转化异丁香酚生成香草醛纺锤芽孢杆菌的筛选

以底物异丁香酚为唯一碳源从七壤中筛选获得了一株能高效转化异丁香酚生成香草醛的芽孢杆菌。根据生理生化特性及16SrRNA序列分析鉴定其属于纺锤芽孢杆菌（Bacillus fusiformis）,初步试验表明该菌能转化2％异丁香酚生成4．20g/L香草醚。     

短短小芽孢杆菌启动子筛选载体的构建

为了筛选短短小芽孢杆菌强启动子元件,应用PCR技术从枯草杆菌168中分离出α-淀粉酶基因,用其作为报告基因与质粒pUB110和pKF3一起构建了启动子筛选载体pKB/A.将短短小芽孢杆菌细胞壁蛋白基因启动子引入该载体构建成重组质粒pKB/PA,电穿孔法转化短短小芽孢杆菌50后发现α-淀粉酶以活性形式分泌表达.结果表明短短小芽孢杆菌启动子筛选载体构建成功.     

转化异丁香酚生成香草醛纺锤芽孢杆菌的筛选*

以底物异丁香酚为唯一碳源从土壤中筛选获得了一株能高效转化异丁香酚生成香草醛的芽孢杆菌。根据生理生化特性及16S rRNA序列分析鉴定其属于纺锤芽孢杆菌(Bacil-lus fusiformis),初步试验表明该菌能转化2%异丁香酚生成4.20 g/L香草醛。     

一株凝结芽孢杆菌的分离筛选及产孢条件优化

【背景】凝结芽孢杆菌除了具有一般乳酸菌的益生功能外,还具较强的耐酸、耐胆盐、耐高温、易贮存等生物特性。【目的】从泡菜中筛选一株性能优良的凝结芽孢杆菌用于微生态制剂的制备,并对其产孢率进行优化,为该菌株的进一步工业化生产提供参考依据。【方法】采用选择性培养基通过特定的培养条件,筛选到一株抑菌效果良好的产酸芽孢杆菌,并对其进行特异性引物的鉴定、16S rRNA基因序列分析及生理生化实验。通过单因素及正交试验对菌株的产芽孢条件进行优化。【结果】筛选得到一株凝结芽孢杆菌BC01,该菌株对大肠杆菌(Escherichia coli CVCC 1527)、鼠伤寒沙门氏菌(Salmonella typhimurium CVCC 2228)、产气荚膜梭菌(Clostridium perfringens CVCC 46)、猪霍乱沙门氏菌(Salmonella choleraesuis CVCC 503)等均有较强的抑制作用;模拟胃液处理120 min存活率达到94%;0.3%的胆盐存活率达到84.3%。单因素及正交试验优化后的最适培养基配方:糖蜜10.0 g/L,酵母浸出粉20.0 g/L,NaCl 5.0 g/L,K_2HPO_4 5.0 g/L,MnSO_4 10.0 mg/L;最适培养条件:接种量4%,温度45°C,初始pH 7.0,转速200 r/min,培养时间36 h。在该优化条件下,其活菌数最高达到6.7×10~9 CFU/mL,产孢率达到89.2%。【结论】筛选得到一株可用于微生态制剂的菌株——凝结芽孢杆菌BC01,对其产孢率进行了优化,为工业化生产奠定了基础。     

我国森林土壤中苏云金芽孢杆菌生态分布的研究

从我国8个森林立地带(寒温带、中温带、暖温带、北亚热带、中亚热带、南亚热带、高原亚热带、热带)所属的13个自然保护区,采集了0—5cm土层林下土壤样品384个.测定了土壤pH、水分和养分.从中分离观察芽孢杆菌菌落1873个,分离出苏云金芽孢杆菌79株,并对其所属亚种进行了初步鉴定.其平均出土率和分离率分别为14.32%和4.21%.研究了芽孢杆菌和苏云金芽孢杆菌在森林土壤中生态分布的规律及苏云金芽孢杆菌对6种昆虫的室内毒力测定,从中筛选出不少的高效菌株.为研究苏云金芽孢杆菌在我国森林生态系中资源的保护、开发和利用,具有重要意义.     

