Impact of Spodoptera littoralis (Boisd.) and Agrotis ipsilon (Hufn.) larval frass on oviposition of conspecific insects

Ethanolic extracts of larval frass of the cotton leaf worm and the black cutworm were prepared and tested to deter the eggs lay of the adults of the same insects. Two different types of food were used for larval feeding. Extracts fractions were identified using gas chromatography–mass spectrometry analysis. High concentrations were more deterrent to oviposition than low. Extracted frass of fed larvae on semi-artificial diet was more effective than others fed on natural host. Sensitivity of the black cutworm adult females to the cotton leaf worm frass extract was clearly observed at high tested concentration of L_1–3 and L₄ frass extracts which resulted completely oviposition deterrent. Several fatty acids were identified qualitatively and quantitatively in frass extracts of different larval instars of both target insects. Type and quantity of fatty acid depends mainly on larval food source and larval instar, except palmitic acid which recorded at all larval instars and food sources. Oleic acid and ethyl 9-hexadecaenoic acid were found when semi-artificial diet used as a food source while myrisitic acid was observed only in extracted frass of fed larvae on castor oil leaves.     

Increase in acid phosphatase activity in the fat body during larval and pharate pupal development in Calliphora erythrocephala

Biochemical evidence was obtained for an increase in acid phosphatase activity in the larval fat body of Calliphora erythrocephala during larval and pharate pupal instars. This observation is in conflict with published data indicating a decreasing enzyme activity in late third stage larvae. Centrifugation and filtration studies showed that the pH of the homogenisation medium has a strong influence on the solubilisation of acid phosphatase and its distribution in homogenate components. Differences in biochemical techniques including the pH value may explain the discrepancy between the published results and the present findings.The observed increase in acid phosphatase activity is related to the activity of the lysosomal system in the period immediately preceding pupal-adult apolysis.     

甜菜夜蛾不同龄期幼虫对药剂的敏感性及其与酶活力的关系

【目的】明确甜菜夜蛾Spodoptera exigua(Hübner)不同龄期幼虫对甲氨基阿维菌素苯甲酸盐和毒死蜱的敏感性及其与羧酸酯酶、多功能氧化酶、谷胱甘肽S-转移酶和乙酰胆碱酯酶活性的相关性。【方法】采用室内生物测定方法检测甜菜夜蛾不同龄期幼虫对药剂的敏感性,并检测了不同龄期幼虫体内羧酸酯酶、多功能氧化酶、谷胱甘肽S-转移酶和乙酰胆碱酯酶的活力。【结果】在所测定的5个龄期中,随龄期增加,敏感性逐渐降低,其中1~5龄幼虫对甲氨基阿维菌素苯甲酸盐的LC50分别为0.1010、0.3561、0.7568、1.4325和8.4390 mg/L,对毒死蜱的LC50分别为27.4632、46.8495、87.5222、129.3217和1 356.6770 mg/L。单头幼虫的羧酸酯酶、多功能氧化酶、谷胱甘肽S-转移酶和乙酰胆碱酯酶活力随虫龄的增加而提高,与龄期间对药剂的敏感性呈负相关(由于外源化合物摄入量减少,5龄单头幼虫的多功能氧化酶活力略有降低)。【结论】甜菜夜蛾不同龄期幼虫对药剂的敏感性存在非常明显的差异,在田间防治中,应选择对杀虫剂敏感性较高的低龄幼虫作为最佳防治时期。     

Orientation Behavior,Development and Survival of <Emphasis Type="Italic">Trichoplusia ni</Emphasis> (Lepidoptera: Noctuidae) Larvae on Cotton Expressing Cry1Ac and Cry2Ab and Conventional Cotton

