Clove Buds Essential Oil: The Impact of Grinding on the Chemical Composition and Its Biological Activities Involved in Consumer's Health Security.

This study is aimed at identifying the chemical composition of the essential oil extracted from the Syzygium aromaticum seeds, as well as investigating its biological activities, insecticide effect, and allelopathic properties. The extraction yield was about 14.3 and 7.14% for grounded and ungrounded seeds, respectively. The GC-MS analysis allowed the identification of 17 heterogeneous compounds, including eugenol (68.7-87.4%), as major compound, cyperene (20.5-7.2%), phenethyl isovalerate (6.4-3.6%), and cis-thujopsene (1.9-0.8%), respectively, for grounded and ungrounded seeds. Concerning the antibacterial activity, the diameter of the inhibition zone reached 35 mm when the essential oil extracted from grounded seeds was applied against Escherichia coli. Regarding the antioxidant activity via the DPPH radical scavenging test, the IC50 varied from 1.2 ± 0.1 to 2.8 ± 0.5 µg/mL. With respect to reducing power, the efficient concentration EC50 ranged from 32 to 50 µg/mL. The essential oil exhibited also an allelopathic effect against seeds of Hyoscyamus niger, as well as an insecticide effect against Sitophilus oryzae with a DL50 value of 252.4 µL/L air. These findings enhance the use of this spice as a natural food preservative and encourage its use in several fields, including pharmaceutical, cosmetics, agriculture, and therapy, that could be a strategic way to guarantee the consumer's health.

Phytochemical profile and insecticidal activity of Agave americana leaf extract towards Sitophilus oryzae (L.) (Coleoptera: Curculionidae).

The main objective of the present study is to introduce a new and ecologically safe method for managing the rice weevil, Sitophilus oryzae. Therefore, the Agave americana leaf extract's phytochemical profile, and its insecticidal activity against the adults of S. oryzae were evaluated. The A. americana leaf extract was screened for the following phytochemicals: total phenolics (14.70 ± 0.31 mg GAE/g FW), total flavonoids (5.15 ± 0.18 mg RE/g FW) and saponins (10.32 ± 0.20 mg OAE/g FW). The HPLC-ESI/TOF-MS analysis results revealed that flavonoid glycosides (kaempferol, quercetin, and isorhamnetin derivates) were the major phenolic compounds of the A. americana leaf extract. In addition, the GC-MS analysis identified n-alkanes (77.77%) as significant compounds of the lipophilic fraction from the leaf extract. Moreover, the insecticidal potential was assessed through contact and repellent bioassays towards the rice weevil adults. The LD50, LC50, and RC50 values were 10.55 µg/insect, 8.99 µg/cm2, and 0.055 µg/cm2 for topical application method, treated filter-paper method, and repellent bioassay, respectively. Furthermore, the A. americana leaf extract inhibited digestive enzyme activities, and median inhibition concentrations IC50 were evaluated to be 146.06 ± 1.74 and 86.18 ± 1.08 µg/mL for α-amylase and protease, respectively. Overall, our results highlighted the promising potential of the leaf extract against S. oryzae adults, allowing us to recommend the extract under investigation as an ecofriendly alternative to synthetic insecticides.

Toxicity to, oviposition and population growth impairments of Callosobruchus maculatus exposed to clove and cinnamon essential oils.

The use of plant essential oils has been shown to efficiently control insect pests of stored beans, significantly reducing the threats associated with synthetic insecticides. Here, we evaluated the potential of applications of essential oils of clove, Syzygium aromaticum L., and cinnamon, Cinnamomum zeylanicum L., to control Callosobruchus maculatus, considered as one of the most cosmopolitan pests of stored beans. Using four combinations of couples (i.e., unexposed couples, exposed females, exposed males, and exposed couples), we also evaluated how sublethal exposure to these essential oils impacted C. maculatus oviposition. Bioassays results revealed that both essential oils exhibited insecticidal activities similar to the synthetic pyrethroid insecticide deltamethrin. Furthermore, oil dosage increments proportionately decreased the growth rate and reduced the losses in bean weight caused by cowpea weevils, and offspring emergence was almost abolished when parents were exposed to the LD20 of each essential oil. Finally, significant oviposition impairments were perceived only in couples where females were exposed (i.e., females exposed and exposed couples) to the LD20 of cinnamon and clove essential oils. Thus, by exhibiting similar insecticidal activities as synthetic insecticides and by significantly affecting the oviposition of sublethally exposed C. maculatus females, the cinnamon and clove essential oils represent valuable tools with potential of integration into the management of C. maculatus infestations.