具有分泌蛋白能力的短芽孢杆菌的筛选及鉴定

短芽孢杆菌（Bacillus　brevis）具有分泌蛋白能力强和胞外蛋白酶活性低的特性,是分泌表达外源蛋白较理想的宿主。为获得分泌表达系统较理想的宿主菌,建立了短芽孢杆菌高效筛选模型,从800余株细菌中筛得8株具有高蛋白分泌能力且没有胞外蛋白酶活性的候选菌。经多相分类学初步鉴定其中5株为短芽孢杆菌。     

产高活性抗凝溶栓双活性蛋白的短小芽孢杆菌的筛选及鉴定

【目的】筛选可产生抗血栓活性物质的细菌。【方法】利用VY/4平板、酪蛋白平板从水样、土样、兔粪、羊粪、朽木等20多个样品中筛选目的菌株;利用纤维蛋白平板和纤维蛋白试管检测抗血栓活性;利用形态学特征、理化性质、16S rRNA序列同源性鉴定目的菌株。【结果】得到5株可产生抗血栓活性物质的细菌,重点研究了菌株LDS33,发现其分泌的胞外蛋白在纤维蛋白平板上和纤维蛋白试管中均显示出强烈的溶栓活性,通过试管法发现此蛋白质同时具有较强的抗凝活性。结合形态学、理化性质、16S rDNA序列及进化树分析,发现该菌株属于硬壁菌门芽孢杆菌目芽孢杆菌科芽孢杆菌属的短小芽孢杆菌,将其命名为Bacillus pumilus LDS.33。【结论】短小芽孢杆菌LDS33可产生高活性的抗凝溶栓双活性蛋白。     

茄子黄萎病菌抗性根际芽孢杆菌的筛选与鉴定

利用离体抑菌圈法从茄子根际土壤筛选出8株对黄萎病菌大丽轮枝菌(Verticillium dahliae)抑菌效果明显的芽孢杆菌,其中F53菌株分泌几丁质酶和-β1,3-葡聚糖酶,对所检测的12种植物病原菌具明显的抑制作用,其发酵液热、酸碱稳定性强,用F53菌株制成的菌剂对茄子黄萎病盆栽试验防效为52.4%~75.8%,初步鉴定F53菌株为环状芽孢杆菌(Bacillus circulans)。     

药用芽孢杆菌对常用中草药的耐药性试验

目的 研究药用芽孢杆菌对几种常用中草药的耐药性,为药用芽孢杆菌与中草药的临床合并应用提供理论依据.方法 将中草药制成煎剂,通过琼脂扩散方法进行药敏试验.结果 枯草芽孢杆菌(Bacillus subtilis)、蜡样芽孢杆菌(Bacillus cereus)和纳豆芽孢杆菌(Bccillus natto)3种芽孢杆菌除了对黄连和黄芩有一定敏感性外,对其余8种中草药均不敏感;而地衣芽孢杆菌(Bacillus clicheniformis)则对黄连、黄芩和金银花都有一定敏感性,对其余中草药均不敏感.结论 4种药用芽孢杆菌对大部分中草药(包括有杀菌和抑菌作用的中草药)具有耐药性.     

亚麻微生物脱胶菌种的筛选与鉴定

在研究天然水沤法脱胶的过程中,通过初筛、复筛,从沤麻主生物期的沤麻液中筛选出两株茵落周围产生透明圈较大、脱胶酶活较高的菌株。通过形态观察,并对其多项生理、生化指标进行了分析研究,初步鉴定并命名为枯草芽孢杆菌A1和B1。初步加茵脱胶实验表明：枯草芽孢杆菌A1产生果胶酶、木聚糖酶,而不产生纤维素酶,脱胶周期为72小时;枯草芽孢杆茵B1产生果胶酶、木聚糖酶和纤维素酶,脱胶周期为50小时。     

Sublethal effects of deltamethrin exposure of parental generations on physiological traits and overwintering in Leptinotarsa decemlineata

