Repellency of essential oils against Nephotettix cincticeps: Laboratory and glasshouse assays

Four oils from Piper nigrum, Litsea cubeba, Zanthoxylum bungeanum and Curcuma longa were obtained by ethanol extraction. The repellency of these oils and two major compounds (linalool and piperine) was evaluated against female adult and third‐instar nymphs of the rice pest, Nephotettix cincticeps, under laboratory and glasshouse conditions. Paired‐choice and no‐choice assays were used for each treatment, with essential oils evaluated after 24 and 48 hr of exposure and chemical compounds evaluated after 12 and 24 hr of exposure. The potential effects of essential oils on activities of glutathione S tranferase (GST), carboxyl esterase (CarE) and acetyl cholinesterase (AchE) were also evaluated after 48 hr of exposure to leafhoppers. The constituents of the essential oils were determined using GC‐MS. The results showed that the major components in the oils were piperine (34.75%) for P. nigrum, 9,12‐octadecadienoic acid (Z,Z) (18.74%) for L. cubeba, ethanone, 1‐(2‐hydroxy‐4,6‐dimethoxyphenyl) (18.51%) for Z. bungeanum and turmerone (15.89%) for C. longa. In all cases, the essential oils repelled female adults and third‐instar nymphs of N. cincticeps. The repellency of the tested oils and chemicals compounds in the paired‐choice assay was higher than in the no‐choice assay. In all experimental conditions, P. nigrum and C. longa oils were the most and the least potent, respectively. Linalool was the best repellent among the single‐tested compounds under laboratory conditions. In the glasshouse study, the highest repellency was observed in the mixture of linalool and piperine. GST and CarE activities of leafhoppers were significantly enhanced by exposure to the four essentials oils; AchE activity increased significantly only in the P. nigrum and L. cubeba assays. Our results clearly indicate that the tested oils and chemical compounds are promising agents for developing plant‐based pesticides to control N. cincticeps.     

Effectiveness of aromatic plant species for repelling and preventing oviposition of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae)

This study assesses the effectiveness of essential oils for repelling whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), and deterring oviposition. We evaluated essential oils from four plants: Curcuma longa (Zingiberaceae), Litsea cubeba (Lauraceae), Piper nigrum (Piperaceae) and Zanthoxylum bungeanum (Rutaceae), under laboratory and glasshouse conditions. In the laboratory, ethanol‐extracted essential oils were applied uniformly on both sides of a tomato leaf in the treatment groups, and the same volume of liquid but without essential oil was used for the control. Results showed that essential oils from L. cubeba, P. nigrum, C. longa and Z. bungeanum provided some repellency of whiteflies and reduced oviposition. Oils from L. cubeba repelled adult females (69.14% and 62.49% repellency at 24 and 48 hr, respectively), and oils from C. longa deterred oviposition (68.46% and 65.94% at 24 and 48 hr exposures, respectively). In the glasshouse, potted tomato plants with eight leaves were sprayed with treatments and control material. Five‐day‐old adult female whiteflies were released in a cage between both pots. Essential oils from L. cubeba repelled females 54.77 at 24 hr and P. nigrum 44.37% at 48 hr whereas oils from P. nigrum reduced oviposition by 43.35 at 24 hr and Z. bungeanum 21.08% at 48 hr, under glasshouse conditions. Essential oils show potential to be used to reduce feeding and oviposition, and thus being important components of integrated pest management programs. However, future research is needed to investigate individual compounds of these oils against whitefly, and to enhance their effectiveness.     

Transgenic Cry1Ab rice does not impact ecological fitness and predation of a generalist spider

Background

The commercial release of rice genetically engineered to express a Cry1Ab protein from Bacillus thuringiensis (Bt) for control of Lepidoptera in China is a subject of debate. One major point of the debate has focused on the ecological safety of Bt rice on nontarget organisms, especially predators and parasitoids that help control populations of insect pests.

