnancy as well as their mother's individually distinctive odour. Newborn mice also prefer the individual odours of more closely related--even unfamiliar--lactating females. If exposure to in utero odorants-which include metabolites from the mother's diet and the foetus's genetically determined individual odour-helps shape the neuroanatomical development of the olfactory bulb, this could influence the perception of such biologically important odours that are preferred after birth. We exposed gene-targeted mice during gestation and nursing to odorants that activate GFP-tagged olfactory receptors (ORs) and then measured the effects on the size of tagged glomeruli in the olfactory bulb where axons from olfactory sensory neurons (OSNs) coalesce by OR type. We found significantly larger tagged glomeruli in mice exposed to these activating odorants in amniotic fluid, and later in mother's milk, as well as significant preferences for the activating odour. Larger glomeruli comprising OSNs that respond to consistently encountered odorants should enhance detection and discrimination of these subsequently preferred odours, which in nature would facilitate selection of palatable foods and kin recognition, through similarities in individual odours of relatives.     

Behavioural development of conspecific odour preferences in bank voles,Clethrionomys glareolus

Biological odours of conspecifics are known to have strong influences on behavioural interaction in bank voles Clethrionomys glareolus. This experiment tested two hypotheses. (1) Olfactory cues from familiar and unfamiliar mature opposite-sex conspecifics differ in their attractiveness to males and females, and their behavioural reactions change with age. (2) A genetically based mechanism is involved in female recognition of kin.In a two-choice preference test, prepubertal males and females were more attracted to familiar than to unfamiliar odours of opposite-sex conspecifics, as manifested by more time spent sniffing familiar voles. As the young reached sexual maturity they shifted their odour preferences. Mature males and females preferred the novel odour of unrelated opposite-sex conspecifics to that of relatives. The results of experiments testing the second hypothesis indicate that females use a genetically based mechanism to recognise their kin. Young and mature females were able to recognise the odour of their biological but socially unknown fathers, and showed the same pattern of behaviour as females in previous experiments.The possible biological functions of kin recognition in bank voles are discussed.     

Larval and adult nutrition effects on blood/nectar choice of Culex nigripalpus mosquitoes

Abstract. The impact of nutritional variables on the development of host-seeking and biting behaviours after emergence by female Culex nigripalpus mosquitoes were studied using air-flow olfactometer and close-range biting assays, respectively. Unfed females failed to develop resting stage ovarian follicles. When offered a bird host in the absence of competing stimuli, sugar-fed mosquitoes were significantly more responsive in both host-seeking and biting than unfed controls. In a choice olfactometer assay using nectar odours (honey scented with artificial apple-blossom oil) versus host odours (a bird), unfed females preferred honey over bird odours except when honey odour was weak. After sucrose feeding, females switched from honey to bird preference. This change in behaviour was accompanied by significant accumulation of lipid and by follicular growth to the resting stage. Elevation of host responsiveness after sugar feeding was reversible; starvation ultimately resulted in females preferring honey over bird odours. When the larval diet was restricted by crowding, the wing-length and total lipid of resultant adult females were reduced. Although differences were subtle, unfed bird-responding females tended to have longer wings and more lipid than their honey-responding counterparts.     

MHC-correlated odour preferences in humans and the use of oral contraceptives

Previous studies in animals and humans show that genes in the major histocompatibility complex (MHC) influence individual odours and that females often prefer odour of MHC-dissimilar males, perhaps to increase offspring heterozygosity or reduce inbreeding. Women using oral hormonal contraceptives have been reported to have the opposite preference, raising the possibility that oral contraceptives alter female preference towards MHC similarity, with possible fertility costs. Here we test directly whether contraceptive pill use alters odour preferences using a longitudinal design in which women were tested before and after initiating pill use; a control group of non-users were tested with a comparable interval between test sessions. In contrast to some previous studies, there was no significant difference in ratings between odours of MHC-dissimilar and MHC-similar men among women during the follicular cycle phase. However, single women preferred odours of MHC-similar men, while women in relationships preferred odours of MHC-dissimilar men, a result consistent with studies in other species, suggesting that paired females may seek to improve offspring quality through extra-pair partnerships. Across tests, we found a significant preference shift towards MHC similarity associated with pill use, which was not evident in the control group. If odour plays a role in human mate choice, our results suggest that contraceptive pill use could disrupt disassortative mate preferences.     