Although the evolution of insecticide resistance has received a lot of attention, less is known about sublethal effects of insecticide stress experienced by the preceding generations on the performance of pest populations. We investigated whether three generations of parental exposure to a deltamethrin insecticide influences physiological traits and overwintering success of offspring in Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae). Beetles descending from insecticide‐exposed parents had lower adult body mass but higher relative lipid content and resting metabolic rate than those descending from non‐insecticide‐exposed parents. Also, a higher proportion of beetles descending from insecticide‐exposed parents than from control parents overwintered on the soil surface, which was associated with very low overwintering survival. When burrowed into the soil for overwintering, both groups had similar probability to survive the overwintering period. Parental insecticide exposure can reduce overall overwintering survival in the next generation by disturbing overwintering behaviour. Although beetles descending from insecticide‐exposed parents were small, the overwintering conditions seem to select for high lipid content, which could override the negative effects of small weight, and possibly improve fitness in the following growing season. Thus, insecticide application may have unintended consequences, which should be considered in pest management.     

Insecticide resistance in the dengue vector Aedes aegypti from Martinique: distribution, mechanisms and relations with environmental factors

Dengue is an important mosquito borne viral disease in Martinique Island (French West Indies). The viruses responsible for dengue are transmitted by Aedes aegypti, an indoor day-biting mosquito. The most effective proven method for disease prevention has been by vector control by various chemical or biological means. Unfortunately insecticide resistance has already been observed on the Island and recently showed to significantly reduce the efficacy of vector control interventions. In this study, we investigated the distribution of resistance and the underlying mechanisms in nine Ae. aegypti populations. Statistical multifactorial approach was used to investigate the correlations between insecticide resistance levels, associated mechanisms and environmental factors characterizing the mosquito populations. Bioassays revealed high levels of resistance to temephos and deltamethrin and susceptibility to Bti in the 9 populations tested. Biochemical assays showed elevated detoxification enzyme activities of monooxygenases, carboxylesterases and glutathione S-tranferases in most of the populations. Molecular screening for common insecticide target-site mutations, revealed the presence of the "knock-down resistance" V1016I Kdr mutation at high frequency (>87%). Real time quantitative RT-PCR showed the potential involvement of several candidate detoxification genes in insecticide resistance. Principal Component Analysis (PCA) performed with variables characterizing Ae. aegypti from Martinique permitted to underline potential links existing between resistance distribution and other variables such as agriculture practices, vector control interventions and urbanization. Insecticide resistance is widespread but not homogeneously distributed across Martinique. The influence of environmental and operational factors on the evolution of the resistance and mechanisms are discussed.     

Molecular surveillance of the antifolate-resistant mutation I164L in imported african isolates of Plasmodium falciparum in Europe: sentinel data from TropNetEurop

Background

Malaria control programmes utilising indoor residual spraying are only effective if a high coverage of targeted structures is achieved and an insecticide that is effective against the specific mosquito vector is correctly applied. Ongoing monitoring of spraying operations is essential to assure optimal programme performance and early corrective action, where indicated.

Methods

Successful development and application of a computerised spraying operations management system in Mpumalanga Province, South Africa during 1998 resulted in its adaptation and introduction in neighbouring Maputo Province, southern Mozambique during 2000. The structure and components of this computerised management system are described, and its' operational benefit in southern Mozambique, where community-based spray operators apply intradomiciliary insecticide, are reviewed.

Conclusions

The computerised management system allowed malaria programme management and field supervisors to monitor spraying coverage, insecticide consumption and application rates on an ongoing basis. The system supported a successful transition to community-based spraying, while assuring correct insecticide application and spraying completion according to schedule.     

