Learned food aversion with and without an odour cue for protecting untreated baits from wild mammal foraging

Lethal methods of controlling wildlife foraging damage may present difficulties regarding conservation, ethics, welfare, and efficacy, providing an imperative to develop non-lethal alternatives. Learned food aversions offer a benign, targeted alternative to culling. However, food aversions are more readily associated with taste than with non-taste cues, and successful management applications have been limited primarily to reducing (bird) damage to treated items. To determine whether untreated foods might be protected without becoming damaged through sampling (tasting), we tested the role of an odour cue in learned food aversions with wild mammals using three conditioning treatments: (a) a combination of ziram (aversive agent) and clove oil (novel odour); (b) ziram; (c) clove oil. We tested two predictions: (1) ziram will produce learned aversion towards untreated foods; (2) training with the ziram–clove combination will produce enhanced aversion towards untreated foods in the presence of a clove odour cue. We collected detailed behavioural data from free-ranging animals using video surveillance. With European badgers Meles meles, both ziram treatments invoked innate aversion to baits. Ziram did not produce learned aversion to untreated foods, but conditioning badgers with the ziram–clove combination produced learned aversion towards untreated foods in the presence of the odour cue. The clove control did not affect badger behaviour. Our results support the second, but not the first, prediction with badgers. Red foxes Vulpes vulpes attended the ziram and the clove treatments only. Foxes displayed innate aversion to ziram, and subsequently developed learned aversions to untreated baits. Our results with foxes therefore support the first prediction, but we were unable to test the second. Learned food aversions using odour cues could potentially provide non-lethal protection for untreated foodstuffs, such as eggs, crops, and fruit, from mammalian pest species.     

Predator pheromone elicits a temporally dependent non-consumptive effect in prey

1. The influence of predator cues on the behaviour of prey is well supported in the literature; however, a clear understanding of how predator cues affect prey in variable environmental conditions and over longer time scales is needed to better understand the underlying mechanisms. Here, we measure how predator odors affect herbivore colonization, abundance, oviposition, and plant damage across two growing seasons.
2. The study system consisted of Leptinotarsa decemlineata (Colorado potato beetle) as prey, and the aggregation pheromone of live Podisus maculiventris (spined soldier bug) as the predator cue in a potato field.
3. In 2016, the amount of feeding damage by early beetle colonists was lower in predator odor-treated plots, reducing plant damage by 22%. Larval abundance was also reduced in treated plots in 2016. Beetle abundance and damage in 2017 was similar in the treatment and control plots. Two mechanisms were investigated to better understand why prey response to the predator odor treatment weakened over the first season, including changes in predator odor cue strength and prey habituation. Predator odor cue strength emerged as a likely explanation, as dispensers, which released a synthetic predator pheromone over the entire season, reduced the probability of finding damage more consistently than the live predator treatment.
4. These results suggest that temporal patterns of predator cue release and strength may drive prey response across the season, underscoring the importance of cue release-rate and consistency in both species interactions and for the future application of modifying insect behaviour using non-consumptive effects in agricultural systems.

     

Clove Oil Efficacy on the Red Spider Mite, <Emphasis Type="Italic">Oligonychus coffeae</Emphasis> Nietner (Acari: Tetranychidae) Infesting Tea Plants

The tea red spider mite, Oligonychus coffeae Nietner (Tetranychidae), is an economically important pest of agricultural and ornamental crops and considered one of the major pests of tea plants in North-east India. In view of increasing resistance recorded in insect and mite pests against pesticides, a study was conducted to determine the acaricidal, antiovipositional, repellent and ovicidal activities of clove oil (an essential oil from the clove plant, Syzygium aromaticum L. Merr. & Perry: Myrtaceae) against tea-red-spider-mite. Mortality of O. coffeae varied with the concentrations and the duration of exposure time of the mites after application of oil. Rate of deposition of eggs by mites on treated leaf surfaces as well as the viability rate of eggs decreased significantly. In addition, certain concentrations of clove oil have been proved effective against adult mites.     