This study was conducted to determine the effects of Bt cotton leaves (Bollgard II), non-Bt cotton leaves, and a mixture of Bt+non-Bt cotton leaves on larval orientation behavior, survival and development of Trichoplusia ni in the laboratory. Results indicate that in a no-choice test, more first and fifth instars remained on Bt leaves than the third instars. All larvae that remained on the leaves gradually moved to leaf edge. In the choice between a Bt and a non-Bt leaf, more first instars moved to non-Bt leaves, whereas the third and fifth instars did not show significant difference in the first 8 h, but eventually more moved to non-Bt leaves. More first instars fed non-Bt leaves than third instars and fifth instars. When larvae fed Bt leaves, 100% of first instars, 92.7% of third instars and 51.1% of fifth instars died in 108 h. Once larvae pupated, >90% developed to adults. First and third instars that fed Bt leaves developed slower but their pupae developed faster than those on Bt+non-Bt leaves, whereas fifth instars developed similar on the three types of leaves. First and third instars that fed Bt+non-Bt leaves resulted in less heavy pupae than those fed non-Bt leaves; whereas the fifth instars that survived on Bt leaves produced lighter pupae.     

利用甲基对硫磷降解菌DLL-E4消除农产品表面农药污染的研究

利用甲基对硫磷降解菌DLL-E4(Pseudomonas sp.)能高效分解甲基对硫磷的特性,研究了消除农产品表面农药残留污染的途径,采用菌体发酵液、发酵液上清液、菌体蛋白、粗酶液、添加菌体蛋白的洗涤剂以及洗涤剂,处理了小青菜、茶叶、黄瓜表面的农药残留污染。结果表明,使用发酵液和不同的酶制剂可以有效去除这些产品表面的农药残留污染,最高去除率可达100％;DLL-E4上清液中有一些酶促反应促进因子,有助于酶更好地发挥作用,洗涤剂和酶直接混合作用,有助于消除茶叶表面的农药残留,但对于黄瓜和小青菜则相反。在所有选用的模式中,使用粗酶液和菌体蛋白效果最好,对黄瓜、小青菜和茶叶合适的酶制剂浓度分别为2％、5％和10％,粗酶液同时还能降解甲胺磷、辛硫磷、毒死蜱等农药。     

Castor and camphor essential oils alter hemocyte populations and induce biochemical changes in larvae of Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae)

Two plant essential oils; camphor and castor were tested for insecticidal and antifeedant activity against the 4th instar larvae of Spodoptera littoralis, a serious pest on cotton in Egypt. Also the impact of LC₁₀ of both oils on some physiological parameters in larvae was studied by using leaf dipping technique. Analysis of both oils using GC–MS revealed several insecticidal and antifeedant compounds. Our results showed higher insecticidal activity and antifeedant index of camphor oil against S. littoralis. The LC₅₀ and the antifeedant indices were 163.1, 246.8?mg/ml and 12.69, 6.62% for camphor and castor bean oil, respectively. The total hemocyte count (THC) and differential hemocyte count (DHC) were reduced significantly after 48?h of treatment compared to controls. Both oils reduced all types of hemocytes except plasmatocytes which were reduced only by castor oil. Camphor oil decreased total proteins and carbohydrates while castor oil targeted only carbohydrate content. Both oils didn't affect the amount of total lipids. Lipase, α-amylase and glucose-6-phosphate dehydrogenase (G6PD) enzyme activities were increased significantly in larvae treated with camphor oil than other treatments. These results clearly indicate that castor and camphor oils can affect the nutritional status in S. littoralis larvae, thereby changing the internal metabolic processes in the larvae which make them as potential control agents in IPM programs against S. littoralis.     

Polyphenolic extracts from pomegranate and watermelon wastes as substrate to fabricate sustainable silver nanoparticles with larvicidal effect against Spodoptera littoralis

