Due to its rapid development of resistance to nearly all arrays of acaricide, Tetranychus urticae is extremely hard to control using conventional acaricides. As an alternative control measure of acaricide-resistant mites, RNA interference (RNAi)-based methods have recently been suggested. A double-stranded RNA (dsRNA) delivery method using multi-unit chambers was established and employed to screen the RNAi toxicity of 42 T. urticae genes. Among them, the dsRNA treatment of coatomer I (COPI) genes, such as coatomer subunit epsilon (COPE) and beta 2 (COPB2), resulted in high mortality [median lethal time (LT50) = 89.7 and 120.3h, respectively]. The transcript level of the COPE gene was significantly (F3,9 = 16.2, P =0.001) reduced by up to 24% following dsRNA treatment, suggesting that the toxicity was likely mediated by the RNAi of the target gene. As a toxicity enhancement strategy, the recombinant dsRNA was generated by reciprocally recombining half-divided fragments of COPE and COPB2. The two recombinant dsRNAs exhibited higher toxicity than the respective single dsRNA treatments as determined using LT50 values (79.2 and 81.5h, respectively). This finding indicates that the recombination of different genes can enhance RNAi toxicity and be utilized to generate synthetic dsRNA with improved RNAi efficacy.

Toxicity test of contaminate soil is very complex because of differential bioavailability in the soil. Therefore, bioavailability of metals in soil is a major factor influencing estimates of toxicity. In this study, the two major test was conducted. First, the toxicity of arsenic for the Collembola, Paronychiurus kimi, was assessed by determining the effects of increasing arsenic concentration on survival, reproduction and body concentration of As in five forested soils with different available phosphate and oxide-metal concentration. Second, the sequential extraction procedure (SEP) for arsenic by choosing extraction reagents commonly used for sequential extraction of metals was tested. The EC50 based on total As concentration in soil was estimated respectively. The available phosphate and oxide-metal concentration in soil influenced on As fraction in soil. Especially, As in soil which is non specifically and specifically sorbed (fraction 1, 2) has strong correlation with available phosphate and oxide-metal concentration (p<0.05). The toxicity is more higher in the soil with high available phosphate and low oxide-metal concentration. In addition, the high arsenic concentration in fraction which is amorphous and poor-crystalline hydrous oxide of Fe and Al (fraction 3) had effect to the toxicity. As a result, the toxicity of As is related with As concentration in fraction 1, 2 and 3 and the soil properties and the arsenic fractionation in soil have a influence on the bioavailability and toxicity.

비티 포자와 Xenorhabdus nematophila (Xn)의 배양액을 혼합하여 비티플러스가 개발되었다. 높은 살충력에도 불구하고 비티플러스는 다 양한 해충에 대한 넓은 살충범위를 보이지 않는 한계가 있다. 본 연구에서는 Xn 대사물질 첨가를 통한 파밤나방과 같은 비감수성 해충에 대한 비 티플러스의 살충력 향상에 초점을 맞추었다. Xn의 주요 대사물질인 oxindole (OI)과 benzylideneacetone (BZA)는 비티의 살충력을 향상시킨 다고 보고되었다. 본 연구에서 OI 또는 BZA의 첨가는 비티플러스의 살충력을 향상시켰다. 그러나 동결건조된 Xn 배양액의 첨가는 보다 낮은 농 도의 OI 또는 BZA로도 충분히 비피플러스의 살충력을 상승시켰다. HPLC 분석에서 Xn 배양액에 최소 12개의 대사물질이 포함되어 있는 것을 확인하였다. 이러한 결과는 OI와 BZA 외에도 Xn 대사물질에 생리활성물질이 존재하는 것을 제시한다.