Methodology/Principal Findings

A tritrophic bioassay was conducted to evaluate the potential impact of Cry1Ab-expressing rice on fitness parameters of a predaceous ground spider (Pardosa pseudoannulata (Bösenberg et Strand)) that had fed on Bt rice-fed brown planthopper (Nilaparvata lugens (Stål)) nymphs. Survival, development time and fecundity of this spider were not different when they were fed with Bt rice-fed or non-Bt rice-fed prey. Furthermore, ELISA and PCR gut assays, as well as a functional response trial, indicated that predation by P. pseudoannulata was not significantly different in Bt rice or non-Bt rice fields.

Conclusions/Significance

The transgenic Cry1Ab rice lines tested in this study had no adverse effects on the survival, developmental time and fecundity of P. pseudoannulata in the laboratory or on predation under field conditions. This suggests that this important predator would not be harmed if transgenic Cry1Ab rice were commercialized.     

Nematicidal Activity of Plant Essential Oils and Components From Ajowan (Trachyspermum ammi), Allspice (Pimenta dioica) and Litsea (Litsea cubeba) Essential Oils Against Pine Wood Nematode (Bursaphelenchus Xylophilus)

Commercial plant essential oils from 26 plant species were tested for their nematicidal activities against the pinewood nematode, Bursaphelenchus xylophilus. Good nematicidal activity against B. xylophilus was achieved with essential oils of ajowan (Trachyspermum ammi), allspice (Pimenta dioica) and litsea (Litsea cubeba). Analysis by gas chromatography-mass spectrometry led to identification of 12, 6 and 16 major compounds from ajowan, allspice and litsea oils, respectively. These compounds from three plant essential oils were tested individually for their nematicidal activities against the pinewood nematode. LC₅₀ values of geranial, isoeugenol, methyl isoeugenol, eugenol, methyl eugenol and neral against pine wood nematodes were 0.120, 0.200, 0.210, 0.480, 0.517 and 0.525 mg/ml, respectively. The essential oils described herein merit further study as potential nematicides against the pinewood nematode.     

Effects of Volatiles of Essential Oils on Behavior of the Western Flower Thrips <Emphasis Type="Italic">Frankliniella occidentalis</Emphasis> Perg. (Thysanoptera,Thripidae)

The results of evaluating the effects of essential oils on the behavior and reproduction potential of the western flower thrips Frankliniella occidentalis Perg. are presented. Essential oils of Acorus calamus, Juniperus virginiana, and Melissa officinalis possessed a repellent effect on the larvae. A sample from Lutsea cubeba was characterized by an attractive property. Oil solutions of J. virginiana, Mentha spicata, Nepeta cataria, and Litsea cubeba repelled the thrips females off the leaves of the host plant. Reduced concentrations of oils from J. virginiana, M. officinalis, and M. spicata did not affect the distribution of thrips females between the experimental and control leaves, but the number of offspring was significantly lower on the treated leaves.     

Chemical Composition and Antibacterial Activity of Essential Oils from Different Parts of Litsea cubeba

The composition of the essential oils obtained by hydrodistillation of different parts of Litsea cubeba, including roots, stems, leaves, alabastra (flower buds), flowers, and fruits, were investigated by GC (RI) and GC/MS. The antimicrobial activity of the oils was assessed with disc diffusion and microbroth dilution assays. The results showed large variations in the composition among the different oils. The major components in the oils from roots and fruits, from stems, leaves, and alabastra, and from flowers were citral B (neral), β‐phellandrene, and β‐terpinene, respectively. The inhibition zone (DD) and MIC values for the bacterial strains tested, which were all sensitive to the essential oil of L. cubeba, were in the range of 10.1–35.0 mm and 100–1000 μg/ml, respectively. Hence, the oils of the various parts showed moderate activity against the tested bacteria. This investigation showed that the antibacterial activity of L. cubeba was attributed to the essential oils, thus they can be a potential medicinal resource.     

