“Pheromones and Animal Behaviour” - Communication by Smell and Taste”

Summary.  ((no summary.))A corrected version of the appendix is available for free download from     

Defensive chemistry of lycid beetles and of mimetic cerambycid beetles that feed on them

Summary.  Beetles of the family Lycidae have long been known to be chemically protected. We present evidence that North American species of the lycid genera Calopteron and Lycus are rejected by thrushes, wolf spiders, and orb-weaving spiders, and that they contain a systemic compound that could account, at least in part, for this unacceptability. This compound, a novel acetylenic acid that we named lycidic acid, proved actively deterrent in feeding tests with wolf spiders and coccinellid beetles. Species of Lycuscommonly figure as models of mimetic associations. Among their mimics are species of the cerambycid beetle genus Elytroleptus, remarkable because they prey upon the model lycids. We postulated that by doing so Elytroleptus might incorporate the lycidic acid from their prey for their own defense. However, judging from analytical data, the beetles practice no such sequestration, explaining why they remain relatively palatable (in tests with wolf spiders) even after having fed on lycids. Chemical analyses also showed the lycids to contain pyrazines, such as were already known from other Lycidae, potent odorants that could serve in an aposematic capacity to forestall predatory attacks. David Utterback: Deceased     

Differential responses in American (<Emphasis Type="Italic">Castanea dentata</Emphasis> Marshall) and Chinese (<Emphasis Type="Italic">C. mollissima</Emphasis> Blume) chestnut (Falales: Fagaceae) to foliar application of jasmonic acid

Summary.  Tannins are plant defense compounds that exhibit antibiotic (e.g. toxic) and antixenotic (e.g. repellent) effects against an array of plant pests. They are broadly divided into two major groups, hydrolysable tannins (gallotannins and ellagitannins) and proanthocyanidins, each with an undetermined number of compounds. We investigated constitutive levels of hydrolysable tannins and proanthocyanidins in the leaves and stems of American (Castanea dentata Marshall) (Fagales: Fagaceae) and Chinese (C. mollissima Blume) chestnut. American chestnut contained more proanthocyanidins in leaves and stems than Chinese chestnut, but Chinese chestnut contained more foliar hydrolysable tannins. Regardless of these differences, gypsy moth (Lymantria dispar L, Lepidoptera: Lymantriidae) performance did not differ when fed American and Chinese chestnuts. We also investigated the effects of jasmonic acid (JA) treatment on differential tannin induction in American and Chinese chestnut leaves and stems. JA treatment increased proanthocyanidins in American chestnut stems and hydrolysable tannins in both tissue types of American chestnut, but did not influence tannin concentrations in Chinese chestnut leaves or stems. HPLC-ESI-MS analysis of pooled samples suggested that hydrolysable tannins in each tissue were qualitatively comprised primarily of ellagitannins, and JA generally increased the number of hydrolysable tannins that could be detected by ESI-MS. Third, we investigated the performance of gypsy moths on JA treated and untreated American and Chinese chestnut. Caterpillar relative growth was not influenced by JA treatment on Chinese chestnut, but decreased in response to JA application on American chestnut. Our results indicate that JA-dependent defenses differ between these chestnut species. This study improves our understanding of ecologically important differences in tannin induction and herbivore susceptibility in Castanea, and has important implications in efforts for American chestnut restoration and commercial chestnut production.     

Abundance and diversity of the phyllosphere bacterial communities of Mediterranean perennial plants that differ in leaf chemistry