Predator Recognition and Learned Irrelevance in the Crayfish Orconectes virilis

When individuals of the crayfish Orconectes virilis detect an unlearned danger cue (alarm odor) and a novel cue (goldfish odor) at the same time, they form a learned association and behave as if the novel cue is associated with increased predation risk ( Hazlett et al. 2002 ). This study examined the potential for learned irrelevance in O. virilis and the circumstances under which blockage of the formation of a learned association could occur. If individuals experience a random pattern of alarm odor and goldfish odor over the days prior to the simultaneous detection of those two cues, no learned association is formed (= learned irrelevance). That is, there is no inhibition of responses to a food cue when goldfish odor is added if the crayfish has experienced a random pattern of the two cues. Learning was eliminated if the random pattern of cues was experienced before or after the simultaneous detection. To present the two cues (alarm and goldfish odors) to crayfish independently on separate days, the water containing goldfish odor had to be removed from the aquaria as the odor persisted at least 24 h. The importance of the learned irrelevance phenomenon on predator–prey interactions is discussed.     

Conspecific olfactory preferences and interspecific divergence in odor cues in a chickadee hybrid zone

Understanding how mating cues promote reproductive isolation upon secondary contact is important in describing the speciation process in animals. Divergent chemical cues have been shown to act in reproductive isolation across many animal taxa. However, such cues have been overlooked in avian speciation, particularly in passerines, in favor of more traditional signals such as song and plumage. Here, we aim to test the potential for odor to act as a mate choice cue, and therefore contribute to premating reproductive isolation between the black‐capped (Poecile atricapillus) and Carolina chickadee (P. carolinensis) in eastern Pennsylvania hybrid zone populations. Using gas chromatography–mass spectrometry, we document significant species differences in uropygial gland oil chemistry, especially in the ratio of ester to nonester compounds. We also show significant preferences for conspecific over heterospecific odor cues in wild chickadees using a Y‐maze design. Our results suggest that odor may be an overlooked but important mating cue in these chickadees, potentially promoting premating reproductive isolation. We further discuss several promising avenues for future research in songbird olfactory communication and speciation.     

The effects of clove oil on coral: An experimental evaluation using Pocillopora damicornis (Linnaeus)

Clove oil solution (10% clove oil, 90% ethanol) is an anaesthetic that is widely used to catch demersal fish on coral reefs. This study assessed the effects of clove oil solution on colonies of Pocillopora damicornis, a cosmopolitan reef coral. In the laboratory, low concentrations (0.5 ppt) of clove oil solution had no effect on coral colour or photosynthetic efficiency, irrespective of exposure time (1-60 min). Corals treated with high concentrations (50 ppt) of clove oil solution died immediately, including those that were exposed briefly (1 min). Intermediate concentrations (5 ppt) of clove oil solution produced variable results: a 1 min exposure had no effect, a 10 min exposure caused bleaching and reduced photosynthetic efficiency, and a 60 min exposure caused total mortality. To validate these observations, clove oil solution was applied to corals in situ. Sixty-three days after application, corals treated with 10 ml of clove oil solution appeared to be unaffected. It was concluded that (1) limited amounts of clove oil solution are unlikely to harm this coral, and (2) clove oil solution may represent an ‘eco-friendly’ alternative to cyanide for use in the live reef-fish trade.     

Reports on badgers Meles meles in Dutch newspapers 1900–2013: same animals,different framings?

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Alliin lyase localization in bundle sheaths of the garlic clove (Allium sativum)

