Neuroprotective and Anti-Amyloid ß Effect and Main Chemical Profiles of White Tea: Comparison Against Green, Oolong and Black Tea.

White tea (WT) is one of six tea types originally derived from Fujian Province, China. White tea is known for its health-promoting properties. However, the neuroprotective and anti-aggregatory properties of WT against the hallmark toxic Alzheimer's protein, Aß have not been investigated. In this study, WT, green tea (GT), oolong tea (OT) and black tea (BT) were manufactured using tea leaves from the cultivar Camellia sinensis (Jin Guanyin). The protective effects of these tea extracts were then studied under oxidative stress conditions via t-bhp and H2O2 exposure, in addition to Aß treatment using a PC-12 cell model. Each tea type failed to rescue PC-12 cells from either t-bhp or H2O2-mediated toxicity, however each extract exerted significant protection against Aß-evoked neurotoxicity. Results of the Thioflavin T Kinetic (ThT) and TEM assay showed that Aß aggregate formation was inhibited by each tea type. Additionally, TEM also supported the different anti-aggregatory effect of WT by modifying Aß into an amorphous and punctate aggregate morphology. Higher accumulated precedent or potential neuroprotective compounds in WT, including ECG''3Me, 8-C-ascorbyl-EGCG, GABA and Gln, in addition to flavonol or flavone glycosides detected by using UPLC-QTOF-MS and UPLC-QqQ-MS, may contribute to a favourable anti-aggregative and neuroprotective effect of WT against Aß.

Choice and No-Choice Bioassays to Study the Pupation Preference and Emergence Success of Ectropis grisescens.

Many insects live above the ground as larvae and adults and as pupate below the ground. Compared to the above-ground stages of their life cycles, less attention has been paid on how environmental factors affect these insects when they pupate within the soil. The tea looper, Ectropis grisescens Warren (Lepidoptera: Geometridae), is a severe pest of tea plants and has caused huge economic losses in South China. The protocols described here aim to investigate, through multiple-choice bioassays, whether mature last-instar E. grisescens larvae can discriminate soil variables such as the substrate type and moisture content, and determine, through no-choice bioassays, the impact of the substrate type and moisture content on pupation behaviors and the emergence success of E. grisescens. The results would enhance the understanding of the pupation ecology of E. grisescens and may bring insights into soil-management tactics for suppressing E. grisescens populations. In addition, these bioassays can be modified to study the influences of various factors on the pupation behaviors and survivorship of soil-pupating pests.

Reference genes selection for quantitative gene expression studies in tea green leafhoppers, Empoasca onukii Matsuda.

Empoasca onukii Matsuda is one of the most devastating pests of the tea plant (Camellia sinensis). Still, the presumed expression stability of its reference genes (RGs) has not been analyzed. RGs are essential for accurate and reliable gene expression analysis, so this absence has hampered the study of the insect's molecular biology. To find candidate RGs for normalizing gene expression data, we cloned ten common housekeeping genes from E. onukii. Using the ΔCt method, geNorm, NormFinder and BestKeeper, we screened the RGs that were appropriate for quantifying the mRNA transcription of cellular responses under five experimental conditions. We identified the combinations of α-TUB and G6PDH, α-TUB and UBC, two RGs (α-TUB and ß-TUB1) or three RGs (α-TUB, RPL13 and GAPDH), AK and UBC, or RPL13 and α-TUB as the best for analyzing gene expression in E. onukii adults of both sexes in different tissues, nymphs at different developmental stages, nymphs exposed to different temperatures or nymphs exposed to photoperiod stress. Finally, the E. onukii cysteine proteinase (Eocyp) was chosen as the target gene to validate the rationality of the proposed RGs. In conclusion, our study suggests a series of RGs with which to study the gene expression profiles of E. onukii that have been manipulated (biotically or abiotically) using reverse transcription quantitative polymerase chain reaction. The results offer a solid foundation for further studies of the molecular biology of E. onukii.