The agricultural wastes adversely affect the environment; however, they are rich in polyphenols; therefore, this study aimed to employ polyphenol-enriched waste extracts for silver nanoparticles synthesis, and study the larvicidal activity of silver nanoparticles fabricated by pomegranate and watermelon peels extracts (PPAgNPs and WPAgNPs) against all larval instars of Spodoptera littoralis. The polyphenol profile of pomegranate and watermelon peel extracts (PP and WP) and silver nanoparticles was detected by HPLC. The antioxidant activity was estimated by DPPH, and FARP assays and the antimicrobial activity was evaluated by disc assay. The Larvicidal activity of AgNPs against Egyptian leaf worm was performed by dipping technique. The obtained AgNPs were spherical with size ranged 15–85 nm and capped with proteins and polyphenols. The phenolic compounds in silver nanoparticles increased about extracts; therefore, they have the best performance in antioxidant/reducing activity, and inhibit the growth of tested bacteria and yeast. The PPAgNPs were the most effective against the first instar larvae instar (LC₅₀ = 68.32 µg/ml), followed by pomegranate extract with (LC₅₀ = 2852 µg/ml). The results indicated that obvious increase in polyphenols content in silver nanoparticles enhance their larvicidal effect and increasing mortality of 1^st larval of S. littoralis Egyptian leafworms causing additive effect and synergism. We recommend recycling phenolic enriched agricultural wastes in producing green silver nanoprticles to control cotton leafworm that causes economic loses to crops.     

花椒叶浸提液对土壤微生物数量和土壤酶活性的影响

通过用花椒叶浸提液浇灌盆栽花椒幼苗,研究浸提液对土壤酶和土壤微生物的影响.结果表明, 花椒叶浸提液使根际土中细菌、真菌和放线菌数量以及微生物总数均有不同程度的减少,根际土中真菌和放线菌的数量变化呈降-升-降-升的趋势.20、60和80 g·L^-1浓度的叶浸提液使非根际土中细菌的数量显著增加21.59%、107.55%和8.62%,而40 g·L-1浓度的叶浸提液则使非根际土中细菌数量显著降低22.56%.叶浸提液使根际土蛋白酶、蔗糖酶和酸性磷酸酶活性明显低于非根际土相应的酶活性,而过氧化氢酶和多酚氧化酶活性则显著升高.土壤的蛋白酶活性与蔗糖酶活性呈显著正相关,与土壤放线菌数量呈显著负相关;多酚氧化酶活性与蔗糖酶活性呈显著负相关,与细菌、真菌、放线菌以及微生物总数呈显著正相关;放线菌只与蛋白酶、多酚氧化酶、蔗糖酶3种酶活性及真菌呈显著相关,与过氧化氢酶、酸性磷酸酶以及细菌和微生物总数的相关性均不显著.     

甜菜夜蛾对不同寄主植物的产卵和取食选择

为了探讨甜菜夜蛾Spodoptera exigua对不同寄主植物的产卵选择及成虫产卵选择与幼虫取食选择间的关联度, 本研究选取玉米、豇豆、甘蓝、黄瓜、棉花、辣椒和番茄7种植物进行了选择性和非选择性实验研究, 并采用Y型嗅觉仪测定了成虫对其中3种寄主植物及其挥发物抽提物的趋性。结果表明：在田间非选择性实验中, 甜菜夜蛾在不同寄主植物上的落卵量依次为：玉米>辣椒>棉花>黄瓜、豇豆、番茄>甘蓝。Y型嗅觉仪的行为测定表明, 雌成虫对玉米及其挥发物抽提物的趋性最强, 黄瓜次之, 对甘蓝的趋性最弱, 这与雌虫的产卵选择性一致。不同龄期甜菜夜蛾幼虫对寄主植物的取食选择性有所不同, 且随观测时间的延长有所改变;低龄幼虫对豇豆、玉米和黄瓜的选择性较强, 对甘蓝、番茄、辣椒和棉花的取食选择性则较弱, 高龄幼虫对辣椒也具有较强的选择性;5龄幼虫对寄主植物的选择性不如低龄幼虫明显。结果显示, 甜菜夜蛾对不同寄主植物的产卵选择性显著不同, 植物抽提物在雌成虫的产卵选择中具有重要作用, 甜菜夜蛾对寄主植物的产卵选择性和幼虫取食选择性并不一致。     

Changes of acid phosphatase content and activity in the fat body and the hemolymph of the flesh fly Neobellieria (sarcophaga) bullata during metamorphosis