Anti-biofilm activity of LC-MS based Solanum nigrum essential oils against multi drug resistant biofilm forming P. mirabilis

Urinary tract infections are second most important diseases worldwide due to the increased amount of antibiotic resistant microbes. Among the Gram negative bacteria, P. mirabilis is the dominant biofilm producer in urinary tract infections next to E. coli. Biofilm is a process that produced self-matrix of more virulence pathogens on colloidal surfaces. Based on the above fact, this study was concentrated to inhibit the P. mirabilis biofilm formation by various in-vitro experiments. In the current study, the anti-biofilm effect of essential oils was recovered from the medicinal plant of Solanum nigrum, and confirmed the available essential oils by liquid chromatography-mass spectroscopy analysis. The excellent anti-microbial activity and minimum biofilm inhibition concentration of the essential oils against P. mirabilis was indicated at 200 µg/mL. The absence of viability and altered exopolysaccharide structure of treated cells were showed by biofilm metabolic assay and phenol–sulphuric acid method. The fluorescence differentiation of P. mirabilis treated cells was showed with more damages by confocal laser scanning electron microscope. Further, more morphological changes of essential oils treated cells were differentiated from normal cells by scanning electron microscope. Altogether, the results were reported that the S. nigrum essential oils have anti-biofilm ability.     

Synergistic repellent and irritant effect of combined essential oils on Aedes aegypti (L.) mosquitoes

This study was designed to compare the behavioral responses of Aedes aegypti to a single essential oil and to a mixture of two or three essential oils using an excito‐repellency test chamber. Mixtures were prepared from essential oils extracted from Litsea cubeba (LC), Litsea salicifolia (LS), and Melaleuca leucadendron (ML). In general, the mixture of essential oils produced a much stronger escape response by Ae. aegypti, regardless of the test conditions. No significant difference in escape responses was seen when the mixture of oils was compared with a standard commercial product containing DEET. Greater contact irritancy was seen from mixed oils of LC and LS than with other mixed oils. Mixtures of LC and LS at 0.075% showed the highest synergistic action (65.5% escaped) compared to that with unmixed oil alone at the same concentration (LC/20% and LS=32.2%). In addition, mixtures of LC and LS at 0.075% demonstrated the highest non‐contact repellency (62.7%) and showed a greater effect than the use of LC (20%) or LS (20.3%) alone. We conclude that mixtures of two essential oils show potential as active ingredients for mosquito repellents.     

Lignan profile of Piper cubeba, an Indonesian medicinal plant

The lignan profile of the aerial part of Piper cubeba L. (Piperaceae) was determined using GC, GC–MS and HPLC. The number of lignans found in the leaves was 15, followed by berries and the stalks with respectively 13 and five lignans. This is the first investigation of lignans from the leaves and the stalks of P. cubeba. Cubebin, hinokinin, yatein, isoyatein are common lignans in the genus Piper and appeared as major components in all parts of P. cubeba investigated.     