Tea silkworm droppings as an enriched source of tea flavonoids.

Andraca droppings is the waste excreted from the tea biter Andraca theae. Its chemical constituents and potential medical use, unlike those of the traditional Chinese medicine silkworm droppings, have not been reported yet. To explore new nutraceuticals, the chemical constituents of this substance were investigated. Since the bioactive ingredients are generally present in the EtOAc-soluble fraction, this fraction, obtained from the ethanolic extract of the dried Andraca droppings by liquid-liquid partitioning, was separated by chromatographic methods, including Sephadex LH-20, centrifugal partition chromatography, and RP-18 columns, to produce 14 compounds (1-14). They were characterized as 1,7-dimethyl xanthine (1), three benzoic acids (2, 3, and 5), and 10 flavonoids (4, 6-14). The amount of compounds 6, 7, 10, 13, and 14 in the droppings were 1.7-15.5-fold compared to those of tea leaves. In addition, 1,7-dimethyl xanthine (1) was found present only in the Andraca droppings but absent in tea leaves. Therefore, except for compound 1, which might be transformed from caffeine by microflora in the insect, the compounds were believed not to be absorbed by the worm gut and excreted directly. The present study suggests the Andraca droppings are an enriched source of the bioactive flavonoids from tea leaves and are potential as a useful nutraceutical.

Molecular characterization of tea mosquito bug from tea growing regions of India.

The tea mosquito bug, Helopeltis (Hemiptera: Miridae), is an insidious pest that poses a significant economical threat to tea plantations. As a basic first step to control this pest is authentic identification, but the inability to determine morphological characters of Helopeltis species makes this process very difficult. DNA barcoding is a reliable alternative to traditional morphological identification of this pest. Since tea is cultivated in different parts of the country, an attempt was made to molecular characterization of Helopeltis. This is the first report on molecular identification and diversity characterization of Helopeltis collected from tea growing regions of southern and north India, using cytochrome c oxidase subunit I (COI) gene of mitochondrial (mt) DNA. Beginning with the molecular identification of this pest is essential to start an effective pest management strategy, and will provide basic information for diffusion pattern, population dynamics and chemical application.

Field efficacy and transmission of fast- and slow-killing nucleopolyhedroviruses that are infectious to Adoxophyes honmai (Lepidoptera: Tortricidae).

The smaller tea tortrix, Adoxophyes honmai (Lepidoptera: Tortricidae), is an economically important pest of tea in Japan. Previous work showed that a fast-killing nucleopolyhedrovirus (NPV) isolated from A. orana (AdorNPV) and a slow-killing NPV isolated from A. honmai (AdhoNPV) are both infectious to A. honmai larvae. Field application of these different NPVs was conducted against an A. honmai larval population in tea plants, and the control efficacy and transmission rate of the two NPVs were compared. The slow-killing AdhoNPV showed lower field efficacy, in terms of preventing damage caused by A. honmai larvae against the tea plants, than the fast-killing AdorNPV. However, AdhoNPV had a significantly higher horizontal transmission rate than AdorNPV. These results show that AdorNPV is suitable as an inundative agent, while AdhoNPV is an appropriate inoculative agent.

Volatiles emitted from tea plants infested by Ectropis obliqua larvae are attractive to conspecific moths.