Changes in the specific and total activity of the lysosomal marker enzyme acid phosphatase (Acph) and in the amount of enzyme protein were examined in the fat body and the hemolymph from the last larval molt to the larval-pupal apolysis. The specific activity showed minor changes during the last larval period. In contrast, the total activity of the enzyme was low during the feeding period and higher during the wandering stage and strikingly increased at the time of puparium formation. We purified a protein having para-nitrophenyl phosphate phosphatase (Acph) activity and raised antisera against it. The amount of Acph protein in the fat body and hemolymph was examined using an ELISA. The specific Acph content showed little variation, but the total amount of the enzyme protein showed a stepwise increase in both organs during last larval stage and was markedly elevated in the pupal stage in the fat body. In contrast, a considerable decrease in the amount of Acph protein was observed in the hemolymph during this period. These data were in agreement with immunohistochemical observations showing an accumulation of the enzyme protein in fat body cells during the prepupal stage with a concomitant disappearance of the enzyme from the hemolymph. Inhibition of ecdysteroid secretion by water stress prevented the changes both in total enzyme activity and in the amount of Acph protein. However, Acph protein content and enzyme activity could be restored when the water stress was followed by a 20-hydroxyecdysone (20-HE) treatment. Taken together, our data show that Acph is secreted by fat body cells into the hemolymph during the larval stage, where it is stored in an inactive form. Increase in the 20-HE titer at the end of last larval stage reverses this process, and the enzyme is taken up by the fat body cells, where it becomes activated and appears in auto- and heterophagic vacuoles. Arch. Insect Biochem. Physiol. 34:369–390, 1997. © 1997 Wiley-Liss, Inc.     

Foraging and growth responses of cotton armyworm Spodoptera litura to the biophysical characteristics of five cotton varieties

This study investigated the morphological and biochemical characteristics of the CB1, CB3, CB5, CB8 and CB12 cotton varieties and evaluated their effects on third instar larval movement, and body weight of the cotton armyworm Spodoptera litura at different developmental stages. The cotton varieties differed in their plant architecture, branching, stem color and hairiness, leaf color and hairiness, leaf trichome density, flower color, numbers of leaves and bolls per plant, concentrations of protein and starch, and boll length, width and weight. The CB1 and CB3 varieties possessed significantly higher trichome densities, while CB8 produced larger and heavier bolls. Boll bearing was found to be highest in CB1 and lowest in CB8. Biochemical analyses indicated the highest percentage of protein in CB5, and of starch in CB8; concentrations of both were lowest in CB12. Cotton varieties did not affect larval foraging, but their abundance on leaflet, mature and square differed significantly. Analysis of the growth response parameters of S. litura as a result of feeding on the tested varieties revealed that larval instars, pupae and adult moths attained the highest body weight on CB8, followed by CB5, and the lowest weight on CB12. Collectively, the results of this study show that the CB5 and CB8 varieties are favorable host‐plants for cotton armyworms; therefore, these varieties are the least suitable for cultivation.     

Induction of systemic resistance to bacterial blight caused by Xanthomonas campestris pv. malvacearum in cotton by leaf extract from a medicinal plant zimmu (Allium sativum L. × Allium cepa L.)

Abstract

Aqueous extract (10%) from leaves of zimmu (Allium sativum L. × Allium cepa L.) when applied as foliar spray to first and second leaves of cotton plants induced systemic resistance in third and fourth leaves to a challenge infection with Xanthomonas campestris pv. malvacearum and reduced the number of lesions by up to 73% compared with water-treated control plants. The treated leaves exhibited significantly high activity of enzymes phenylalanine ammonia-lyase, peroxidase and polyphenol oxidase along with rapid accumulation of phenolics. The activities of chitinase and β-1,3-glucanase were greatly elevated in treated plants as compared to water-treated controls. An 11-fold increase in chitinase activity was evident 4 d after treatment. Western blot analysis revealed that a chitinase with an apparent molecular weight of 58 kDa that cross-reacted with a barley chitinase antiserum was induced in cotton leaves 3 d after treatment and the maximum induction of this chitinase was detected 4 d after treatment. The present study provides evidence for the induction of biochemical defence mechanisms in cotton leaves after treatment with leaf extract from zimmu.     