Summary.  We studied the epiphytic bacterial communities of the summer leaves of eight perennial species naturally occurring in a Mediterranean ecosystem. The species differ in essential-oil content (from rich in essential oil to non-producers) and composition, and also in life form (from herbaceous species to tall shrubs). We compared the epiphytic bacterial communities on the basis of (i) their abundance, (ii) their metabolic profile (derived by use of the BIOLOG Ecoplate system) and (iii) richness and diversity of substrates that they use, as a measure of functional diversity. Among all species, the aromatic Melissa officinalis was the most abundantly colonized. The bacterial communities on the leaves of the aromatic Myrtus communis, Calamintha nepeta and Melissa officinalis, and also of Cistus incanus catabolized all 31 substrates offered; those on the evergreen-sclerophyllous species, Arbutus unedo and Quercus coccifera, catabolized only 14 and 17 substrates, respectively. Carbohydrates were consistently used abundantly by all communities, whereas carboxylic acids were most variably used. On average, the group of aromatic plants scored higher regarding bacterial abundance, and richness and diversity of substrates used by the bacterial communities on their leaves; the lowest values for both substrate-use indices were recorded in A. unedo. Bacterial abundance or richness or diversity of substrates used did not vary with leaf oil content. Abundance was positively correlated with both substrate-use indices. Results support claims that the antimicrobial effects of essential oils are not exerted so much under natural conditions as reports based on biassays with pathogens usually show. Although essential oils play a part in the microbial colonization of the phyllosphere, it is not likely that inhibition of phyllosphere bacteria is essential oils’ primary role, at least in the Mediterranean environment.     

Disturbance cues in freshwater prey fishes: do juvenile convict cichlids and rainbow trout respond to ammonium as an ‘early warning’ signal?

Summary.  Several freshwater invertebrate and vertebrate prey species rely on chemosensory cues, including non-injury released disturbance cues, to assess and avoid local predation threats. The prevailing hypothesis is that a pulse of ammonia released by disturbed or stressed prey functions as the disturbance cue. Here, we test this hypothesis in two phylogenetically distant freshwater prey fishes, convict cichlids and rainbow trout. In our first experiment, we measured NH₄ ⁺ concentration in tanks containing shoals of cichlid or trout before and after exposure to a realistic model predator (or left undisturbed). We failed to find an increase in ambient NH₄ ⁺ concentration for either cichlids or trout. In our second experiment, we exposed cichlids or trout to NH₄ ⁺ at 0.1 or 0.5 mg L⁻¹ (or a distilled water control) and measured the change in antipredator behaviour (time moving, foraging rate and area use). We found no consistent increase in antipredator behaviour in response to NH₄ ⁺. In our third study, we exposed cichlids and trout to the disturbance cues of cichlids or trout (versus the odour of undisturbed donors). We found significant increases in antipredator behaviour, regardless of donor species, for both cichlids and trout. Thus, the results of our first two experiments do not support the hypothesis that ammonium functions as a disturbance cue in prey fishes. However, the results of our final experiment do confirm the use of disturbance cues in convict cichlids and rainbow trout and support that hypothesis that the disturbance cue is indeed some generalized metabolic byproduct.     

Maternal provisioning of sequestered defensive steroids by the Asian snake <Emphasis Type="Italic">Rhabdophis tigrinus</Emphasis>

Summary.   Rhabdophis tigrinus obtains defensive steroids (bufadienolides) from its diet and sequesters those compounds in specialized structures on its neck known as nuchal glands. Hatchling snakes lacking these steroids must acquire them from toads consumed as prey. Here we show that females provision bufadienolides to their offspring in amounts correlated to the quantity in their own nuchal glands; thus, chemically protected mothers produce defended offspring. Bufadienolides can be provisioned to embryos via deposition in yolk and by transfer across the egg membranes within the oviducts. Maternally provisioned bufadienolides persist in the nuchal glands of juvenile snakes from the time of hatching in late summer until the following spring, when toads of ingestible size become abundant. Therefore, maternal provisioning may provide chemical protection from predators for young R. tigrinus in the absence of dietary sources of bufadienolides.     

Uptake and toxicity of some pesticides on three freshwater fishes (Oreochromis niloticus,Clarias gariepinus and Chrysicthys nigrodigitatus)