This study is the first report about the toxicity of pesticides to the mycophagous predator, I. koebelei, of powdery mildew of agricultural crops. Pesticides we tested are composed of synthetic and environmental-friendly products and being used conventionally for the control of insect or microbial pests on cucumber in Korea. our study was conducted to determine the relative toxicities of several pesticides used in Korea cucumber production to mycophagous natural enemy, I. koebelei and to provide a background for implementation of integrated powdery mildew management programs. Based on IOBC classification, three insecticides, bifenthrin + imidacloprid, acetamiprid + indoxacarb, acetamiprid + etopheprox are classified as having a Class 4 (harmful). Spiromesifen showed the low toxicity to the survival and the fecundity of I. koebelei when this chemical had been exposed to 3rd larva or newly emerged adult via feeding with cucumber powdery mildew. However, pyriproxyfen not only decreased the fecundity of female adult but also strongly prohibited from pupation. Many commercial biological or botanical pesticides can restrict the population of I. koebelei. However, Q pact (a.i. Ampelomyces quisqualis 94013), Top seed (a.i. Paenibacillus polymyxa AC-1), BT one (Bacillus thuringiensis) and Solbitchae (insecticidal microorganism) had no toxicity to I. koebelei when this chemical had been exposed to 3rd larva or newly emerged adult feeding with cucumber powdery mildew.

An assessment was made of the fumigant toxicity of 20 constituents from catnip oil and another additional five previously identified compounds of the oils and control efficacy of three experimental spray formulations containing catnip oil (1, 0.5 and 0.1% sprays) to females from B- and neonicotinoid-resistant Q-biotypes of Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae). Spathulenol (LC50, 0.39 μg/cm3) and thymol (0.45 μg/cm3) were the most toxic compounds, followed by carvacrol, α-terpineol, nerol, linalool, menthol and eugenol toward Q-biotype females (0.85–1.24 μg/cm3). The toxicity of these compounds was virtually identical toward both biotype females, indicating that the terpenoids and the insecticides (neonicotinoids and dichlorvos) do not share a common mode of action or elicit cross-resistance. The 0.5% spray of oil formulation resulted in > 80% mortality toward both biotype females. Global efforts to reduce the level of toxic synthetic insecticides in the agricultural environment justify further studies on catnip oil-derived materials as potential contact-action fumigants for the control of B. tabaci populations.

This study was aimed at assessing the fumigant toxicity to adult Thrips palmi and Orius strigicollis of basil (Ocimum basilicum) essential oil compounds and structurally related compounds using vapour-phase toxicity bioassays. Results were compared with those for dichlorvos. Against adult T. palmi, linalool (LD50, 0.0055 mg cm–3) was the most toxic fumigant and was 15.2–fold more effective than dichlorvos (0.0837 mg cm–3). Strong fumigant toxicity was also observed in pulegone (0.0095 mg cm–3), (±)-camphor (0.0097 mg cm–3) and 1,8-cineole (0.0167 mg cm–3). Moderate toxicity was produced by camphene, 3-carene, (–)-menthone, (+)-α-pinene, (+)-β-pinene, α-terpineol and (–)-α-thujone (0.0215–0.0388 mg cm–3). Against adult O. strigicollis, dichlorvos (LD50, 9.0 ×10–10 mg cm–3) was the most toxic fumigant, whereas the LD50 values of these compounds ranged from 0.0127 to >0.23 mg cm–3. Based upon selective toxicity ratio (STR, O. strigicollis LD50/T. palmi LD50), the compounds described (STR, 0.7–>10.7) are more selective than dichlorvos (STR, 10.8×10–9 mg cm–3). Basil oil compounds described merit further study as potential insecticides for control of T. palmi in greenhouses because of their generally lower toxicity to O. strigicollis and their greater activity as a fumigant than dichlorvos.

Essential oils from 22 plant species were tested for their insecticidal activities against spotted wing drosophila (SWD), Drosophila suzukii Matsumura, using fumigation and contact bioassay. Responses varied with the essential oils, their constituents and concentrations. Strong fumigation activities were observed from the essential oils of wintergreen (Gaultheria fragrantissima), croton (Croton anisatum) and star anise (Illicium verum). Strong contact activities were observed in the essential oils of oriental sweetgum (Liquidambar orientalis), cassia (Cinnamomum cassia), damask rose (Rosa damascena) and sandalwood (Santalum album). The compositions of these selected essential oils were identified using gas chromatographymass spectrophotometer. The compounds identified were tested individually for their insecticidal activities against SWD. Responses varied by doses for each compound. The results showed that natural materials are as effective as synthetic pesticides.