二斑叶螨危害对棉花应激防御酶活性的影响

探明二斑叶螨取食对棉花叶片内应激防御性酶活性的影响。在棉苗5-6叶期接种不同数量(10-40头)的二斑叶螨雌成虫,取食1-4 d后采摘不同处理的叶片,利用分光光度计测定过氧化氢酶(CAT)、过氧化物酶(POD)、多酚氧化酶(PPO)、脂氧合酶(LOX)和苯丙氨酸解氨酶(PAL)活性。结果表明二斑叶螨取食后,棉花叶片CAT活性显著升高,且随诱导时间的延长其活性变化呈先上升后下降的趋势,为害3 d时,10、20、H(每株接棉蚜跟叶螨各40头)头/叶的CAT活性达到最高峰,显著高于对照(P0.05);POD、PAL活性也随诱导时间的延长和螨口数量的增加呈上升趋势,为害4 d时达到最高峰;而PPO、LOX活性随着时间的增加呈现上升后下降趋势,且为害2 d时达到最高峰,仍显著高于对照(P0.05)。棉叶应激防御酶活性随着二斑叶螨为害时间与螨口密度的增加而增加,表明二斑叶螨的危害胁迫能诱导寄主植物产生一系列应激生化反应,不同的酶活性表现有所不同。     

Differential hormone regulation of acid DNase activity in the testes and fat body of Spodoptera litura (Lepidoptera-Insecta).

Acid DNase activity in the testes and fat body is high during the early larval instars which may be correlated with the extensive cell division seen in both the tissues during these stages. The increased enzyme activity, observed in the testes of the pupal stage, might be involved in the in vivo degradation of DNA in a large number of degenerating spermatocysts which occur during this stage. Total activity of acid DNase in the fat body is highest in pupal stage. Like acid phosphatase, this enzyme may also be involved in the process of remodelling of the fat body during metamorphosis. 20-Hydroxyecdysone (20-HE) does not have any effect on acid DNase activity in the testes but it alters the enzyme activity in the fat body. Juvenile hormone-I (JH-I) has no effect on the enzyme activity in the fat body.     

CO_2浓度升高对粘虫幼虫体内酶活性的影响

【目的】阐明大气CO_2浓度升高对粘虫Mythimna separata(Walker)幼虫不同龄期体内酶活性的影响机制。【方法】测定分析了人工气候箱内不同CO_2浓度(800mL/L和400mL/L)条件下,粘虫幼虫体内4种解毒酶[羧酸酯酶(Car E)、乙酰胆碱酯酶(Ach E)、微粒体多功能氧化酶(MFO)和谷胱甘肽-S-转移酶(GST)]、3种保护酶[超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)]和4种蛋白酶(总蛋白酶、强碱性类胰蛋白酶、弱碱性类胰蛋白酶和类胰凝乳蛋白酶)活性的变化。【结果】(1)粘虫不同龄期幼虫体内4种解毒酶的活力均随着CO_2浓度升高而显著上升(P<0.05);(2)粘虫幼虫体内的保护酶中SOD酶活性在CO_2浓度为800mL/L条件下低于对照(CO_2浓度为400mL/L)条件下的酶活性,其中4至6龄幼虫的下降趋势较为显著(P<0.05),而POD、CAT酶活性均呈上升趋势,800mL/L浓度下的CAT活力均显著高于对照(P<0.05);(3)高CO_2浓度条件下粘虫总蛋白酶的酶活性随着龄期的增加呈现先上升后下降的趋势,强碱/弱碱性类胰蛋白和类胰凝乳蛋白3种酶的比活力均低于400mL/L CO_2浓度下的比活力,其中类胰凝乳蛋白酶比活力显著低于对照条件下的比活力(P<0.05)。【结论】在高CO_2浓度条件下,粘虫幼虫体内的大多数酶系活力呈现下降趋势,大部分解毒酶和蛋白酶活性受到抑制,SOD、CAT和POD 3种保护酶的动态平衡受到干扰而影响其正常的生理代谢,机体受损伤并随龄期的增加而加重。     

Mass production of aphicidal Beauveria bassiana SFB-205 supernatant with the parameter of chitinase