The enzyme alliin lyase (E.C. 4.4.1.4) catalyzes formation of allicin, the parent of several sulfur-containing compounds responsible for flavor, odor, and pharmacological properties of garlic (Allium sativum). Alliin lyase is a major product of the storage bud (clove), accounting for 10% of its total protein. Accumulation of this protein was characterized by locating alliin lyase deposits within the clove. Paraffin sections stained for general protein using aniline blue-black reveal dense deposits within parenchymatous bundle sheaths. Deposits are most pronounced around phloem. Remaining storage parenchyma, not in contact with bundles, appears structurally uniform, with some protein accumulating in cells near the outer surface of the clove. In freehand sections of unfixed cloves, bundle sheath cells are the only ones to show green autofluorescence when excited by blue light. Such fluorescence is consistent with the presence of pyridoxal phosphate cofactor of alliin lyase. An alliin lyase activity stain, based on detecting aminocrotonate-generating enzymes, shows activity to be restricted to bundle sheath cells in fresh material. Finally, enrichment of alliin lyase in bundle sheaths is shown by immunocytochemical staining of these areas using a polyclonal antibody generated against purified enzyme. Aliin lyase concentrates in bundle sheaths, while little if any occurs in storage mesophyll not in contact with vascular bundles. Deposits in the cloves may reflect the enzyme's role in protecting underground storage buds from decay and predation. Positioning near the phloem suggests that alliin lyase, or compounds related to its metabolism, may be translocated to and from the clove during development.     

The effect of background plant odours on the behavioural responses of Frankliniella occidentalis to attractive or repellent compounds in a Y‐tube olfactometer

The western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), causes major losses in agricultural and horticultural crops worldwide. The volatile compounds methyl isonicotinate, p‐anisaldehyde, eugenol, and linalool are known as olfactory attractants, and salicylaldehyde is known as a repellent for F. occidentalis under clean‐air conditions in laboratory experiments. In the present study we assessed the responses of F. occidentalis to these compounds when presented alone, in combination, and in the presence of background odours emanating from cucumber (Cucumis sativus L., Cucurbitaceae), capsicum (Capiscum anuum L., Solanaceae), chrysanthemum (Chrysanthemum morifolium Ramat., Asteraceae), clove basil (Ocimum gratissimum L.), and lavender (Lavandula angustifolia L., both Lamiaceae) plants in a Y‐tube olfactometer. In the presence of any background plant odour, the attractiveness of pure methyl isonicotinate to F. occidentalis proved to be consistently significant. Compared to clean‐air conditions, a slightly lower percentage of thrips chose the Y‐tube arm loaded with 10% p‐anisaldehyde in the presence of cucumber leaf odour. With non‐flowering clove basil plants in the background, F. occidentalis responses to 1% eugenol, a constituent of clove basil essential oil, were neutral, and the same applied to responses to pure linalool, a constituent of lavender essential oil, in the presence of flowering lavender plants. Also, thrips responses to pure or diluted salicylaldehyde were clearly influenced by plant background odours. We simulated a push‐pull situation and found a trend indicating that the percentage of F. occidentalis choosing the airflow loaded with the attractant methyl isonicotinate was higher when the airflow in the other arm of the Y‐tube was loaded with the repellent salicylaldehyde compared to clean air, and vice versa. We showed interactions between attractive or repellent volatile compounds and the environmental odours in the chemical ecology of F. occidentalis and the potential of a combined use of these compounds in thrips pest management.     

Effects of the fish anesthetic, clove oil (eugenol), on coral health and growth

Ecological research within coral reefs often requires the use of anesthetics to immobilize organisms. It is therefore important to consider the effect of these chemicals on the surrounding flora and fauna, particularly to the corals themselves. We quantified the effects of clove oil, a commonly used fish anesthetic, on the growth and occurrence of bleaching in three species of corals: Acropora striata, Pocillopora verrucosa, and Porites australiensis. We compared coral responses to five treatments: a gradient of four clove oil concentrations (0-28%) in seawater, and one concentration of clove oil (14%) in ethanol. Each week, we assessed the presence of bleaching, and then applied the treatment. We measured growth over the duration of the 6-week experiment using the buoyant weight technique. Growth and bleaching showed a dose response to clove oil exposure, and the use of ethanol as a solvent had an additional deleterious effect, as also suggested by observed changes in concentrations of eugenol following field application. Overall, growth was reduced by 37.6% at the highest concentration (28% clove oil in seawater) relative to the control (0% clove oil). The reduction in growth was nearly as great (35.3% of the control) at half the concentration of clove oil (14%) when dissolved in ethanol. These results suggest the repeated use of clove oil (even without a solvent) can deleteriously affect corals.     