Herbivore-induced plant volatiles have been reported to play a role in the host-searching behavior of herbivores. However, next to nothing is known about the effect of volatiles emitted from tea plants infested by Ectropis obliqua larvae on the behavior of conspecific adults. Here, we found that tea plants infested by E. obliqua caterpillars for 24 h were more attractive to both virgin male and female E. obliqua adults than were intact, uninfested tea plants; moreover, mated female E. obliqua moths were more attracted by infested tea plants and preferentially oviposited on these plants, whereas male moths were repelled by infested plants once they had mated. Volatile analysis revealed that the herbivore infestation dramatically increased the emission of volatiles. Among these volatiles, 17 compounds elicited antennal responses from both male and female virginal moths. Using a Y-tube olfactometer, we found that 3 of the 17 chemicals, benzyl alcohol, (Z)-3-hexenyl hexanoate, and (Z)-3-hexenal, were attractive, but two compounds, linalool and benzyl nitril, were repellent to virgin male and female moths. One chemical, (Z)-3-hexenyl acetate, was attractive only to virgin males. Mated females were attracted by three compounds, (Z)-3-hexenyl hexanoate, (Z)-3-hexenyl acetate, and (Z)-3-hexenal; whereas mated males were repelled by (Z)-3-hexenol. The findings provide new insights into the interaction between tea plants and the herbivores, and may help scientists develop new measures with which to control E. obliqua.

Enhanced conidial thermotolerance and field efficacy of Beauveria bassiana by saccharides.

Insecticide efficacy of Beauveria bassiana conidia was improved by optimizing the concentrations of conidial heat-protective saccharides (glucose, sucrose, maltose, trehalose, α-lactose, and mannitol) using response surface methodology. Two field trials in tea gardens were carried out to control leafhopper (Empoasca vitis) by spraying B. bassiana conidia together with the optimized saccharides (0.26 g glucose, 0.28 g lactose, 0.24 g mannitol per ml). In the field studies, B. bassiana conidia were applied to control Empoasca vitis with and without saccharides and compared with bifenthrin, a pyrethroid insecticide. With the optimal concentrations of saccharides, the conidial germination rate reached 72% and the control efficacy of the saccharides group (65.7%) was equal to the bifenthrin group (69.4%), which improved by about 55%.

Detecting deep divergence in seventeen populations of tea geometrid (Ectropis obliqua Prout) in China by COI mtDNA and cross-breeding.

The tea geometrid (Ectropis obliqua Prout, Lepidoptera: Geometridae) is a dominant chewing insect endemic in most tea-growing areas in China. Recently some E. obliqua populations have been found to be resistant to the nucleopolyhedrovirus (EoNPV), a host-specific virus that has so far been found only in E. obliqua. Although the resistant populations are morphologically indistinguishable from susceptible populations, we conducted a nationwide collection and examined the genetic divergence in the COI region of the mtDNA in E. obliqua. Phylogenetic analyses of mtDNA in 17 populations revealed two divergent clades with genetic distance greater than 3.7% between clades and less than 0.7% within clades. Therefore, we suggest that E. obliqua falls into two distinct groups. Further inheritance analyses using reciprocal single-pair mating showed an abnormal F1 generation with an unbalanced sex ratio and the inability to produce fertile eggs (or any eggs) through F1 self-crossing. These data revealed a potential cryptic species complex with deep divergence and reproductive isolation within E. obliqua. Uneven distribution of the groups suggests a possible geographic effect on the divergence. Future investigations will be conducted to examine whether EoNPV selection or other factors prompted the evolution of resistance.

Catechol--an oviposition stimulant for cigarette beetle in roasted coffee beans.

The cigarette beetle, Lasioderma serricorne, is a serious global pest that preys on stored food products. Larvae of the beetle cannot grow on roasted coffee beans or dried black or green tea leaves, although they oviposit on such products. We investigated oviposition by the beetles on MeOH extracts of the above products. The number of eggs laid increased with an increase in dose of each extract, indicating that chemical factors stimulate oviposition by the beetles. This was especially true for \ coffee bean extracts, which elicited high numbers of eggs even at a low dose (0.1 g bean equivalent/ml) compared to other extracts. Coffee beans were extracted in hexane, chloroform, 1-butanol, MeOH, and 20% MeOH in water. The number of eggs laid was higher on filter papers treated with chloroform, 1-butanol, MeOH, and 20% MeOH in water extracts than on control (solvent alone) papers. The chloroform extract was fractionated by silica-gel column chromatography. Nine compounds were identified by gas chromatography/mass spectrometry from an active fraction. Of these compounds, only a significant ovipositional response to catechol was observed.