The cold shock of spermatozoa is associated with oxidative stress induced by reactive oxygen species. This study was conducted to evaluate the toxicity of natural antioxidant green tea extract (GTE) in lactose-egg yolk (LEY) extender during boar sperm cooling prior to freezing. Spermatozoa were cooled to 5℃ for 3 h in LEY extender containing 0 (control), 1, 10, 100 or 1,000 mg/l of GTE, re-suspended with LEY-glycerol-Equex extender and cooled at 5℃ for 30 min. Sperm progressive motility, viability and phosphatidylserine (PS) translocation were evaluated. PS translocation was assayed by flow cytometry using Annexin V-FITC apoptosis detection kit. The sperm function including progressive motility, viability and PS translocation was not significantly different regardless of GTE concentrations (P>0.05). In conclusion, this study demonstrated non-toxicity of GTE supplement in LEY extender during sperm cooling.

The safety of a new natural plant composition (ADP) was assessed on the genotoxicity study and 14-day repeat dose toxicity study. ADP contains a mixed water extract obtained from the mixture of Phellodendron cortex (Phellodendron amurense) and Anemarrhena rhizoma (Anemarrhena asphodeloides), and poses the contractile properties mediated by alpha-adrenoceptor of the prostate and urethra as well as antioxidant and anti-inflammatory properties. In order to evaluate genetic safety, in vivo micronucleus test was performed in ICR mice orally administered with three dose levels of 1250, 2500, 5000 mg/kg body weight, and vehicle and positive control. In the 14 days study, Sprague-Dawley rats were treated with ADP at the dose levels of 500, 1000, 2000 mg/kg once a day, and clinical signs, body weights, hematology, serum biochemistry, necropsy findings and organ weights were monitored and examined. In experimental results, ADP treatment, compared with vehicle control, did not induce the micronucleated erythrocytes from mouse bone marrow. In the 14 days study, any significant and toxicological differences in all measurements of parameters were not observed in ADP treatment groups of animals, compared with vehicle treatment. The No-Observed-Adverse-Effect-Level (NOAEL) of ADP in the 14 days study was determined to be greater than 2000 mg/kg/day in both sexes.

몇 가지 살충제에 대한 독성을 썩덩나무노린재를 대상으로 단감원에서 잔효독성으로 검증하였고 끈끈이 트랩에 유인된 뚱보기생파리 성충 을 대상으로 직접분무처리 방식으로 검증하였다. 썩덩나무노린재는 흑색유아등으로 채집하였고 뚱보기생파리 성충은 갈색날개노린재 집합페로 몬인 methyl-(E,E,Z)-2,4,6-decatrienoate을 사용해서 끈끈이트랩으로 포획하였다. 비펜트린수화제, 뷰프로페진･디노테퓨란수화제, 클로티아 니딘액상수화제, 디노테퓨란수화제, 티아메톡삼입상수화제 등 5종의 살충제 중에서 비펜트린수화제가 72시간 케이지 내 잔효독성 검정결과 썩 덩나무노린재에 대해 93.1%의 사충율을 나타내었고 여타 살충제들은 다양한 사충율을 보였다. 뚱보기생파리에 대한 사충율은 55.3~74.3%이 었다.