Studies were carried out to determine the toxicity of some selected pesticides on fresh water fish in a tropical environment. The uptake of the pesticides lindane, pentachlorophenol (PCP), and propoxur, which are frequently used by farmers and industrialists were studied in concrete ponds at the University of Cape Coast, in Ghana. The fishes used for the study were Oreochromis niloticus, Clarias gariepinus, and Chrysicthys nigrodigitatus, and were obtained from cultured ponds in the Cape Coast District and Mankessim in the Central Region and Weija Dam, in the Greater Accra region of Ghana. Single high lethal concentration (SD) or acute treatment and cumulative/chronic (or multiple minor) lethal concentration (CD) treatment were employed in administering the pesticides to the fishes via water. Gas chromatograph (GC) Electron Capture Detector (ECD) analysis was done on the dead fishes to see the extent of ingestion. The LC₅₀ values obtained for lindane on the three fish samples were as follows: Chrysicthys?–?0.38?mg?L^?1; Oreochromis?–?0.42?mg?L^?1, and Clarias?–?1.2?mg?L^?1. Mortalities occurred in fish within 3–5 days of application. For the PCP on Chrysicthys, Oreochromis, and Clarias species the LC₅₀ values were 0.42, 0.32 and 0.64?mg?L^?1, respectively for over a 2–3 day period. For a three-time influx period of propoxur the LC₅₀ for Chrysicthys, Oreochromis, and Clarias, were 22.0, 30.40, and 45.04 (all in mg?L^?1), respectively. The results obtained indicated that the pesticides had adverse effects on the general growth and reproduction of fishes. Gonadosomatic indices also showed that the pesticides affected the development of the body, the gonads, and their reproduction.     

Glucosinolates and other metabolites in the leaves of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> from natural populations and their effects on a generalist and a specialist herbivore

Because many secondary metabolites in plants act as defense against herbivores it has been postulated that these compounds have evolved under selective pressure by insect herbivores. One explanation for the within-species variation in metabolite patterns in a particular species is that different populations are under selection by different herbivores. We tested this hypothesis, using Arabidopsis thaliana plants that originated from dune and inland areas. We analyzed Arabidopsis thaliana leaves using NMR spectroscopy and multivariate data analysis. Major differences in chemical composition were found in water-methanol fractions and were due to higher concentrations of sinigrin and fumaric acid in dune plants. Inland plants showed lower levels of glucose. Quantitative analysis of glucosinolates was performed with HPLC. Individual plants and populations demonstrated differences in glucosinolate composition and concentration. In growth chamber experiments, the generalist herbivore, Spodoptera exigua grew significantly better on the inland plants, while the specialist herbivore Plutella xylostella performed equally well on plants of both origins. Aliphatic glucosinolate as well as total glucosinolate concentrations negatively correlated with larval mass of Spodoptera exigua. No significant correlations, however, were found between larval mass of Plutella xylostella and glucosinolates in the leaves. A specialist and a generalist herbivore were responding differently to plant secondary chemistry, as was also found in several other studies. This is an important indication that differences in glucosinolate concentrations among populations may result from differential selection by different guilds of herbivores.     

First steps in strategic environmental assessment (SEA) adoption for coastal planning in Sicily (South Italy)

The Industrial District (I.D.) of Messina was the first public authority to consider useful the application of the Strategic Environmental Assessment (SEA) procedure for the development of an Industrial Plan regarding a coastal area. At the moment the I.D. includes six areas, but the application of the SEA concerns only the area of Pace del Mela (along the north-eastern coast of Sicily), where a “District of the two Seas” will be created, based on the synergetic development between an industrialized niche (shipbuilding) and touristic development (port and hotels). The area in which this type of intervention will occur is not in use at the moment and it was not possible to collect environmental data. For this reason, in order to describe the environmental features of this territory, we took into consideration a few indicators which could be easily identified, necessary for a post evaluation which considers the Environmental Impact Assessment (EIA) of the industrial and suburban structures to be built.     

Use of landscape and land use parameters for classification and characterization of watersheds in the mid-Atlantic across five physiographic provinces

The Atlantic Slope Consortium (ASC) is a project designed to develop and test a set of indicators in coastal systems that are ecologically appropriate, economically reasonable, and relevant to society. The suite of indicators will produce integrated assessments of the condition, health and sustainability of aquatic ecosystems based on ecological and socioeconomic information compiled at the scale of estuarine segments and small watersheds. The research mandate of the ASC project is the following:

Using a universe of watersheds, covering a range of social choices, we ask two questions:

• ? How “good” can the environment be, given those social choices?
• ? What is the intellectual model of condition within those choices, i.e., what are the causes of condition and what are the steps for improvement?