Beauveria bassiana SFB-205 supernatant can effectively control cotton aphid populations, which is closely associated with its chitinase activity. The present work extends to optimizing a culture medium to produce more efficacious supernatant in flask conditions, followed by scale-up in 7 L, 300 L and 1.2 KL fermentors with the parameter of chitinase. In flask conditions, a combination of soluble starch and yeast extract produced the greatest amount of chitinase (5.1 units/ml) and its supernatant had the highest aphicidal activity. An optimal quantitative combination of the two substrates, estimated by a response surface method, enabled the supernatant to have 15.7 units/ml of chitinase activity and 3.7 ml/l of median lethal concentration (LC50) of toxicity against cotton aphid adults in laboratory conditions. In the scale-up conditions, overall supernatant had 25-28 units/ml of chitinase activity. Decrease in pH and limitation of dissolved oxygen (DO) during cultures were significantly related to the yield of chitinase. These results suggest that the substrate-dependent chitinase production can be background information for optimizing a culture medium, and pH and DO are critical factors in maximizing the production in scale-up conditions.     

Spatial and temporal dynamics of hotspots of enzyme activity in soil as affected by living and dead roots—a soil zymography analysis

Aims

Hotspots of enzyme activity in soil strongly depend on carbon inputs such as rhizodeposits and root detritus. In this study, we compare the effect of living and dead Lupinus polyphyllus L. roots on the small-scale distribution of cellulase, chitinase and phosphatase activity in soil.

Methods

Soil zymography, a novel in situ method, was used to analyze extracellular cellulase, chitinase and phosphatase activity in the presence of i. living L. polyphyllus roots prior to shoot cutting and ii. dead/dying roots 10, 20 and 30 days after shoot cutting.

Results

After shoot cutting, cellulase and chitinase activities increased and were highest at the root tips. The areas of high cellulase and phosphatase activity extend up to 55 mm away from the root. Moreover, we observed microhotspots of cellulose, chitinase, and phosphatase activity up to 60 mm away from the next living root. The number and activity of microhotspots of chitinase activity was maximal 10 days after shoot cutting.

Conclusions

The study showed that young root detritus stimulates enzyme activities stronger than living roots. Soil zymography allowed identification of microhotspots of enzyme activity up to several cm away from living and dying roots, which most likely were caused by arbuscular mycorrhizal fungi.     

Feeding behaviour of Helicoverpa armigera larvae on insect-resistant transgenic cotton and non-transgenic cotton

Abstract: Feeding behaviour of Helicoverpa armigera Hübner (Lep.; Noctuidae) larvae on non‐transgenic Bacillus thuringiensis (Bt) cotton (Gossypium hirsutum L.), Zhong 30, and transgenic cowpea trypsin inhibitor (CpTI)‐Bt cotton, SGK 321, and non‐transgenic cotton, Shiyuan 321, was investigated in both choice tests and no‐choice tests. The results of choice tests suggested that neonates have the ability to detect and avoid transgenic cotton. In the choice tests of neonates with both transgenic and non‐transgenic cotton leaves, a significantly greater proportion of larvae and higher consumption were observed on non‐transgenic cotton than on the transgenic Bt or CpTI‐Bt cotton. In the choice tests with leaves of two transgenic cotton lines, the proportion of neonates on leaf discs of the two lines was not significantly different, but there was significantly higher consumption on CpTI‐Bt transgenic cotton than that on Bt transgenic cotton. In addition, significantly more neonates were found away from the leaf discs, lower consumption and higher mortality were achieved in the choice test with two transgenic cotton leaves than in the choice tests containing non‐transgenic cotton leaves. Leaves and buds were examined in choice tests of fourth instars. It appeared that fourth instars were found in equal numbers on transgenic and non‐transgenic cotton, except when larvae were exposed to leaves for 3 h. However, the total consumption on transgenic cotton was lower than that of the non‐transgenic cotton, so fourth instars may still have the capacity to detect transgenic cotton and reduce feeding on it, although they showed no preference on either transgenic or non‐transgenic cotton. More larvae were found off diet in the treatments with leaves than that of buds, and the number of injured leaf discs by per fourth instar was significantly higher than that of buds in choice tests, suggesting that leaf is a less preferred organ for H. armigera larvae, elicited more larval movements. Similarly, in no‐choice tests of fifth instars, significantly fewer feeding time and more moving time occurred on leaf than that of bud, boll and petal. When cotton line was considered, compared with non‐transgenic cotton, significantly lower feeding time and higher resting time occurred on the two transgenic cottons. Overall, H. armigera larvae have the ability to detect the transgenic Bt and CpTI‐Bt cottons or the less preferred organs and selectively feed more on the non‐transgenic cotton or the preferred organs, especially the neonates, which have a high capacity for avoiding transgenic cotton.     