The insecticidal activity of cinnamon essential oils, cinnamon bark, cinnamon technical, cinnamon green leaf oils and their constituents and structurally related compounds against citrus flatid planthopper, Metcalfa pruinosa, was examined using a direct contact application. At 500 mg litre-1 very strong mortality (100%) was observed in cinnamon technical, cinnamon bark, cinnamon green leaf, mortality against flatid planthopper M. pruinosa. The cinnamon oils constituents were identified by GC-MS. The active principles were determined to be hydrocinnamic acid (24 h LC50, 30.66 mg/L), geranic acid (24 h LC50, 31.23 mg/L), cinnamaldehyde (24 h LC50, 32.65 mg/L), hydrocinnamaldehyde (24 h LC50, 39.11 mg/L) and trans-cinnamaldehyede (24 h LC50, 39.54 mg/L) were the most toxic against both nymph and adult of citrus flatid planthopper, M. pruinosa. The moderate activity was observed with cinnamyl acetate, dibutyl pthalate, anethole, α -cyano cinnamic acid, cinnamyl alcohol, methyl cinnamaldehyde, eugenol, 2.4-dihydrocyl cinnamic acid, bornyl acetate (24 h LC50, 53.35- 97.17 mg/L) respectively. The other constituents were showed less or no activity against adult of M. pruinosa. Global efforts to reduce the level of highly toxic synthetic insecticides in the agricultural environment justify further studies on the active cinnamon oils active principles act as potential insecticides for the control of M. pruinosa populations as direct spray with contact action.

Double-stranded RNA(dsRNA) had been used to specitically suppress target gene expression at post-tanscription level. Injection of dsRNA to hemocoel is the most efficient to knockdown target mRNA. However, some insects have shown to be susceptible to feeding dsRNA. Spodoptera exigua was susceptible to dsRNA at oral treatment. Especially dsRNA specific to β-integrin was potent to survival of S.exigua larvae. This study advanced our dsRNA application technology by generating recombinant E.coli expressing dsRNA specific the β-integrin. A recombinant vector L4440 was constructed with a partial β-integrin gene under T7 RNA polymerase promoter. The recombinant vector was used to transform HT115 competent cells of E.coli. The transformed E.coli expressed the dsRNA. The production of dsRNA was proportional to the bacterial number. By feeding the recombinant E.coli, S.exigua underwent significant mortality. By adding E.coli expressing Cry1Ca Bt toxin to E.coli expressing dsRNA, S.exigua exhibited highly enhanced mortality. This study suggests a possibility to use a recombinant E.coli expressing dsRNA to control S.exigua.

Behavioral reaction can be measured as a sensitive endpoint for sublethal toxicity of copper, and can be used to obtain easily and quickly. Also behavioral endpoints may serve as a more insightful evaluation tool of the ecological effects of toxic chemicals. In this study, four invertebrates in freshwater which are usually used indicate species for water quality were selected as test species (Chironomus riparius, Heterocypris incongruens, Daphnia magna, and, Triops longicaudatus). Each test species was exposed to copper for 6 hours, and total distance, velocity, and, turn angle were measured for 1 hour using video analysis system (Ethovision : Noldus Information Technology) in laboratory condition. Each endpoints reflected effect of copper toxicity appropriately for all test species. These endpoint have possibility that can be used to identify characteristic behavioral responses to a metal toxicity. We viewed this study as a preliminary experiment for future research to investigate the significance of behavioral endpoints to various toxic chemicals.

Six of cadherins have been selected from the P. xylostella genome 52 open reading frames are annotated as cadherin-like genes. Compared to other 5 cadherins of P. xylostella (PxCads), PxCad1 has the highest homology with other lepidopteran insect cadherins and PxCad1 was expressed in all developmental stages specially in gut tissue. Expression of PxCad1 was suppressed by feeding its specific double-stranded RNA (dsRNA, 150ng/larva) and treatment of dsPxCad1 significantly reduced susceptibility to Bt Cry1Ac toxin. To confirm the specific interaction between PxCad1 and Cry1Ac, a toxin-binding assay was performed using enzyme-linked immunosorbent assay (ELISA). The ELISA indicates that BBMV extracted from PxCad1-silenced P. xylostella have significantly lower binding activity to active form of Cry1Ac than control BBMV. Moreover, the analysis of the binding parameters showed that the toxin affinity (Kd) of the control BBMV extract (BBMV-dsCON) was 6.08 ± 0.84 nM, which was not much different to the affinity value (6.72 ± 0.81 nM) of the dsPxCad1 treatment. However, there was a remarkable difference in number of binding sites (Bmax), in which BBMV-dsCON extract had 1.61 ± 0.04, but the BBMV-dsPxCad1 extract had 0.88 ± 0.02. Taken together, these results are suggest that PxCad1 is a functional receptor for Cry1Ac toxicity against P. xylostella larva.