As a basis for compiling ecological indicators, a watershed classification system was required for the experimental design. The goal was to develop approximately five categories of watersheds for each physiographic province, utilizing landscape and land use parameters that would be predictive of aquatic resource condition. All 14-digit Hydrologic Unit Code (HUC) watersheds in the Mid-Atlantic region would then be classified according to the regime. Five parameters were utilized for the classification: three land cover categories, consisting of forested, agricultural, and urban, median slope or median elevation, and total variance of land covers in 1-km-radius circles positioned on all stream convergence points in a specified 14-digit?HUC watershed. Cluster analysis utilizing these five parameters resulted in approximately five well-defined watershed classes per physiographic province. The distribution of all watersheds in the Mid-Atlantic region across these categories provides a unique report on the probable condition of watersheds in the region.     

Ecologically neglected tannins and their biologically relevant activity: chemical structures of plant ellagitannins reveal their in vitro oxidative activity at high pH

Summary.  Chemical structures of 27 ellagitannins were systemically compared in respect of their in vitro oxidative activity at high pH found e.g. in lepidopteran insects. The analysis revealed over six-fold differences in the oxidative activities of individual ellagitannins which could be explained by the chemical divergences of the ellagitannins. These findings allowed the formulation of a simple equation that can be used to estimate the oxidative activities of other ellagitannins with known structures. The results suggest that, in future studies of plant-herbivore interactions, ellagitannins should be (1) taken into account as possible oxidative stress -based defences of plants against herbivores, (2) chemically characterized from the study plants, and (3) quantified individually, not as chemically ill-defined group. These actions together with the utilization of the created equation would allow the clarification of the role of ellagitannins in plant-herbivore interactions as natural pro-oxidants.     

Sex pheromone of <Emphasis Type="Italic">Ectinus aterrimus</Emphasis> (L<Emphasis Type="SmallCaps">inné</Emphasis>, 1761) (Coleoptera: Elateridae)

Summary.   Ectinus aterrimus (L.) is a fairly common European click beetle species which develops mainly in forests. In pheromone gland extracts of female E. aterrimus, examined using gas chromatography – mass spectrometry (GC-MS), one single compound was present. This was identified as 7-methyloctyl 9-methyldecanoate by comparison with a synthetic sample. Field trapping trials revealed a highly significant attraction of male E. aterrimus towards this ester. The structure of the compound differs remarkably from the pheromones of the closely related Agriotes spp., which exclusively use terpene esters.     

Sesquiterpene Hydrocarbons: Kairomones with a releaser effect in the sexual communication of the white-spotted longicorn beetle, <Emphasis Type="Italic">Anoplophora malasiaca</Emphasis> (Thomson) (Coleoptera: Cerambycidae)

Summary.  We conducted a series of experiments with the white-spotted longicorn beetle Anoplophora malasiaca (Thomson), and its host plant, Citrus unshiu, to examine the origin of the sesquiterpene hydrocarbons (SHCs; including β-elemene, β-caryophyllene, α-humulene, α-farnesene, and several unidentified compounds) that are contained in the elytra of the beetles and act as an attractant. In the laboratory, mechanically wounded citrus branches, as well as those fed upon by A. malasiaca, attracted males more frequently than intact branches. Solid phase microextraction (SPME) and subsequent analyses by gas chromatography (GC) analyses detected measurable SHCs from the air around both mechanically wounded and beetle-infested branches, as well as trace amounts from intact branches. The SHCs were also detected for a certain time from beetles that had fed on the citrus branches, but the amounts decreased rapidly after they were removed from the host. This decrease generally corresponded with a reduction of attractiveness of the beetles in a behavioural assay. Isolated females acquired the SHCs after exposure to, but not upon contact with, other females that had fed on C. unshiu branches. We hypothesize that the citrus SHCs are adsorbed in, retained on, and released from the wax layer of the beetle elytra. Since these compounds are released from branches when beetles feed, they may indirectly signal the presence of beetles to others in the field. The high response rate to SHCs by males is likely representing mate searching behavior. The SHCs act as kairomones with a releaser effect in the communication system of A. malasiaca. Hiroe Yasui, Toshiharu Akino, Midori Fukaya: These authors contributed equally to this article.     