Aerodynamic advantages of upside down take-off for aerial dispersal in Tetranychus spider mites.

Aerial dispersal may be important for redistribution of spider mites into new habitats. Evidence for behavioral control of aerial take-off has been well documented for Tetranychus urticae Koch. Before aerial dispersal they exhibit the aerial take-off posture that involves lifting the forelegs upright and raising the forebody. However, whether the aerial take-off posture functions to increase drag has remained unclear. The objectives of this study were to clarify: (i) aerodynamic effects of the aerial take-off posture; and (ii) actual aerial take-off behavior in T. urticae. To evaluate the aerodynamic forces experienced by grounded spider mites in different postures, we constructed three-dimensional models of T. urticae, exhibiting the aerial take-off posture and the normal posture, using computer graphics. We found that the aerial take-off posture was effective in receiving greater rearward forces from wind rather than upward forces. As a result, aerial take-off from a horizontal platform is unlikely. Instead, inverted departure surfaces, e.g., lower leaf surfaces, with inclines are likely to be effective sites for take-off. Laboratory experiments and field observations indicated that the mites preferentially adopted such a position for orientation and take-off. Our findings provided a rationale for the take-off behavior of Tetranychus spider mites.

Genome sequence and organization of a nucleopolyhedrovirus that infects the tea looper caterpillar, Ectropis obliqua.

The complete nucleotide sequence of Ectropis obliqua nucleopolyhedrovirus (EcobNPV), which infects the tea looper caterpillar, was determined and analyzed. The double stranded circular genome is composed of 131,204 bp and is 37.6% G+C rich. The analysis predicted 126 putative, minimally overlapping open reading frames (ORFs) with 150 or more nucleotides that together compose 89.8% of the genome. The remaining 10.2% constitute non-coding and three homologous regions. Comparison with previously sequenced baculoviruses indicated that three ORFs were unique to EcobNPV, while the remaining 123 ORFs shared identity with other baculovirus genes. In addition to two bro homologues, three other repeat ORFs, including dbp, p26, and odv-e66, were identified. Phylogenetic analysis indicated that each member of the paired ORFs was acquired independently. Gene parity plot analysis and percent identity of gene homologues suggested that EcobNPV is a Group II NPV, although its genomic organization was highly distinct.

[Aromatic constituents in fresh leaves of Lingtou Dancong tea under Basilepta melanopus Lefevre stress].

The study showed that under different degrees of Basilepta melanopus Lefevre stress, the kinds of aromatic constituents in the fresh leaves of Lingtou Dancong tea plant increased evidently, being the most (36 kinds) under medium stress. With the increase of the stress, the contents of 9 aromatic constituents like 1,6-methylene annulene increased, whereas those of 8 constituents as linalool decreased. Light stress induced the appearance of 4 aromatic constituents like 2-methyl hexadecane, while medium stress induced that of 6 constituents as H1-docosanol. Different degrees of B. melanopus stress could induce the synthesis of 23 aromatic constituents like 1,6-methylene annulene, but inhibit the formation of 6 constituents as nonanal.

[Efficacy of emulsifiable formulations of two entomopathogenic fungi against small green leafhoppers on tea plant].