低剂量杀虫剂对星豹蛛捕食效应的影响及其机理

探讨低剂量杀虫剂对蜘蛛捕食效应的影响及其生化机理。采用药膜法,测定了低剂量吡虫啉作用下,星豹蛛和甘蓝蚜敏感性、捕食效应以及成蛛体内乙酰胆碱酯酶(AChE)、谷胱甘肽S-转移酶(GSTs)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和中肠蛋白消化酶的活力变化。低剂量农药作用下,星豹蛛对甘蓝蚜的功能反应类型为HollingⅡ型,与对照组相比,随着猎物密度的增大星豹蛛的捕食量增加,寻找效应降低,对猎物的处理时间Th缩短,从而增强了对猎物的捕食作用;低剂量农药处理后,星豹蛛体内AChE和GSTs活性均显著低于对照组(P0.05),说明酶活性受到抑制,且抑制作用随吡虫啉浓度的增大而加强,随作用时间的延长而减弱;SOD,CAT和中肠蛋白消化酶活性显著增强,与对照组相比存在显著或极显著差异(P0.05),且随吡虫啉浓度增大和作用时间的延长而逐渐降低,最后接近对照组。低剂量杀虫剂作用下,蜘蛛体内AChE活性受到抑制,AChE敏感性降低,对神经递质乙酰胆碱的分解作用下降,使蜘蛛的兴奋性增加;GSTs、SOD、CAT等代谢酶活性发生变化,使蜘蛛的新陈代谢加速,从而刺激捕食;中肠蛋白消化酶的活性增强,提高了对猎物的消化吸收功能。总之,在低剂量杀虫剂作用下,星豹蛛通过外在的捕食行为和体内一系列酶系生理生化反应的综合作用促进蜘蛛对害虫的控制作用。     

Insecticidal activity of five Chinese medicinal plants against Plutella xylostella L. larvae

Twenty-five extracts of five Chinese medicinal plant species were screened for insecticidal activity against the diamondback moth, Plutella xylostella L., larvae by a leaf dipping bioassay method. The ethyl acetate, ethanol, and acetone extracts of Veratrum nigrum L. root and rhizome, the ethyl acetate extract of Phytolacca americana L. root, and the petroleum ether extract of Pseudolarix kaempferi Gord. root bark were effective against P. xylostella. Among them, the ethyl acetate extract of V. nigrum showed the strongest insecticidal activity against the second and third instar larvae of P. xylostella, with LC₅₀ values of 225 and 335 ppm, respectively. The other extracts gave little or no insecticidal activity against P. xylostella. These results indicated that V. nigrum may be a promising naturally occurring agent for P. xylostella larval control, and that the organic solvent used for plant extraction can affect its insecticidal activity.     

Next-generation sequencing analysis of Pardosa pseudoannulata&#x27;s diet composition in different habitats

Spiders are the most common and predominant predators in terrestrial ecosystems. The predatory behavior of spiders affects the energy flow across the food web within an ecosystem. Traditiaonal methods for analyzing spider diets such as field observation, anatomy and faeces analysis are not suitable for spider experiments due to spiders’ special dietary behavior. The molecular method based on the specific primers of prey DNA seems to be inefficient either in spite of its wide application in diet analysis. As the next-generation sequencing (NGS) technology becomes prevalent in many different areas, several cases of the NGS-based analysis of mammal diets have been published. This study analyzed the diet differences of Pardosa pseudoannulata (Araneae: Lycosidae) in four habitats (a wetland, a tea plantation, an alpine meadow and a paddy field) by using the NGS technology, combined with the DNA barcode method. The results suggested that the Pardosa pseudoannulata feed on a broad range of prey, and 7 orders and 24 families of insects were detected in the four investigated habitats. Moreover, it is found that the diet diversity of Pardosa pseudoannulata is greatly influenced by their living environments and seasons. In a nutshell, this study established an NGS-based methodology for spider diets analysis, and the results provided some basic materials to inform the protection and utilization of the Pardosa pseudoannulata as a potential eco-friendly predator against pests.     

Biological control: An effective approach against nematodes using black pepper plants (Piper nigrum L.)

Black pepper (Piper nigrum L.) is one of the oldest spices in the world, additionally, it is highly demanded. Several biotic and abiotic variables pose black pepper production worldwide. Plant-parasitic nematodes play a key role among biotic factors, causing considerable economic losses and affecting the production. Different synthetic nematicides were used for controlling plant nematodes, however the majority of pesticides have been pulled from the market due to substantial non-target effects and environmental risks. As a result, the search for alternative eco-friendly agents for controlling plant-parasitic nematodes populations. Microbial agents are a precious option. In this review the bacterial and fungal agents used as an alternative nematicides, they were studied and confirmed as essential anti-microbial agents against plant nematodes which infected Piper nigrum L. This work examines the most common plant nematodes infected Piper nigrum L., with a focus on root knot and burrowing nematodes, in addition, how to control plant parasitic nematodes using microorganisms.     