Insecticide susceptibility and detoxication enzyme activities among Coptotermes formosanus Shiraki workers sampled from different locations in New Orleans

Coptotermes formosanus Shiraki worker termites were sampled from 20 locations in the City Park area of New Orleans, LA. The termites were subsequently assayed to determine their susceptibility to cypermethrin, chlordane and chlorpyrifos, and detoxication enzyme activity. Cypermethrin was most toxic against Formosan subterranean termite workers, chlorpyrifos exhibited intermediate toxicity and chlordane was least toxic. A comparison of insecticide susceptibility between the most and least tolerant colonies revealed 1.9-, 1.7- and 1.8-fold differences in susceptibility for cypermethrin, chlorpyrifos and chlordane, respectively. As with the bioassay data, although significant differences were noted, a great deal of overlap was observed among the colonies for total cytochrome P450 content (difference of 2.2-fold between high and low value) aldrin epoxidation (3.6-fold) and cytosolic esterase (3.9-fold) activity. No significant differences were observed among the colonies for methoxyresorufin O-demethylase or glutathione S-transferase activity. Conversely, microsomal esterase activity varied greatly; a 38-fold difference was observed between the most (Cf1776) and least (Cf1387) active colonies. However, no significant correlation was observed between insecticide susceptibility and microsomal esterase activity. In fact, no significant correlations were observed between any of the enzyme activities measured and insecticide susceptibility. These results are discussed in the context of insecticide selection and future control effectiveness.     

The fate, distribution and biological performance of insecticide residues in vegetable crops following seedling treatment

Carrot, cauliflower and radish seedlings raised in sand culture and treated with aqueous suspensions containing chlorfenvinphos, diazinon, iodofenphos or triazophos were planted into the field in 1982 and 1983 to compare the performance against cabbage root fly and carrot fly of these treatments with that of bow-wave applications of granular formulations at sowing. Initial residue concentrations were up to 4100 mg kg^-1 in the seedlings at planting. Thereafter the amounts of insecticides per root system declined. Residue concentrations in mature carrot and radish roots were smaller when plants had been treated as seedlings rather than by bow-wave application. Chlorfenvinphos was the most effective insecticide against both pests. Amounts of insecticide applied in transplanted seedlings were < 5% of the amounts applied by the bow-wave method but short-term control of cabbage root fly on radish was similar. Carrot fly damage was reduced by 60% by some of the seedling treatments after 16 wk in 1982 but in 1983 they were less effective. Pot experiments with chlorfenvinphos showed that most of the insecticide in the seedlings was transferred rapidly into the surrounding soil and it was concluded that the bioactivity resulted mainly from uniform distribution of the released insecticide between and around individual plants.     

Implications of gene amplification for evolution and management of insecticide resistance

Simulations were used to compare evolution of insecticide resistance predicted by a conventional two-allele model with predictions from three- and four-allele models that assume resistance is based on gene amplification. Results were similar between models when insecticide concentration was low or moderate. In contrast, when 10% of the population was not exposed to insecticide each generation, high insecticide concentrations slowed resistance development in the two-allele model, but caused rapid development of high levels of resistance in the three- and four-allele models. The presence of a third allele at an initial frequency as low as 10-7 doubled or tripled the rate of resistance development in some cases. Attempts to slow evolution of resistance by overwhelming it with high concentrations of insecticides are not likely to succeed if gene amplification or other mechanisms generate alleles that confer high levels of resistance.     

Evaluating reproductive fitness and metabolic costs for insecticide resistance in Myzus persicae from Chile

The development of insecticide resistance in pest insects is an increasing problem for agriculture, forestry and public health. Aphids are ubiquitous herbivorous insects, with approximately 4700 known species, of which less than 5% exploit the agricultural environment successfully. Of these, the peach‐potato aphid Myzus persicae Sulzer is recognized as one of the most important pests worldwide because it has acquired resistance to many insecticides. Although resistance to insecticides provides important benefits for pests in agricultural fields that are treated with insecticides, it may be associated with fitness (or other) costs in environments that are insecticide free. In the present study, the fitness and energy costs that might be experienced by M. persicae in an insecticide‐free environment when carrying at least one insecticide resistance mutation (IRM), or by having an increased production of esterases, are evaluated. The study investigates whether genotypes that have an IRM also have enhanced esterase production, whether there is any metabolic cost associated with insecticide resistance, and whether there are any fitness costs associated with insecticide resistance and metabolic expenditure. The intrinsic rate of increase, standard metabolic rate (i.e. a measure of maintenance costs) and constitutive esterase activity are determined for 30 different multilocus genotypes carrying (or not carrying) at least one of the two most frequent insecticide resistance mutations (MACE and kdr/super‐kdr) that occur in Chile. The results show that genotypes carrying at least one IRM have higher levels of total esterase activity than genotypes without an IRM, that there is no evidence of an energy cost associated with total esterase activity or IRM, and no evidence for a reproductive fitness cost associated with total esterase activity, IRM or metabolic rate. The results agree with previous studies showing linkage disequilibrium between insecticide resistance mechanisms, although they contrast with those of studies that report fitness costs associated with insecticide resistance in Myzus persicae.     