Observing the devastating coffee berry borer (Hypothenemus hampei) inside the coffee berry using micro-computed tomography.

The coffee berry borer is the most devastating insect pest of coffee throughout the world. The insect spends most of its life cycle inside the coffee berry, which makes it quite difficult to observe its behaviour. Micro-computed tomography (micro-CT) was used to observe all developmental stages of the coffee berry borer inside coffee berries (Coffea canephora). An interesting oviposition pattern involving a sequential placement of eggs starting in the periphery of the seed and moving inwards was observed. Micro-CT should be useful in elucidating unknown life history aspects of other seed-feeding bark beetles as well as of bark and ambrosia beetles in general.

Role of Plant Volatiles in Host Plant Recognition by Listronotus maculicollis (Coleoptera: Curculionidae).

The annual bluegrass weevil (ABW), Listronotus maculicollis Kirby, is an economically important pest of short cut turfgrass. Annual bluegrass, Poa annua L., is the most preferred and suitable host for ABW oviposition, larval survival and development. We investigated the involvement of grass volatiles in ABW host plant preference under laboratory and field conditions. First, ovipositional and feeding preferences of ABW adults were studied in a sensory deprivation experiment. Clear evidence of involvement of olfaction in host recognition by ABW was demonstrated. Poa annua was preferred for oviposition over three bentgrasses, Agrostis spp., but weevils with blocked antennae did not exhibit significant preferences. ABW behavioral responses to volatiles emitted by Agrostis spp. and P. annua were examined in Y-tube olfactometer assays. Poa annua was attractive to ABW females and preferred to Agrostis spp. cultivars in Y-tube assays. Headspace volatiles emitted by P. annua and four cultivars of Agrostis stolonifera L. and two each of A. capillaris L. and A. canina L. were extracted, identified and compared. No P. annua specific volatiles were found, but Agrostis spp. tended to have larger quantities of terpenoids than P. annua. (Z)-3-hexenyl acetate, phenyl ethyl alcohol and their combination were the most attractive compounds to ABW females in laboratory Y-tube assays. The combination of these compounds as a trap bait in field experiments attracted adults during the spring migration, but was ineffective once the adults were on the short-mown turfgrass. Hence, their usefulness for monitoring weevil populations needs further investigation.

Susceptibility and possible resistance mechanisms in the palm species Phoenix dactylifera, Chamaerops humilis and Washingtonia filifera against Rhynchophorus ferrugineus (Olivier, 1790) (Coleoptera: Curculionidae).

Rhynchophorus ferrugineus, known as the Red Palm Weevil (RPW), is reported as a pest of up to 40 palm species. However, the susceptibility degree and the defense mechanisms of these species against this weevil are still poorly known. In Europe, the RPW is a major pest of Phoenix canariensis while other palm species, including the congeneric Phoenix dactylifera, seem to be less suitable hosts for this insect. The aim of our study was to compare the defensive response of P. dactylifera, Chamaerops humilis and Washingtonia filifera against R. ferrugineus and try to define the mechanisms of resistance that characterize these species. Bioassays were carried out to evaluate the mortality induced on RPW larvae by extracts from the leaf rachis of the studied palm species. Tests at semi-field scale were also conducted, based either on forced palm infestation, with larvae of RPW, or on natural infestation, with adult females. Rachis extracts from C. humilis and W. filifera caused 100% larval mortality after 2 days of exposure, while extracts of P. dactylifera did not impair larval survival. Independently of the effect of the leaf extracts, the weevils were unable to naturally infest the three palm species, although larval survival was high after forced infestation of the plants. We concluded that the observed lack of infestation of P. dactylifera by RPW is due to factors other than antibiosis. In W. filifera and C. humilis, although the presence of antixenosis mechanisms cannot be excluded, resistance to R. ferrugineus seems to rely on the presence of antibiosis compounds.