A field trial was conducted to test the efficacy of emulsifiable formulations of Beauveria bassiana (Bb) and Paecilomyces fumosoroseus (Pf) conidia in controlling small green leafhoppers (Empoasca spp.) on tea plant in a hillside tea garden located in Shuichang, Zhejiang during mid-summer 2002. Both Bb and Pf formulations contained 10(10) conidia x ml(-1). Adding 3% of imidacloprid 10% WP to each fungal formulation (W/V) resulted in two other formulations, i.e., Bb-imidacloprid mixture (BbIM) and Pf-imidacloprid mixture (PfIM). Besides the four formulations, a mineral oil-based liquid used to formulate the fungal conidia and containing 3% of imidacloprid 10% WP (OBLI) was also tested, and water spray was used as control (CK). Each of the treatments included three 60-m2 plots (replicates), which were sprayed twice with a 500-fold aqueous dilution of the corresponding formulation or preparation at a 12-d interval. Based on the leafhopper densities estimated by sampling in all plots at 3- or 4-d intervals after the first spray, the spray of BbIM or PfIM could better control the leafhoppers than the spray of Bb or Pf formulation. The maximal efficacy relative to CK reached 83.4% and 71.3% in the BbIM and PfIM treatments, respectively. Estimates of field efficacy obtained during the 25-d period after the first spray was 66.8% for BbIM, 62.1% for PfIM, 50.3% for OBLI, 49.5% for Bb, and 19.0% for Pf, respectively. A discussion was also given on the effect of local weather pattern and leafhopper population age structure on the results of the field trial, and on the application techniques to enhance the efficacy of mycoinsecticides against Empoasca species on tea plant.

Sequence analysis and genomic organization of a new insect picorna-like virus, Ectropis obliqua picorna-like virus, isolated from Ectropis obliqua.

The complete nucleotide sequence of a new insect picorna-like virus, Ectropis obliqua picorna-like virus (EoPV), which causes a fatal infection of Ectropis obliqua larvae, has been determined. The genomic RNA of EoPV is 9394 nt in length and contains a single, large open reading frame (nt 391-9351) encoding a polyprotein of 2987 aa. Sequence comparisons with other viral polyproteins revealed that the consensus sequences for picornavirus RNA helicase, protease and RNA-dependent RNA polymerase proteins are found on the genome in order in the 5'-->3' direction. All structural genes were located at the 5' terminus. In terms of sequence similarity, identity and genome organization, EoPV resembles mammalian picornaviruses and three other insect picorna-like viruses: Infectious flacherie virus of silkworm, Sacbrood virus of honeybee and Perina nuda picorna-like virus (PnPV). Phylogenetic analysis showed that EoPV is most closely related to PnPV and suggests that these four insect picorna-like viruses might constitute a new group of insect-infectious RNA viruses.

Composition of the volatiles from intact and mechanically pierced tea aphid-tea shoot complexes and their attraction to natural enemies of the tea aphid.

The volatile components from intact tea shoots (ITS), obtained by air entrainment, were identified by their mass spectra and retention times and confirmed by comparison with standard samples. They are E-2-hexenal, ocimene, Z-3-hexenyl acetate, Z-3-hexen-1-ol, butanoic acid-3-hexenyl ester, linalool, 1-octanol, geraniol, and indole. Volatiles from mechanically pierced tea shoots (MPTS) were identified as E-2-hexenal, ocimene, Z-3-hexen-1-ol, butanoic acid-3-hexenyl ester, linalool, geraniol, indole, E-2-hexenoic acid, Z-3-hexenyl formate, methyl salicylate, and benzyl alcohol, and volatiles from tea aphid-tea shoot complexes (TATSC) were identified as E-2-hexenal, ocimene, Z-3-hexenyl acetate, Z-3-hexen-1-ol, linalool, geraniol, indole, benzaldehyde, and E-2-hexenoic acid. Z-3-Hexen-1-ol is the main component in the three different types of volatiles, and the amount of benzaldehyde in TATSC volatiles is very ample. The attraction of the volatiles from ITS, MPTS, and TATSC, and the nine components of TATSC volatiles to the natural enemies, the coccinellid, Coccinella septempunctata, the parasite, Aphidius sp., and the lacewing, Chrysopa sinica, were determined by electroantennogram (EAG) and the wind tunnel bioassay. TATSC volatiles and benzaldehyde elicited much larger EAG responses and stronger upwind flight and arresting behavior from each natural enemy in the wind tunnel than other infochemicals.

Transmission of bancroftian filariasis in tea agro-ecosystem of Assam, India.