Effect of Cry1Ab protein on hemocytes of the wolf spider Pardosa pseudoannulata

Studies on the effect of Cry1Ab protein on hemocytes of the wolf spider Pardosa pseudoannulata revealed that Cry1Ab protein could accumulate in the four-instar and adult spiders via the food chain from transgenic rice expressing Cry1Ab protein through its prey brown planthoppers with approximate 20-time enrichment, but could not accumulate in hemolymph of the spider. The accumulated Cry1Ab had no significant effects on several elements of hemolymph including stored energy, calcium ion concentration and apoptosis rate of the hemocytes, indicating that Cry1Ab could not affect the hemocytes of P. pseudoannulata.     

The effects of laurel (Laurus nobilis L.) on development of two mycorrhizal fungi

Hundreds of aromatic plant species are growing naturally around Mediterranean. Plant essential oils are incorporated in aromatic plant material and follow the litter fall. During litter degradation, the presence of essential oils can affect soil microorganisms. Mycorrhizal fungi have never been investigated so far under the presence of volatile oils. The aim of this study was to explore the effect of aromatic Laurus nobilis L. on development of two mycorrhizal species Glomus deserticola and Glomus intraradices. The response of fungi colonization and host growth were monitored under different concentrations of L. nobilis leaves and essential oil. The major compounds of L. nobilis essential oil were 1,8-cineole (49.6%), sabinene (7.8%), ??-pinene (6.0%), eugenole (5.6%), ??-terpinyl acetate (5.2%) and ??-pinene (5.1%). Both mycorrhizal fungi colonized successfully the host plants whose growth was positively influenced by mycorrhizal fungi. G. deserticola presented higher infection level than G. intraradices. The addition of L. nobilis leaves in the soil resulted in mycorrhiza inhibition. The level of inhibition was positively correlated with the added amount of aromatic leaves in the soil. The essential oil presented a little higher inhibition than the leaves. The presence of this aromatic plant in many different ecosystems could contribute in mycorrhiza inhibition and it is suggested, when it’s possible, reduction of laurel litter before reforestation programs.     

Identification of two acetylcholinesterases in Pardosa pseudoannulata and the sensitivity to insecticides

Pardosa pseudoannulata is an important predatory enemy against insect pests, such as rice planthoppers and leafhoppers. In order to understand the insecticide selectivity between P. pseudoannulata and insect pests, two acetylcholinesterase genes, Pp-ace1 and Pp-ace2, were cloned from this natural enemy. The putative proteins encoded by Pp-ace1 and Pp-ace2 showed high similarities to insect AChE1 (63% to Liposcelis entomophila AChE1) and AChE2 (36% to Culex quinquefasciatus AChE2) with specific functional motifs, which indicated that two genes might encode AChE1 and AChE2 proteins respectively. The recombinant proteins by expressing Pp-ace1 and Pp-ace2 genes in insect sf9 cells showed high AChE activities. The kinetic parameters, V_max and K_m, of two recombinant AChE proteins were significantly different. The sensitivities to six insecticides were determined in two recombinant AChEs. Pp-AChE1 was more sensitive to all tested insecticides than Pp-AChE2, such as fenobucarb (54 times in K_i ratios), isoprocarb (31 times), carbaryl (13 times) and omethoate (6 times). These results indicated that Pp-AChE1 might be the major synaptic enzyme in the spider. By sequence comparison of P. pseudoannulata and insect AChEs, the key amino acid differences at or close to the functional sites were found. The locations of some key amino acid differences were consistent with the point mutation sites in insect AChEs that were associated with insecticide resistance, such as Phe331 in Pp-AChE2 corresponding to Ser331Phe mutation in Myzus persicae and Aphis gossypii AChE2, which might play important roles in insecticide selectivity between P. pseudoannulata and insect pests. Of course, the direct evidences are needed through further studies.     