Oral exposure of humans by excess amounts of arsenic may cause disturbances of the reproductive system. In the present study, such exposure was modelled in rats, with the support of sperm principal parameters and histopa-thological observations. Male Sprague–Dawley rats were randomly divided into three groups where the group I was served as a normal control, group II was received sodium meta-arsenite as arsenic (10 mg/kg b.w/day) and a combi-nation of sodium meta- arsenite and sodium selenite (3 mg/kg b.w/day) in group III. After 6 weeks, there was no significant change in testis weight and in total motility of all the three experimental groups, whereas, rapid moving spermatozoa, moderately moving spermatozoa and slow moving spermatozoa were significantly decreased in arsenic treated rats as compared to control rats. The other sperm principal parameters like progressiveness, average path velocity, straightness linear velocity (VSL), curvilinear velocity (VCL), straightness, linearity sperm head elongation ratio, area, linearity amplitude of lateral head department (ALH) and beat cross frequency (BCF) were found to be reduced in arsenic intoxicated rats. These results are not correlated with the histological studies. On oral admini-stration of selenium ameliorated the adverse effects of arsenic as compared to arsenic alone treated rats. Our findings clearly demonstrate that administration of selenium could prevent some of the deleterious effects of arsenic in the testis.

Bemisia tabaci is a serious pest in various horticultural crops in the world. Due to use of chemical pesticide for their management they develop pesticide resistance and environmental contamination. It is necessary to develop alternative bio-pesticides using natural products from plants and natural enemies. Nicotiana benthamiana is a variety of wild tobacco plants and produce acyl sugars from glandular trichomes in the leaves. When adult whiteflies were reared with fresh N. benthamiana leaves, they were completely dead within 84 h. Oral feeding of 20% N. benthamiana extracts using ethanol and water showed complete mortality of whiteflies within 48 hours. Spray of N. benthamiana extracts into the leaves was lethal to eggs but not to nymphs of whiteflies. Further, tomato plants sprayed with N. benthamiana extracts were highly repellent to adult whiteflies. Quantitative real-time PCR indicated that the expression of various genes of B. tabaci was changed by oral feeding of N. benthamiana extract. This study suggests N. benthamiana extract is a useful for the control of whiteflies and can be used as an alternative natural pesticide for the whitefly management.

본 연구는 Spragye-Dawely 계통의 암컷 랫드에서 종합 비타민의 반복경구투여 독성평가와 대식세포 Raw 264.7 세포의 NO 및 TNF-α assay를 통한 면역 활성을 평가하기 위해서 실시하였다. 종합비타민을 대식세포의 활성능을 측정하기 위해 Raw 264.7 세포에서 NO와 TNF-α의 생성을 측정하였다. 종합비타민을 대식세포에 24시간 처리한 결과 대조군과 비교 시 NO와 TNF-α가 유의적으로 상승하였다. 이 결과 종합비타민이 대식세포인 Raw 264.7 세포를 활성화시키는 것으로 사료된다. 또한 랫드에서 종합비타민의 독성평가를 위하여 랫드에 종합비타민을 0.24 g/ kg, 1 g/kg 그리고 2 g/kg을 4주 동안 경구투여를 하였다. 종합비타민의 안전성을 확인하기 위해 다음과 같은 관찰 및 검사를 하였다. 검사항목으로는 체중과 사료 섭취량, 임상증상, 혈청생화학적 검사를 관찰한 결과 대조군과 투여군을 비교 시 유의적인 변화가 나타나지 않았다. 따라서 종합비타민은 생리대사에 무해하며 면역증강의 효과를 나타내는 것으로 사료된다.