Role of visual cues and interaction with host odour during the host-finding behaviour of the cabbage moth

The approach and landing responses of female Mamestra brassicae (L.) (Lepidoptera: Noctuidae) to visual cues from artificial plant leaves of different shapes and presence/absence of cabbage plant odour were investigated in a laboratory wind tunnel. The leaves were painted with cadmium yellow colour and observed under dim red light. Females showed oriented flight towards plant odours but landed significantly more often when the odour was presented with an artificial leaf. In three-choice tests, the shape of the leaf targets (circle, square or triangle) did not influence the female response. However, the size of the target did influence the insect response: the females preferred landing on square targets with sides of 5 or 10 cm rather than on the largest target, with sides of 15 cm. The orientation of the target influenced the insects' response: females landed significantly more often on the target positioned vertically than horizontally.     

Variability in odour reception in the peripheral sensory system of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

Abstract  Mixtures of single odours were used to explore the receptor response profile across individual antennae of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). Seven odours were tested including floral and green-leaf volatiles: phenyl acetaldehyde, benzaldehyde, β-caryophyllene, limonene, α-pinene, 1-hexanol, 3 Z -hexenyl acetate. Electroantennograms of responses to paired mixtures of odours showed that there was considerable variation in receptor tuning across the receptor field between individuals. Data from some moth antennae showed no additivity, which indicated a restricted receptor profile. Results from other moth antennae to the same odour mixtures showed a range of partial additivity. This indicated that a wider array of receptor types was present in these moths, with a greater percentage of the receptors tuned exclusively to each odour. Peripheral receptor fields show variation in the spectrum of response within a population (of moths) when exposed to high doses of plant volatiles. This may be related to recorded variation in host choice within moth populations as reported by other authors.     

濒危植物秦岭冷杉结实特性的研究

秦岭冷杉 (Abieschensiensis) 是松科冷杉属常绿乔木, 为我国特有珍稀濒危植物。该文在秦岭冷杉分布最为集中的秦岭地区分高、低 2个海拔高度采集当年球果和种子, 通过野外观察、室内实验和对比分析, 研究秦岭冷杉的结实特征以及球果和种子的基本性状, 并探讨这些特征在不同海拔高度的差异。结果表明, 与冷杉属其它树种相比, 秦岭冷杉球果和种子较大、单果种子数较多, 但空粒多、饱满度低。自然状况下饱满度为 12.0 %, 人工目视分选后饱满度 2 1.6 %, 风选后可达到 38.1%, 其余种子大部分为空粒、涩粒, 有少量虫粒。随海拔高度增加, 秦岭冷杉天然群体成龄个体中结实母树比例降低, 结实量减少。秦岭冷杉总体结实母树比例为 5 5 %, 低海拔处结实母树比例较高, 为 76 %, 高海拔结实母树比例为 34%。秦岭冷杉单株母树的平均结实量为 113个, 结实量小于 10 0个的母树占 6 4.5 %, 其中 5 0 %的母树结实量在 5 0~ 10 0个, 结实量在 10 0~ 2 0 0个的占 16.1%, 2 0 0个以上的占 19.4 %。 2 0 0个以上球果的个体均生长在低海拔区间, 高海拔处结实量多在 10 0个以下。方差分析表明, 两个海拔高度间球果和种子特征差异是显著的, 绝大部分球果和种子性状指标的平均值在低海拔处大于高海拔处。因此, 从结实特性以及球果和种子的特征来看, 低海拔处秦岭冷杉生殖生长好于高海拔处。     

Individual odours and mate recognition in the prairie vole,Microtus ochrogaster

A habituation-discrimination technique was used to demonstrate that male and female prairie voles can discriminate individual differences in the odours of soiled shavings and urine from male and female conspecifics. A second experiment, employing a Y-maze, showed that females significantly preferred the odours of their mate over those of either another mated male or an unmated male. Males preferred their mate's odours to those of other mated females but showed no significant preference between the odours of their mate and those of a virgin female. A third experiment demonstrated that, over a 10-h period, females built nests and/or stayed preferentially on the side of a Y-maze containing their mate's odours. Likewise, males preferentially built nests in their mate's side compared to the side containing odours from a virgin female. However, although the same trend was present when mate odours were paired with odours from another mated female, the preference was not statistically significant. Taken as a whole, these results indicate that mate recognition may be an important of individually distinctive odours in this species.     

A six-arm olfactometer permitting simultaneous observation of insect attraction and odour trapping

Abstract.  Behavioural assays to study insect attraction to specific odours are tedious, time consuming and often require large numbers of replications. Olfactometer and flight tunnel tests can usually only be conducted with one or two odour sources at a time. Moreover, chemical information on the odour sources has to be obtained in separate analytical studies. An olfactometer was developed in which six odours can be tested simultaneously for their relative attractiveness while during the assays, part of each test odour can be trapped for further analyses. The effectiveness of this six-arm olfactometer was tested by observing the responses of the solitary endoparasitoid Cotesia marginiventris (Cresson) to host-induced odours from young maize plants. For statistical analyses, we used log-linear models were adapted to account for overdispersion and possible positional biases. Female wasps responded extremely well in tests where they were offered a single odour source, as well as in tests with multiple choices. The responses of wasps released in groups were the same as those released individually and it was found that females did not attract or repel each other, but males preferred arms in which females had been released. Dose–response tests with varying numbers of plants or host larvae on plants revealed that the wasps responded in a dose-related manner, thus showing that the system is well suited to measure relative preference. The clear choices of the insects amongst six possibilities provided substantial statistical power. Gas chromatographic analyses of sampled air revealed clean and effective odour trapping, which largely facilitates the comparison of results from behavioural assays with the actual blends of volatiles that were emitted by the various odour sources. Advantages and disadvantages compared to other methods are discussed.     