Intraguild predation by <Emphasis Type="Italic">Harmonia axyridis</Emphasis> on coccinellids revealed by exogenous alkaloid sequestration

Summary.  Under laboratory conditions, the multicolored Asian lady beetle, Harmonia axyridis is well known as an intraguild predator of other ladybirds. However the real impact of this exotic species on native species was poorly investigated in the field. Because many ladybird species produce alkaloids as defensive compounds, we propose here a new method of intraguild predation monitoring in coccinellids based on alkaloid quantification by GC-MS. In laboratory experiments, adaline was unambiguously detected in fourth instar larvae of H. axyridis having ingested one egg or one first instar larva of Adalia bipunctata. Although prey alkaloids in the predator decreased with time, traces were still detected in pupae, exuviae and imagines of H. axyridis having ingested one prey when they were fourth instar larvae. Analysis of H. axyridis larvae collected in two potato fields shows for the first time in Europe the presence of exogenous alkaloids in 9 out of 28 individuals tested. This new method of intraguild predation detection could be used more widely to follow the interactions between predators and potential chemically defended insect preys.     

Effects of a phycotoxin,okadaic acid,on oyster heart cell survival

Okadaic acid (OA) is a dinoflagellate toxin which accumulates in shellfish producing diarrhetic shellfish poisoning (DSP) in humans. It was found that OA is a highly selective inhibitor of protein phosphatase types 1 (PP1) and 2A (PP2A) which produces a marked increase in phosphorylation of several proteins, including p38 mitogen-activated protein (MAP) kinase. The cytotoxicity attributed to OA and the effects on p38 MAP kinase and calcium current were examined in the oyster Crassostrea gigas in this study. Data showed that p38 MAP kinase is strongly expressed in oyster heart and that OA bioaccumulated in cultured heart cells. Hence the effects of OA was tested in vitro and in vivo on oysters. OA was found to (i) exert a positive chronotropic effect on cultured atrial cardiomyocytes which is related to an increase in calcium current via PKC as shown by patch clamp measurements, (ii) produce an activation/phosphorylation of MAP kinase as shown by Westernblot while the non-phosphorylated p38 remained constant during treatment, (iii) did not induce a pro-apoptotic effect. Data suggest that OA may also stimulate the anti-apoptotic pathway by phosphatase inhibition.     

“Anting” in Blue Jays: evidence in support of a food-preparatory function

Summary.  Anting, the plumage-dipping behavior to which ants (mostly formicines) are commonly subjected by birds (mostly passerines), is shown in tests with hand-raised Blue Jays (Cyanocitta cristata) and the ant Formica exsectoides to be instinctive: the birds displayed typical renditions of the behavior on the first occasion that they encountered ants. Evidence is presented supportive of the view that anting is a strategy by which birds render ants fit for ingestion. Formicine ants are ordinarily protected by their formic acid-containing spray. Being wiped into the bird’s plumage causes them to discharge that spray, without harm to the bird, to the point of almost total emptying of the glandular sac in which the secretion is stored. The ants are therefore essentially secretion-free by the time they are swallowed. Further evidence indicates that it is the ant’s possession of the acid sac that triggers the anting behavior in the bird. If F. exsectoides are surgically deprived of their acid sac, they are eaten by the birds without first being subjected to anting. Data are also presented indicating that the ant’s crop, which is especially capacious in formicines (its contents may amount to over 30% of the formicine’s mass), and which appears to survive the anting procedure intact, constitutes, at least when laden, a valuable component of the trophic package that the bird accesses by anting.     

Effects of UV-B radiation on the gills of Catla catla during early development

Scanning electron microscopic (SEM) study of gills of Catla catla catla (17-day-old) exposed to UV-B radiation (145?µW?cm^?2 at the water surface) for three different exposure times: 5, 10 and 15?min was conducted. Fish without UV-B exposure served as control. UV-B radiation damaged both gill filaments and lamellae. The intensity of damage was minimal in 5?min exposed fish, followed by 10?min exposed fish and maximal in 15?min. The gill epithelium was severely damaged in 15?min irradiated fish compared to control. Pavement cells (PVCs) were damaged and the numbers of microridges within PVCs decreased. The deep boundary of PVC was not clear. In some area of gill epithelium, PVCs were destroyed and mitochondrion-rich cells (MRCs) were exposed. The 5?min exposure reduced the number of microridges in the PVCs, but the boundary of PVCs was still visible. MRCs in the gill epithelium were not exposed in 5?min exposed fish. The damage to PVCs and subsequent exposure of MRCs in UV-B irradiated fish may hamper respiratory functions and disturb osmoregulation in catla.     