Repellent activity of essential oils on adults of Halyomorpha halys (Heteroptera: Pentatomidae) in different physiological-behavioural phases

The brown marmorated stink bug Halyomorpha halys is an invasive agricultural pest in North America and Europe, and also a dwelling nuisance in autumn, due to the overwintering adults aggregating inside buildings. The repellent potential of ginger, clove, vetiver and turmeric essential oils (EOs) was tested on H. halys adults of three different physiological-behavioural phases: exiting overwintering (EXOV), active during summer (SUMM) and entering overwintering (ENOV). In a two-choice apparatus, fresh food was used as an attractant in both sides, together with three males when testing ENOV individuals. A filter paper with different concentrations of the EOs was the treatment side, and a filter paper with ethanol acted as control. The position of the individually tested bugs was recorded after 1, 6 and 24 hr. All tested EOs were repellent at concentrations higher than 3%, independent of sex or length of exposure. Turmeric and clove were the most repellent EOs, whereas ginger and vetiver showed on average a medium-to-low repellency. Significant differences emerged among the physiological-behavioural phases, with SUMM individuals showing a greater repellency to many of the tested concentrations, and EXOV individuals being overall the least susceptible to these substances. The response to vetiver oil was ambiguous, as at 25%, it elicited both a strong repellency in SUMM and a strong attraction in EXOV. Turmeric and clove EOs are promising candidates in integrated pest management strategies to reduce attacks by H. halys to susceptible crops especially during summer, as well as to prevent the entrance of overwintering bugs in buildings in autumn.     

四种植物精油对黑翅土白蚁触杀和驱避作用

为筛选出高效防治黑翅土白蚁的天然植物精油,减少有机合成农药的使用,该文研究了大蒜精油、肉桂油、丁香油和印楝素油四种植物精油对黑翅土白蚁的触杀效果和驱避作用。结果表明:大蒜精油、肉桂油和丁香油的浓度为5和10 mg·m L~(-1)时,处理2 h后,黑翅土白蚁的校正死亡率达100%,而相同浓度的印楝素油和对照处理的黑翅土白蚁校正死亡率低于5%。随着处理时间延长,浓度为1.25和2.5 mg·m L~(-1)的大蒜精油、肉桂油和丁香油处理6 h时,黑翅土白蚁的校正死亡率仍达100%,而此时对应的印楝素油和对照处理的黑翅土白蚁校正死亡率仅为10%,说明大蒜精油、肉桂油和丁香油对黑翅土白蚁具有较强的触杀效果。大蒜精油、丁香油和肉桂油在处理黑翅土白蚁2 h后LC_(50)值(半致死量)分别为1.572、1.05和1.03mg·m L~(-1),说明肉桂油对黑翅土白蚁的毒性相对最大,触杀效果最好。此外,10 mg·m L~(-1)的大蒜精油、肉桂油、丁香油和印楝素油的驱避试验表明,处理4、6、8和12 h后,大蒜精油、肉桂油和丁香油三精油处理区的黑翅土白蚁数均显著低于对照区的,驱避率总体93%,而对应的印楝素油的驱避率总体28.5%,表明大蒜精油、丁香油和肉桂油三种植物精油对黑翅土白蚁均有显著的驱避活性。综上可知,四种植物精油中大蒜精油、肉桂油和丁香油在防治黑翅土白蚁方面应用潜力很好,是开发绿色环保白蚁防治药剂的可选材料。     

Antibacterial Potent of Acetylated and Non-Acetylated Clove Bud Essential Oils and Their Main Compounds

Clove bud is a medicinal plant used traditionally in Asia for the treatment of various disease. Previously, Clove oil is a potential source of an antimicrobial compounds especially vis-a-vis bacterial pathogens. However, the compound responsible for this activity remains to be investigated. Essential oil (EO) clove, acetylated essential oil clove, eugenol, and acetyleugenol were evaluate as an antibacterial potential agent against Staphyloccocus aureus (SE), Escherichia coli (EC) and Pseudomonas aeruginosa (PA). Essential oil containing eugenol was extracted from buds of Eugenia caryophyllata commonly named clove (Syzygium aromaticum (L.) (Family Myrtaceae) by a simple hydrodistillation. The analysis of the essential oils (EOs) using gas chromatography-mass spectrometry (GC-MS) shows eugenol as the major constituent with 70.14 % of the total. The Eugenol was isolated from the EO using chemical treatment. Afterwards, the EO and eugenol were converted to acetylated EO and acetyleugenol, respectively using acetic anhydride. The antibacterial result revealed that all compounds showed a strong activity against the three strains. The Staphyloccocus aureus and Pseudomonas aeruginosa were extremely sensitive against eugenol with an inhibition diameters of 25 mm. The MIC values of eugenol versus S. aureus and P. aeruginosa were 0.58 and 2.32 mg/mL, respectively, while the MIB values were 2.32 mg/mL and 9.28 mg/mL.     