Tea industry is a labor intensive agro-industry and filariasis is mostly localized among the tea garden workers in Assam. The workers live inside the gardens in colonies. Studies conducted in two cosecutive years revealed that among the host seeking Culex quinquefasciatus average infection rate was 4.6% and with 2.1 larvae per mosquito. The overall prevalence of infective mosquitos was 0.8% with average L3 load of 2.0 per mosquito. The probability of infected mosquitos surviving to have complete development of filarial larvae (13 days) was 0.17. The expectation of infective life was 1.416 days for man biting Cx. quinquefasciatus and the estimated adult survival rate of was 87.6%. It has been estimated that a total of 22,569 mosquito bites were received/man/year in tea garden environment out of which 182 bites/man/year were infective (0.806%). The monthly biting rate varied from 310-4,758.5 bites per man (mean 1,846 +/- 1,389.7 SD). Monthly transmission index of W. bancrofti filaria showed two periods of transmission. In both the year no infection was detected during February and March and infection rate remained low up to May (average infection in April 0.72% and in May 0.48%).

[Behavior response of four Leis axyridis varieties to volatiles from tea and Toxoptera aurantii].

The behavior response of the adults of four Leis axyridis varieties to kairomone from Taxoptera aurantii was determined by Y-shape olfaction instrument. The results showed that when supplied with two-two-combinations of Taxoptera aurantii, Brevicoryne brassicae and Lipaphis erysimi, tea aphids plus aphid-damaged vs. normal shoots, and aphid-damaged vs. normal shoots, Leis axyridis had a preference for tea aphids, tea aphids plus aphid-damaged tea shoots, or aphid-damaged tea shoots. There existed a Logistic curve relationship(P < 0.01) between odor source tendency rate of Leis axyridis and odor source quality, when 8-9 kinds of odor gradients including tea aphids plus aphid-damaged tea shoots, aphid-damaged tea shoots and tea aphids were used as odor sources. The tendency rates of Leis axyridis ab. bimaculata, Leis axyridis ab. conspicua and Leis axyridis var. novemdecimpunctata showed single left-tilting peak curves to the aphid number(P < 0.01). Ether and n-hexane ringes of Toxoptera aurantii body surface also showed an attraction to all varieties. Leis axyridis var. spectabilis was the most sensitive variety.

Soil contamination with geohelminth ova in a tea plantation.

OBJECTIVE: To determine the degree of soil pollution in an area of high prevalence and the risk of exposure to geohelminth infection. DESIGN: A cross-sectional study. SETTING: A division of a large tea plantation in Kandy. SUBJECTS: All persons resident in one division of the tea plantation, above the age of 1 year. MEASUREMENTS: Prevalence of geohelminth infection measured by microscopic examination of faeces of the study group. Soil samples from 38 specific locations, obtained on 3 different occasions, about 6 months apart, analysed for helminth ova. The average temperature, rainfall and the number of rainy days during the sampling period. RESULTS: Of the 99 subjects of the study group, 77.7% were positive for at least one geohelminth infection. The prevalence rates for ascariasis, trichuriasis and hookworm infection were 54%, 28%, and 22% respectively. At the first sampling, 10.5% of the soil samples were positive for helminth ova with 7.8% and 2.6% samples showing Ascaris and Necator ova respectively. At the second sampling of the same locations nearly 40% of the samples were positive for helminth ova with 21.6%, 7.8% and 10.5% of the samples positive for Ascaris, Trichuris and Toxocara ova respectively. At the third sampling, 26.2% of the samples were positive with 21%, 2.6% and 2.6% of the samples positive for Ascaris, Trichuris and Toxocara ova respectively. Necator ova were found only once in 2.6% of the samples of the first sampling. The larger number of samples positive for helminth ova were in the second and the third samplings which coincided with high rainfall in the pre-sampling period. CONCLUSIONS: The prevalence of geohelminthiases among the study group was high. Results of soil analysis confirmed continued contamination of soil in and around the home gardens and the work areas with human and dog faeces. With the knowledge of exact locations of soil pollution it was possible to advice the management on remedial measures.