Differential activity of non-fluorinated and fluorinated analogues of the European corn borer pheromone

Summary.  The differing antagonist activity of (Z)-13-hexadecen-2-one (Z11 – 14 :MK, 1) and its 1,1,1-trifluoro derivative (Z11 –14 :TFMK, 2), two closely related analogues of the European corn borer pheromone Ostrinia nubilalis (Z strain), and their rationale is reported. Both chemicals exhibited some electrophysiological activity, and topical application of 10 pg of pheromone analogue on male antennae was sufficient to induce significantly lower depolarization responses to the pheromone versus untreated insects. In a wind tunnel, the number of European corn borer males attracted to sources containing mixtures of 1 + pheromone in ratios ≥ 1 :1 was significantly lower than the number attracted to a source containing pheromone alone. Source contact behaviour was dramatically impaired when the 1 + pheromone blend reached a ratio of 10 :1, in which only 2% of males displayed source contact in the presence of antagonist. When compound 1 was present at the source, males usually flew upwind with occasional downwind reversals; when compound 2 was present at the lure, males performed wider crosswind reversals, with little progress toward the source. In the field, traps baited with mixtures of both compounds with the pheromone in ratios of 5 :1 and 10 :1 elicited a significantly decreased number of male catches. In esterase inhibition assays, compound 2 was a potent inhibitor (IC₅₀ = 70 nM), whereas the non-fluorinated compound 1 was not. The different activity of both compounds is presumed to be due to different mechanisms of action; considerations for using methyl ketone analogues as new behavioural antagonists of the pheromone are outlined.     

Geochemical and mineralogical characteristics of elephant geophagic soils in Udawalawe National Park,Sri Lanka

Geophagy or deliberate ingestion of soils was observed among Asian elephants (Elephas maximus) in the Udawalwe National Park, Sri Lanka, for several years. The geochemical and mineralogical composition of the clayey soil layers which are purposefully selected and eaten by elephants in the park were studied, in order to identify the possible reasons for elephant geophagy. The concentrations of major and trace elements were determined by means of X-ray fluorescence spectrometry in 21 soil samples from eight geophagic sites and six soil samples collected from four non-geophagic sites. The mineralogical composition of selected soil samples was investigated using X-ray diffractometry (XRD). These geochemical analyses revealed that geophagic soils in the study areas are deeply weathered and that most of the elements are leached from the soil layers under extreme weathering conditions. The XRD data showed that the soils of the area consisted mainly quartz, feldspar, and the clay minerals kaolinite, Fe-rich illite, and smectite. Although no significant geochemical differences were identified between geophagic and non-geophagic soils, a clear difference was observed in their clay mineralogical content. Soils eaten by elephants are richer in kaolinite and illite than non-geophagic soils, which contain a higher amount of smectite. It is suggested that elephants in Udawalawe National Park ingest soils mainly not to supplement the mineral contents of their forage but to detoxify unpalatable compounds in their diet.     

Does defoliation induce chemical and morphological defenses in the leaves of silver birch seedlings under changing climate?

Summary.  We examined the effects of defoliation con-currently with elevated temperature and CO₂ on some chemical and morphological characteristics in the leaves of silver birch seedlings (Betula pendula). We also analyzed the consequent changes in the palatability of leaves for adult blue alder leaf beetles (Agelastica alni). Under the different climatic treatments, the seedlings were subjected to three fertilizer treatments (0 kg, 130 kg and 270 kg N ha⁻¹) and defoliation treatments (0%, 25% and 50% of the total leaf area). In each climatic treatment, fertilization increased the nitrogen content in the leaves, but decreased total concentrations of soluble phenolics, detected by high-performance liquid chromatography (HPLC), and insoluble condensed tannins. Defoliation, both independently and in combination with elevated temperature and CO₂, decreased the concentrations of the phenolics. Compared to the intact controls, the leaves of the defoliated seedlings were smaller and tougher. Under elevated temperature, the beetles consumed a smaller amount of the leaves of plants subjected to the high fertilization, while under ambient climatic conditions, fertilization increased the feeding. The total leaf consumption was higher under the ambient climatic conditions than under elevated temperature, elevated CO₂ or the combination of elevated temperature and CO₂.