In vitro fungitoxicity of the essential oil of Syzygium aromaticum

The in vitro antifungal activity of clove oil was studied against four test fungi namely Alternaria alternata, Fusarium chlamydosporum, Helminthosporum oryzae and Rhizoctonia bataticola by the agar well diffusion method. These test fungi were found to be highly sensitive to clove oil at a concentration of 100 μl/well. The inhibition zone diameter was found to be in the range of 55–65 mm. The toxicity of clove oil on the germination and growth of A. alternata was further examined in liquid medium. Concentration- and time-dependent toxicity was recorded from 0.05 to 20% (v/v) concentration. The minimum fungistatic concentration was found to be 0.05%. Above this concentration, lysis of conidia and inhibition of mycelial growth were detected. Microscopic analysis showed 20–40% lysis of conidia after 72 h of incubation at 5% concentration. However at higher clove oil concentration (10%), up to 20% of conidia were lysed within 24 h of incubation. Similar concentration- and time-dependent toxicity was observed at different concentrations and time intervals. The findings indicated that clove oil possesses fungicidal activity against phytopathogenic fungi. Further study is required to determine whether it could have value in the management of plant infectious diseases. This revised version was published online in July 2006 with corrections to the Cover Date.     

Testing clove oil as an anaesthetic for long-distance transport of live fish: the case of the Lake Victoria cichlid Haplochromis obliquidens

Clove oil can be used as an anaesthetic in the handling of marine and freshwater fish. Few studies report on its use for periods up to 48 h, for example, under long‐distance transport conditions. This study tested the effect of different clove oil concentrations for 1–48 h on recovery and survival of the cichlid Haplochromis obliquidens, an ornamental fish species endemic to Lake Victoria. Haplochromis obliquidens were anaesthetized for 1 h using 5–25 μl L^?1 clove oil. There was no correlation between clove oil concentration and post‐anaesthesia recovery time (P = 0.15). On average, fish recovered within 9.5 ± 2 min, and no fish died within 24 h after recovery. Results from exposure of fish to 18–20 μl L^?1 clove oil for up to 48 h suggested a narrow margin of safety as this concentration range induced mortality. At 18 μl L^?1 recovery times ranged from 3 to 43 min between 24 and 36 h exposure, while fish exposed longer than 36 h recovered within 1–10 min, or within 1–2 min after 44–48 h. At the end of a 48‐h transport experiment total ammonia levels were higher in transport water containing anaesthetized fish than for non‐anaesthetized fish (1.65 ± 0.19 and 0.54 ± 0.08 mg L^?1 NH + NH₃, respectively). The combined use of clove oil and the selective ammonium ion exchanger zeolite was considered feasible as ammonia levels could be reduced by up to 82% compared to control bags without zeolite.     

Acaricidal activity of essential oil of Syzygium aromaticum,hydrolate and eugenol formulated or free on larvae and engorged females of Rhipicephalus microplus

The cattle tick, Rhipicephalus microplus (Canestrini, 1888) (Ixodida: Ixodidae), is the most important ectoparasite in cattle‐breeding areas and is responsible for severe economic losses. Synthetic acaricides have been used to control this parasite. However, the need for safer products has stimulated the search for new acaricides, such as those to be obtained from medicinal plants. The essential oil of Syzygium aromaticum (clove) has many biological properties and shows great potential for use in veterinary applications. In the context of the need for new agents, this study investigated the in vitro properties of the hydrolate, essential oil and the main constituent of S. aromaticum, eugenol, in formulated and free applications against larvae and females of R. microplus. Eugenol and the essential oil caused 100% mortality in larvae at starting applications of 2.5 mg/mL and 5.0 mg/mL, respectively. The hydrolate showed no activity. Both eugenol and essential oil had good efficacy in adult immersion tests at 50 mg/mL and achieved 100% efficacy at a concentration of 100 mg/mL. The results of these tests reaffirm the important potential of clove essential oil and eugenol.     