Overwintering, Oviposition, and Larval Survival of Hunting Billbugs (Coleoptera: Curculionidae) and Implications for Adult Damage in North Carolina Turfgrass.

The hunting billbug, Sphenophorus venatus vestitus Chittenden, is one of the most widely recognized billbug turfgrass pests. Since 2000, damage to warm-season turfgrass caused by hunting bill bugs has increased and a need for information on hunting billbug biology is necessary for the development of management plans. Field and laboratory studies were conducted to collect data on overwintering, oviposition behavior, larval survival at various levels of soil moisture, and adult damage. Turfgrass samples from 'Tifway 419' bermudagrass(Cynodon dactylon (L.) Pers x Cynodon transvaalensis Burtt Davy) on golf courses were collected to determine overwintering behavior, and 10 female adult billbugs were collected weekly to determine oviposition behavior.Survival of medium-sized larvae (head capsule width: 1.0 and 1.7 mm) was evaluated in containers with 20, 40,60, or 80% of the total pore space occupied by water. Zero, two, four, or six adult billbugs were placed in bermudagrass, zoysiagrass, or tall fescue containers and images were collected for 4 weeks to determine adult damage. We observed that hunting billbugs overwinter as adults and all larval sizes. Adults became active in March and began to oviposit, which continued through October. Larval mortality was lowest with 20% of the total pores pace occupied by water, while increases in moisture caused significant mortality. Adults caused a greater reduction in warm-season turfgrass cover than cool-season turfgrass cover. This research builds on the existing biological information for the hunting billbug biology in transition zones and will be pivotal in developing practical and sustainable management plans.

Biology and morphology of immature stages of Adosomus roridus (Coleoptera: Curculionidae: Lixinae).

Mature larva and pupa of Adosomus (s. str.) roridus (Pallas, 1781) (Curculionidae: Lixinae: Cleonini) are described and compared with ten other taxa of Cleonini with known larvae. This weevil is an oligophagous species on the Asteraceae family. From our observations in Slovakia, we found active adults from April to September in dry sunny places within extensively used or fallow vineyards and in ruderal vegetation with host plants. The weevil is a root borer--larvae, pupae and fresh adults were collected from the root necks and roots of Common Tansy (Tanacetum vulgare L.) and rarely from Mugwort (Artemisia vulgaris L.). Each plant was usually occupied by one larva, or more rarely with two or three larvae. The new generation of adult individuals appeared from early summer to autumn. Both larvae and some of the adults overwinter, which is quite unique within Cleonini.

[Influence of fruit size of Camellia meiocarpa on growth of oil tea weevil, Curculio chinensis (Coleoptera: Curculionidae)].

The relationship between mature larval mass of oil tea weevil (Curculio chinensis) and fruit volume of its host plant oil tea (Camellia meiocarpa) was fitted with Logistic equation in order to understand the restriction of host fruit size on large larval growth and development of the weevil. The results showed that the larval mass increased with the increasing host fruit volume, which was in good conformity with the Logistic model. The weevil larval growth followed the principle of diminishing marginal utility, and it could be divided into two periods, the fast-growing period (<3.216 cm3, one larva per fruit; <4.747 cm3, two larvae per fruit ) and the asymptotic growing period (>3.216 cm3, one larva per fruit; >4.747 cm3, two larvae per fruit). The minimum fruit size threshold was 1500 cm3 for one larva per fruit, and 2.539 cm3 for two larvae per fruit. The temporal pattern that the mature larvae exited from their host fruits was established, the number of larvae escaping from their host fruits decreased daily after the fruit collection, and the larval escaping peak largely appeared from 6:00 to 10:00 AM with 43.9% of total escaping number, and especially from 7:00 to 8:00 AM with 21.1% of total escaping number. The bigger the larvae, the earlier exited from their host fruits. The restriction of fruit size on growth and development of oil tea weevil was observed, and it should be a behavioral adaptation strategy to increase the offspring' s fitness for the parental weevil adults to oviposit on the bigger fruits.