The insecticidal activity of 120 plant essential oils and control efficacy of six experimental spray formulations (SF) containing the oils (SF-0.25, 0.5, 1, 2.5, 5 and 10% sprays) against both nymph and adult of citrus flatid planthopper, Metcalfa pruinosa, was examined using a direct contact application. Reponses varied according to dose (1000 mg litre-1 and 500 mg litre-1). Based on 24 h exposure in leaf dipping assay at 1000 mg litre-1 strong mortality was observed in more than 19 essential oils among 130 was screened. At 500 mg litre-1 very strong mortality (100%) was observed in cinnamon technical, cinnamon green leaf, cinnamon #500, cassia tree, citronella java and penny royal followed by origanum, thyme white, grape fruit, savory, fennel sweet, aniseed and cinnamon bark (93.3- 80%) showed considerable moratality against nymphs of M. pruinosa. The moderate mortality was found in thyme red, tagette, calamus, lemoneucalptus and geranium (73.3-60%). The other oil has low or very low mortality against M. pruinosa. The oil applied as SF-10% sprays provided 100% mortality against adult M. pruinosa. In particular cinnamon technical showed very strong (>SF-0.5= 100%) effect followed by cinnamon #500 (>SF-2.5=100%), cinnamon green leaf (>SF-2.5=100%) and penny royal (>SF-2.5=100%) respectively. Global efforts to reduce the level of highly toxic synthetic insecticides in the agricultural environment justify further studies on the active essential oils as potential larvicides for the control of M. pruinosa populations as direct spray with contact action.

An assessment was made of the toxicity of basil, Ocimum basilicum, essential oil, 11 basil oil constituents, seven structurally related compounds and another 22 previously known basil oil constituents and the control efficacy of four experimental spray formulations containing the oil (1, 2, 3 and 4% sprays) to adult American house dust mites, Dermatophagoides farinae. Results were compared with those of two conventional acaricides benzyl benzoate and N,N-diethyl-3- methylbenzamide (deet). The active principles were determined to be citral, α -terpineol and linalool. Citral (24 h LC50, 1.13 μg/cm2) and menthol (1.69 μg/cm2) were the most toxic compounds, followed by methyl eugenol (5.78 μg/cm2). The toxicity of these compounds was more toxic than that of benzylbenzoate(LC50, 8.41 μg/cm2) and deet (37.67 μg/cm2). These compounds were consistently more toxic in closed versus open containers, indicating that their mode of delivery was largely a result of vapour action. Basil applied as 3 and 4% experimental sprays provided 97 and 100% mortality against the mites respectively, whereas permethrin (cis:trans, 25:75) 2.5 g/l spray treatment resulted in 17% mortality. Reasonable mite control in indoor environments can be achieved by spray formulations containing the basil oil (3 and 4% sprays) as potential contact-action fumigants.

The present study investigated the urinary tract toxicity of melamine alone or in combination with cyanuric acid in rats. Male rats were orally administered melamine alone (800 mg/kg) or melamine plus cyanuric acid (50 mg/kg each) for 3 consecutive days. Although melamine treatment did not show any significant difference in body weight, kid- ney or urinary bladder weight, serum blood urea nitrogen (BUN) or creatinine levels, it caused a decrease in urinary pH and mild histopathological alterations in the kidney and urinary bladder. In contrast, co-administration of melamine and cyanuric acid induced a decrease in body weight, an increase in kidneys and urinary bladder weights, and an elevation in serum BUN and creatinine levels, which were not observed in animals treated with melamine alone. His- topathological examination showed that the incidence and severity of histopathological lesions in the kidney and uri- nary bladder were much higher than those in the melamine group. Urinalysis revealed an increase in urine occult blood, leukocytes and protein and a decrease in urinary pH. These results indicate that concomitant administration of cyanuric acid synergistically potentiated the urinary tract toxicity induced by melamine in rats. In this study, we first demon- strated the synergistic toxic effects of melamine in combina- tion with cyanuric acid on urinary bladder and urinalysis in rats.