Enzymatic Modification of Palmarosa Essential Oil: Chemical Analysis and Olfactory Evaluation of Acylated Products

We have developed an enzymatic protocol to modify the composition of palmarosa essential oil by acylation of its alcohol components by three different acyl donors at various rates. The resulting modified products were characterized by qualitative and quantitative analyses by gas chromatography, and their olfactory properties were evaluated by professional perfumers. We showed that our protocol resulted in two types of modifications of the olfactory properties. The first and most obvious effect observed was the decrease of the alcohol content, with the concomitant increase of the corresponding esters, along with their fruity notes (pear, most notably). The second and less obvious effect was the expression of notes from minor components ((E)‐β‐ocimene, linalool, β‐caryophyllene, and farnesene), originally masked by the sweet‐floral‐rose odor of geraniol, present in 70% in the palmarosa essential oil used, and emergence of citrus, green, spicy and clove characters in the modified products. This methodology might be considered in the future as a sustainable route to new natural ingredients for the perfumer.     

Encoding predicted outcome and acquired value in orbitofrontal cortex during cue sampling depends upon input from basolateral amygdala

Certain goal-directed behaviors depend critically upon interactions between orbitofrontal cortex (OFC) and basolateral amygdala (ABL). Here we describe direct neurophysiological evidence of this cooperative function. We recorded from OFC in intact and ABL-lesioned rats learning odor discrimination problems. As rats learned these problems, we found that lesioned rats exhibited marked changes in the information represented in OFC during odor cue sampling. Lesioned rats had fewer cue-selective neurons in OFC after learning; the cue-selective population in lesioned rats did not include neurons that were also responsive in anticipation of the predicted outcome; and the cue-activated representations that remained in lesioned rats were less associative and more often bound to cue identity. The results provide a neural substrate for representing acquired value and features of the predicted outcome during cue sampling, disruption of which could account for deficits in goal-directed behavior after damage to this system.     

Effect of some mould inhibitors and herbal plants on mycotoxins production by Aspergillus flavus and Fusarium verticilloides in vitro and in stored corn grains

Clove oil, clove extract and butylated hydroxyanisole (BHA) completely suppressed the growth of both Fusarium verticilloides and Aspergillus flavus isolates. Black cumin and thyme extracts were more suppressive on F. verticilloides than A. flavus. Antitox-Plus (AP) had no effect on the growth of both the pathogens. The minimum inhibitory concentration (MIC) test revealed that A. flavus was more sensitive to Fix-A-Tox (FAT) and AP than F. verticilloides. In the growth media, all the tested substances, completely suppressed the production of aflatoxins by A. flavus and significantly reduced fumonisins production by F. verticilloides, particularly clove oil and extract. Treatment of immature grains with the tested mould inhibitors prior to inoculation with A. flavus and F. verticilloides significantly reduced mycotoxins production at the end of the storage period; moreover, highest reduction rates were realised by BHA and FAT. Complete or highly significant suppression of aflatoxins in mature grains were obtained by all the tested herbal and synthetic mould inhibitors. Ground clove buds contained the highest carvacrol content, whereas thymol content was higher in thyme extract. Clove oil was rich in eugenol. Alpha-tocopherol content was higher in ground black cumin (BC), followed by BC oil. Unsaturated fatty acid content was higher in thyme extract and ground BC than saturated fatty acids. Linolenic acid was the most predominant fatty acid in BC oil and extract, whereas behenic and arachidic acids were detected only in BC oil. Stearic acid was the main fatty acid in clove oil and extract, whereas oleic acid was the prevailing fatty acid in thyme extract.