Chemical composition and antimicrobial activity of essential oil from cones of Pinus koraiensis

The essential oil from the cones of Pinus koraiensis was prepared after removing the seeds, and its chemical composition analyzed using gas chromatography-mass spectrometry (GC-MS). Hydrodistillation of the P. koraiensis cones yielded 1.07% (v/w) of essential oil, which was almost three times the amount of essential oil extracted from the needles of the same plant. Moreover, the antimicrobial activities of the oil against the growth of Gram-positive bacteria, Gram-negative bacteria, and fungi were evaluated using the agar disc diffusion method and broth microdilution method. Eighty-seven components, comprising about 96.8% of the total oil, were identified. The most abundant oil components were limonene (27.90%), alpha-pinene (23.89%), beta-pinene (12.02%), 3-carene (4.95%), beta-myrcene (4.53%), isolongifolene (3.35%), (-)-bornyl acetate (2.02%), caryophyllene (1.71%), and camphene (1.54%). The essential oil was confirmed to have significant antimicrobial activities, especially against pathogenic fungal strains such as Candida glabrata YFCC 062 and Cryptococcus neoformans B 42419. Therefore, the present results indicate that the essential oil from the cones of Pinus koraiensis can be used in various ways as a nontoxic and environmentally friendly disinfectant.     

Pheromonal Recognition of Females Takes Precedence over the Chromatic Cue in Male Iberian Wall Lizards Podarcis hispanica

Sex recognition is based on colour signals in many species of lizards. However, olfactory stimuli are also clearly involved, and many species might rely more on chemoreception. We aimed to examine whether colour pattern or odours, or both, are used in sex recognition and which cues elicit courtship of females by males of the lizard Podarcis hispanica . We experimentally manipulated the coloration and odour of female P. hispanica , thereby creating groups with all combinations between coloration and odour of males and females. Using data from staged encounters, we compared the responses of resident males to manipulated and unmanipulated individuals (males and females). Responding males reacted significantly more aggressively to female intruders with male odours, independently of their coloration. Nevertheless, coloration seemed to be important in long-distance sex recognition since, in the first minutes, females painted as females received a lower number of aggressive responses. Both colour and odour were important in eliciting male courtship. However, females painted as females and with female odours were preferentially courted. Comparisons with unmanipulated male and female intruders agreed with these expectations. Therefore, at close range, odoriferous cues seem to be more important than colour patterns in sex recognition, but female coloration is also useful at long range to deter the aggressive response of males and to elicit courtship in conjunction with odours.     

幼小鼠对种内几种化学信息的识别和反应

50年代以来,国际上相继开展了一些关于化学信息在小家鼠繁殖中作用的研究,并在实验室中揭示了一些反应:如集群雌鼠气味使雌鼠性周期延长的Lee-Boot效应,雌鼠气味能解除集群雌鼠对性周期的抑制、并促使它们同期发情的Whitten效应,以及陌生雄鼠气味中断妊娠的Bruce效应等等。但是,在成鼠间起作用的几种化学信息,是否能被幼鼠感受,对幼鼠的发育和性成熟是否产生影响,还是一个未知的问题。 影响小鼠发育的因素很多,除环境因子外,近来还出现了有关社群因素中外激素对小鼠性成熟影响的报道,发现雄鼠或给予雄激素的雌鼠有促进幼雌鼠性成熟的作用,范志勤发现父本气味和陌生雄鼠气味对幼雌鼠性成熟均有促进作用。但是,目前关于种内雌鼠气味、集群雌鼠气味、幼鼠气味影响性成熟的研究尚不多见。有关各种气味对性成熟效应的比较的报道亦少。本文的目的在于通过实验方法,阐明种内的几种化学信息,诸如父本、陌生雄性、雌性、集群雌性、幼雌等几种气味,对幼雌小鼠性成熟的影响,并据此了解幼鼠识别这几种信息的能力。     

Chemical Composition of the Essential Oils from Different Morphological Parts of Pinus cembra L.