Pyrethroid resistance in mosquitoes

Repeated blood feedings throughout their life span have made mosquitoes ideal transmitters of a wide variety of disease agents. Vector control is a very important part of the current global strategy for the control of mosquito-associated diseases and insecticide application is the most important component in this effort. Pyrethroids, which account for 25% of the world insecticide market, are currently the most widely used insecticides for the indoor control of mosquitoes and are the only chemical recommended for the treatment of mosquito nets, the main tool for preventing malaria in Africa. However, mosquito-borne diseases are now resurgent, largely because of insecticide resistance that has developed in mosquito vectors and the anti-parasite drug resistance of parasites. This paper reviews our current knowledge of the molecular mechanisms governing metabolic detoxification and the development of target site insensitivity that leads to pyrethroid resistance in mosquitoes.     

Current Perspectives on Plague Vector Control in Madagascar: Susceptibility Status of Xenopsylla cheopis to 12 Insecticides

Plague is a rodent disease transmissible to humans by infected flea bites, and Madagascar is one of the countries with the highest plague incidence in the world. This study reports the susceptibility of the main plague vector Xenopsylla cheopis to 12 different insecticides belonging to 4 insecticide families (carbamates, organophosphates, pyrethroids and organochlorines). Eight populations from different geographical regions of Madagascar previously resistant to deltamethrin were tested with a World Health Organization standard bioassay. Insecticide susceptibility varied amongst populations, but all of them were resistant to six insecticides belonging to pyrethroid and carbamate insecticides (alphacypermethrin, lambdacyhalothrin, etofenprox, deltamethrin, bendiocarb and propoxur). Only one insecticide (dieldrin) was an efficient pulicide for all flea populations. Cross resistances were suspected. This study proposes at least three alternative insecticides (malathion, fenitrothion and cyfluthrin) to replace deltamethrin during plague epidemic responses, but the most efficient insecticide may be different for each population studied. We highlight the importance of continuous insecticide susceptibility surveillance in the areas of high plague risk in Madagascar.     

Effects of selected insecticides on adults of two parasitoid species of Liriomyza trifolii: Ganaspidium nigrimanus (Figitidae) and Neochrysocharis formosa (Eulophidae)

Abstract Liriomyza trifolii is an important pest of vegetables and ornamental crops around the world. This pest is attacked by many parasitoid species. The principal management tactic used against L. trifolii is insecticide application. Insecticides vary in their effects on parasitoid species and insecticides that have less harmful effects should be preferred for the control of this pest. In this study, novaluron, abamectin, λ‐cyhalothrin and spinetoram were investigated for their lethal effects on adults of Neochrysocharis formosa and Ganaspidium nigrimanus, two important parasitoids of L. trifolii. Three different bioassays were used on adult parasitoids: direct insecticide application, insecticide intake and insecticide residue. Adult parasitoid response to novaluron exhibited the least lethal effects among the bioassays and insecticides tested. Abamectin had significant mortality to both parasitoid species in the direct application and insecticide intake bioassays and mortality were high for G. nigrimanus in the residue bioassay. Spinetoram was the most harmful insecticide to the adult parasitoids in all three bioassays. λ‐cyhalothrin effects varied between the two parasitoids. In the direct application, it was harmful to G. nigrimanus and had no effect on N. formosa. In the insecticide intake bioassay λ‐cyhalothrin had no effect in survival of either species, and in the residue bioassay it reduced parasitoid survival of both species. Potential tolerance of N. formosa to λ‐cyhalothrin is discussed.