Repelência e atividade inseticida de pós vegetais sobre Zabrotes subfasciatus Boheman em feijão-fava armazenado / Repellency and insecticidal activity of plant powders on Zabrotes subfasciatus (Boh. ) on stored lima beans

O objetivo desta pesquisa foi avaliar o potencial inseticida de Piper nigrum L. (pimenta do reino), Ruta graveolens L. (arruda), Laurus nobilis L. (louro), Syzygium aromaticum (L.) Merr. & L.M.Perry (cravo da índia), Chenopodium ambrosioides L. (mastruz), Piper tuberculatum Jaqc. (pimenta de macaco), Tagetes erecta L. (cravo de defunto), Cymbopogon nardus L. (citronela) e Melissa officinalis L. (erva cidreira) sobre Z. subfasciatus Boh. através dos testes de confinamento, onde os insetos foram confinados durante cinco dias em um recipiente contendo dez grãos de feijão-fava com 0,3 g do pó da planta a ser testada, e, com chance de escolha por meio de uma arena circular, os insetos tiveram a possibilidade de escolha entre grãos com os pós e a testemunha, grãos sem pó das plantas. Observou-se que houve plantas que atuaram como inseticida, outras que repeliram o inseto e não causaram a morte, e outras, que além de repelir também mataram os insetos quando em contato (cravo da índia e matruz). Concluímos que: P. nigrum, P. tuberculatum, S. aromaticum e C. ambrosioides são tóxicas à Z. subfasciatus causando-lhes a morte, L. nobilis, T. erecta, e C. nardus não apresentaram efeito tóxico sobre Z. subfasciatus, C. ambrosioides, S. aromaticum, e C. nardus são repelentes à Z. subfasciatus; P. nigrum, P. tuberculatum, e T. erecta são neutras, e que M. officinalis não apresentou nenhum efeito sobre Z. subfasciatus em relação aos parâmetros avaliados.

The purpose of this research was to evaluate the insecticidal potential of Piper nigrum L., Ruta graveolens L., Laurus nobilis L., Syzygium aromaticum Merr. & L.M.Perry., Chenopodium ambrosioides L., Piper tuberculatum Jaqc., Tagetes erecta L., Cymbopogon nardus L. and Melissa officinalis L. against Zabrotes subfasciatus by free choice, in which, by means of a circular arena, insects had a choice, treatments with or without powder and no choice tests, where insects were confined for five days in a container containing ten grains of lima beans with 0.3 g of powder to be tested. It was observed that some plants have killed the insect while others just repelled the insect, causing no harm to it. Also, one of the studied plants repelled and killed the insects (S. aromaticum, C. ambrosioides and P. tuberculatum). We can conclude that: P. nigrum, P. tuberculatum, S. aromaticum e C. ambrosioides are toxic to the Z. subfasciatus, causing its death. L. nobilis, T. erecta e C. nardus do not present any toxic effect over Z. subfasciatus. C. ambrosioides, S. aromaticum e C. nardus are repellent to Z. subfasciatus. P. nigrum, P. tuberculatum e T. erecta are neutral and M. officinalis have presented no effects over Z. subfasciatus in relation to the parameters evaluated.

Resistance to the weevils Cylas puncticollis and Cylas brunneus conferred by sweetpotato root surface compounds.

Seven resistant varieties of sweetpotato were compared with three susceptible varieties in field trials and laboratory bioassays and showed that resistance was an active process rather than an escape mechanism, as field resistant varieties also had reduced root damage and oviposition compared with susceptible varieties in the laboratory. Liquid chromatography-mass spectrometry (LC-MS) of root surface and epidermal extracts showed significant variation in the concentration of hexadecyl, heptadecyl, octadecyl, and quinic acid esters of caffeic and coumaric acid, with higher concentrations correlated with resistance. All compounds were synthesized to enable their positive identification. Octadecyl coumarate and octadecyl caffeate applied to the surface of susceptible varieties in laboratory bioassays reduced feeding and oviposition, as observed on roots of resistant varieties, and therefore are implicated in weevil resistance. Segregating populations from breeding programs can use these compounds to identify trait loci for resistance and enable the development of resistant varieties.