The essential oils from needles, twigs, bark, wood, and cones of Pinus cembra were analyzed by GC‐FID, GC/MS, and ¹H‐NMR spectroscopy. More than 130 compounds were identified. The oils differed in the quantitative composition. The principal components of the oil from twigs with needles were α‐pinene (36.3%), limonene (22.7%) and β‐phellandrene (12.0%). The needle oil was dominated by α‐pinene (48.4%), whereas in the oil from bark and in the oil from twigs without needles there were limonene (36.2% and 33.6%, resp.) and β‐phellandrene (18.8% and 17.1%, resp.). The main constituents of the wood oil as well as cone oil were α‐pinene (35.2% and 39.0%, resp.) and β‐pinene (10.4% and 18.9%, resp.). The wood oil and the cone oil contained large amounts of oxygenated diterpenes in comparison with needle, twig, and bark oils.     

Oviposition Sites of the Cypress Seed Bug <Emphasis Type="Italic">Orsillus maculatus</Emphasis> and Response of the Egg Parasitoid <Emphasis Type="Italic">Telenomus</Emphasis> gr. <Emphasis Type="Italic">Floridanus</Emphasis>

Phytophagous insects have many strategies to escape parasitoids, for example by hiding eggs into plant tissues, but oviposition in holes made by another insect is rather scarce. The cypress seed bug Orsillus maculatus Fieber (Heteroptera: Lygaeidae) is strictly dependent on the availability of cones of Cupressus sempervirens L. to oviposit. Females lay eggs either in exit holes cut through the cone scale by emerging adults of the cypress seed chalcid, Megastigmus wachtli Seitner (Hymenoptera: Torymidae), or on the inner side of scales of partly open cones. A recently discovered egg parasitoid belonging to the genus Telenomus has been shown to attack bug eggs in both oviposition sites. In this paper we investigated the parasitoid performance according to oviposition sites. Field samplings were conducted in two evergreen cypress orchards located in the south of France. The distribution and condition of the egg patches were compared between the two locations and oviposition sites. Seed bugs preferred to oviposit in emergence holes of M. wachtli, and parasitoid performance was higher in eggs laid on cone scales. The chalcid emergence holes seemed to ensure bug eggs with enemy-free space. Oviposition site selection could be an adaptive strategy to escape parasitoid attack.     

Scent-marking behaviour in response to conspecific odours by the rat, Rattus norvegicus

The scent-marking behaviour of male and female rats in response to conspecific odours was investigated in two experiments. The results of the first experiment indicated that exposure to conspecific odours generally led to an increase in marking rates. However, no sex differences were found, and large differences in marking responses according to the sex of the odour donor were only found for dioestrous females. These results do not suggest that the primary function of scent-marking in this species is sexual. In the second experiment, odour sources differed not only in the sex of the donor, but also in their relative familiarity to the animal being tested. This experiment showed that males marked more in response to odours from familiar (non-related) females. Females did not differ significantly on measures of marking but did display greater interest in odours from unfamiliar males. The results are discussed in relation to possible functions of marking in this species.     

Cross-Fostering of Male Mice Subtly Affects Female Olfactory Preferences

The maternal environment has been shown to influence female olfactory preferences through early chemosensory experience. However, little is known about the influence of the maternal environment on chemosignals. In this study, we used two inbred mouse strains, C57BL/6 (C57) and BALB/c (BALB), and explored whether adoption could alter male chemosignals and thus influence female olfactory preferences. In Experiment 1, C57 pups were placed with BALB dams. Adult BALB females then served as the subjects in binary choice tests between paired male urine odours (BALB vs. C57, BALB vs. adopted C57 and C57 vs. adopted C57). In Experiment 2, BALB pups were placed with C57 dams, and C57 females served as the subjects in binary choice tests between paired male urine odours (C57 vs. BALB, C57 vs. adopted BALB, and BALB vs. adopted BALB). In both experiments, we found that females preferred the urine of males from different genetic backgrounds, suggesting that female olfactory preferences may be driven by genetic compatibility. Cross-fostering had subtle effects on female olfactory preferences. Although the females showed no preference between the urine odours of adopted and non-adopted males of the other strain, the BALB females preferred the urine odour of BALB males to that of adopted C57 males, whereas the C57 females showed no preference between the urine odour of C57 and adopted BALB males. Using gas chromatography-mass spectrometry (GC-MS) and stepwise discriminant analysis, we found that the ratios of volatile chemicals from urine and preputial gland secretions were altered in the fostered male mice; these changes may have resulted in the behavioural changes observed in the females. Overall, the results suggest that female mice prefer urine odours from males with different genetic backgrounds; this preference may be driven by genetic compatibility. The early maternal environment influences the chemosignals of males and thus may influence the olfactory preferences of females. Our study provides additional evidence in support of genotype-dependent maternal influences on phenotypic variability in adulthood.