Degradation of barnacle nauplii: implications to chitin regulation in the marine environment

The exoskeleton of most invertebrate larval forms is made of chitin, which is a linear polysaccharide of β (1→4)-linked N-acetylglucosamine (GlcNAc) residues. These larval forms offer extensive body surface for bacterial attachment and colonization. In nature, degradation of chitin involves a cascade of processes brought about by chitinases produced by specific bacteria in the marine environment. Microbial decomposition of larval carcasses serves as an alternate mechanism for nutrient regeneration, elemental cycling and microbial production. The present study was undertaken to assess the influence of chitinase enzyme on the degradation of the nauplii of barnacle, Balanus amphitrite. The survival and abundance of bacteria during the degradation process under different experimental conditions was monitored. To the best of our knowledge, no such study is conducted to understand the degradation of larval exoskeleton using chitinase and its influence on bacteria. An increase in the chitinase activity with increase in temperature was observed. Scanning electron micrographs of chitinase treated nauplii showed scars on the surface of the barnacle nauplii initially and further disruption of the exoskeleton was observed with the increase in the treatment time. Bacterial abundance of the chitinase treated nauplii increased with the increase in enzyme concentration. Pathogenic bacteria such as Vibrio cholerae, V. alginolyticus, V. parahaemolyticus which were initially associated with the exoskeleton were absent after chitinase treatment, however, Bacillus spp. dominated subsequent to chitinase treatment and this might have important implications to marine ecosystem functioning.     

Synergistic effect and field control efficacy of the binary mixture of permethrin and chlorpyrifos to brown planthopper (Nilaparvata lugens)

The brown Planthopper (BPH), Nilaparvata lugens is a major pest of rice production in tropical Asia. The appearance of insecticide resistance challenges the control of BPH in field. The development of new insecticide is expensive and time-consuming. Thus, the precise and proper use of existing compounds becomes an important issue in resistance management of this pest. In this study, five commercial insecticides of BPH (permethrin, chlorpyrifos, imidacloprid, clothianidin and thiamethoxam) were tested to explore the toxicity of the binary mixture between different kinds of insecticides. In all combinations of mixture, the mixtures of permethrin and chlorpyrifos displays synergistic effect at three different mixture ratios (1:1, 1:10 and 10:1). The strongest synergism observed in permethrin/ chlorpyrifos mixtures at 1:1 ratio (Combination index, CI = 0.39). Addition of enzyme inhibitor followed by detoxification enzyme activity assays suggested that the mechanism of synergistic effect of permethrin/chlorpyrifos mixture may result from inhibition of the cytochrome P450 monooxygenase and esterase activity. This inference can be supported through two lines of evidence. One is decrease of toxicity when permethrin/chlorpyrifos mixture in a 1:1 ratio plus triphenyl phosphate (TPP) or piperonyl butoxide (PBO), but increase of toxicity when permethrin/chlorpyrifos mixture in a 10:1 ratio plus TPP or PBO. Another is exposure of the 3rd instar nymphs to permethrin/chlorpyrifos mixture after 72 h also significantly decreased both cytochrome P450 monooxygenases and esterase activity. Further field trail showed the mixture of 50 ppm permethrin +50 ppm chlorpyrifos increased the field control efficiency significantly rather than permethrin alone or chlorpyrifos alone. Our study indicated that the mixture of permethrin and chlorpyrifos in a 1:1 ratio might be an effective method for the control of BPH in paddy field.