Freezing as a treatment to prevent the spread of Hypothenemus hampei (Coleoptera: Curculionidae), in coffee.

Coffee berry borer, Hypothenemus hampei (Ferrari) is the most serious insect pest of coffee around the world. Although it is already present in most of the world's major coffee growing regions, it is important to delay further spread and to prevent reintroductions that might include hyperparasites or improve the genetic base of existing populations. Green coffee is shipped around the world for custom blending and roasting and such shipments carry the risk of spreading H. hampei. We used heavily infested coffee berries as a surrogate for green coffee to test the freezing tolerance of H. hampei. After freezing, all life stages of H. hampei were dissected from coffee berries and mortality was assessed. Counting all life stages, > 15,000 insects were measured in this study. A temperature of approximately -15 degrees C (range, -13.9 to -15.5) for 48 h provided 100% control of all life stages. A logit regression model predicted < or = 1 survivor in a million for treatments of -20 degrees C for 5 d or -15 degrees C for 6 d. A freezing treatment for green coffee might be more economical and acceptable compared with fumigation with methyl bromide, especially for small-scale and organic growers and millers in Hawaii who ship green coffee beans to other islands for custom roasting. Freezing treatments could also be used to kill H. hampei in coffee seeds before export with minimal effects on seed germination if coffee seeds are first dried to critical water content levels in accordance with published methods.

Energy storage and C:N:P variation in a holometabolous insect (Curculio davidi Fairmaire) larva across a climate gradient.

Increasing empirical evidence has documented variability in elemental composition within species. However, the extent, causes, and pattern of variability in consumer stoichiometry across a large geographical scale are not well understood. Here, we investigated this issue using a holometabolous insect, weevils (Curculio davidi Fairmaire). Larvae of this species store energy needed for diapause, and variable energy requirements across the geographic range of this species could lead to differences in body elemental composition. Our results showed that variability was high (assessed as the coefficient of variation (CV)) in larval body nitrogen (N) and phosphorus (P) (CV, 10% for N and 13% for P) compared to emerging adults (CV, 5% for N and 8% for P). Temperature-related factors explained more variation than other climatic factors and food for carbon (C), N and P in weevil. In warmer regions, larval C concentration was higher, while N and P were lower. The high C content of weevil larvae relative to both their food source and their adult stage was attributed to energy storage. Across the climatic gradient of its geographic range, larval body C content increased with mean annual temperature and decreased with average diurnal temperature range. This finding implies that temperature-related C storage drives the high variability in elemental composition of larvae across the climate gradient, and also effectively dampens the stoichiometric imbalance between consumers and food resources while serving as an energy reservoir for overwintering and metamorphosis.

Billbug (Coleoptera: Curculionidae) species composition, abundance, seasonal activity, and developmental time in Florida.

Billbugs (Coleoptera: Curculionidae: Sphenophorus spp.) are common pests whose damage is often misdiagnosed on turfgrass in the United States. Consequently, Florida turfgrass managers have been struggling to satisfactorily control billbug outbreaks. Thus, we sought to determine the species complex, abundance, seasonality, and fecundity of key Sphenophorus spp. from field collections, and quantify duration of developmental and daily activity periods through greenhouse rearing tests. From January 2006 to December 2007, > 18,000 adults of 10 different Sphenophorus spp. were collected from four linear pitfall traps on each of two golf courses in north central and two courses in southern Florida. Sphenophorus venatus vestitus Chittenden was the most abundant species, making up > 94% of all specimens collected from three of the four golf courses (80.9% of all specimens collected). Adults were active and mature eggs were present in female ovaries nearly every week of the year. Adults were nocturnal. S. v. vestitus development from egg to adult can occur in 8-9 wk on 'Tifway' bermudagrass [Cynodon dactylon (L.) Pers.] or 'Empire' zoysiagrass (Zoysia japonica Steud.), with up to six overlapping generations per year, depending on environmental conditions. Thus, any time of year could be suitable to target either adults or larvae, and a repeated application may be needed to manage subsequently emerged larvae or eclosed adults.