Effects of vanillin, quillaja saponin, and essential oils on in vitro fermentation and protein-degrading microorganisms of the rumen

This study investigated the effects of vanillin on methanogenesis and rumen fermentation, and the responses of ruminal protein-degrading bacteria to vanillin (at concentrations of 0, 0.76 and 1.52 g/L), essential oils (clove oil, 1 g/L; origanum oil, 0.50 g/L, and peppermint oil, 1 g/L), and quillaja saponin (at concentration of 0 and 6 g/L) in vitro. Methane production, degradabilities of feed substrate, and ammonia concentration decreased linearly with increasing doses of vanillin. Concentration of total volatile fatty acids also decreased, whereas proportion of butyrate tended to increase linearly with increasing doses of vanillin. Protozoa population decreased, but abundances of Ruminococcus flavefaciens, Prevotella bryantii, Butyrivibrio fibrisolvens, Prevotella ruminicola, Clostridium aminophilum, and Ruminobacter amylophilus increased with increasing doses of vanillin. Origanum and clove oils resulted in lower ammonia concentrations compared to control and peppermint oil. All the tested essential oils decreased abundances of protozoa, Selenomonas ruminantium, R. amylophilus, P. ruminicola and P. bryantii, with the largest decrease resulted from origanum oil followed by clove oil and peppermint oil. The abundances of Megasphaera elsdenii, C. aminophilum, and Clostridium sticklandii were deceased by origanum oil while that of B. fibrisolvens was lowered by both origanum and clove oils. Saponin decreased ammonia concentration and protozoal population, but increased the abundances of S. ruminantium, R. amylophilus, P. ruminicola, and P. bryantii, though the magnitude was small (less than one log unit). The results suggest that reduction of ammonia production by vanillin and saponin may not be caused by direct inhibition of major known proteolytic bacteria, and essential oils can have different inhibitory effects on different proteolytic bacteria, resulting in varying reduction in ammonia production.     

Toxicity,repellent and oviposition deterrent effects of select essential oils against the house fly Musca domestica

Essential oils from plants may provide environment-friendly alternatives to conventional synthetic insecticides. Here, toxic, repellent, and oviposition deterrent effects of essential oils of six plants: Allium sativum L. (Alliaceae), Azadirachta indica A. Juss. (Meliaceae), Cinnamomum cassia (L.) (Lauraceae), Eucalyptus camaldulensis Dehnh. (Myrtaceae), Piper nigrum L. (Piperaceae), and Thevetia peruviana (Pers.) (Apocynaceae), were evaluated against different life stages of Musca domestica. Bioassays revealed that the essential oils of A. indica, T. peruviana and E. camaldulensis exhibited: a) the highest toxicity on larvae (LC₅₀ = 169.72, 182.23 and 277.01 ppm, respectively), pupae (LC₅₀ = 150.56, 164.84 and 164.87 ppm, respectively) and adults (LC₅₀ = 166.69, 139.15 and 302.75 ppm, respectively) of M. domestica; b) the highest repellency (91.44, 72.19 and 72.80%, respectively) and oviposition deterrent (90.36, 88.82 and 89.13%, respectively) effects on adults of M. domestica, as compared to the other essential oils. Moreover, the speed of mortality caused by essential oils of A. indica (LT₅₀ = 16.85 and 17.06 h for larvae and adults, respectively) and T. peruviana (LT₅₀ = 16.46 and 18.58 h for larvae and adults, respectively) was faster than the rest of the essential oils. On the whole, it might be expected that the essential oils of A. indica, T. peruviana and E. camaldulensis could be developed into a new type of environment-friendly insecticides and/or repellents for the management of M. domestica.