Cloning of mpd gene from a chlorpyrifos-degrading bacterium and use of this strain in bioremediation of contaminated soil

An effective chlorpyrifos-degrading bacterium (named strain YC-1) was isolated from the sludge of the wastewater treating system of an organophosphorus pesticides manufacturer. Based on the results of phenotypic features, phylogenetic similarity of 16S rRNA gene sequences and BIOLOG test, strain YC-1 was identified as the genus Stenotrophomonas. The isolate utilized chlorpyrifos as the sole source of carbon and phosphorus for its growth and hydrolyzed chlorpyrifos to 3,5,6-trichloro-2-pyridinol. Parathion, methyl parathion, and fenitrothion also could be degraded by strain YC-1 when provided as the sole source of carbon and phosphorus. The gene encoding the organophosphorus hydrolase was cloned using a PCR cloning strategy based on the known methyl parathion degrading (mpd) gene of Plesiomonas sp. M6. Sequence blast result indicated this gene has 99% similar to mpd. The inoculation of strain YC-1 (10(6) cells g(-1)) to soil treated with 100 mg kg(-1) chlorpyrifos resulted in a higher degradation rate than in noninoculated soils. Theses results highlight the potential of this bacterium to be used in the cleanup of contaminated pesticide waste in the environment.     

杀虫剂及不同地区寄主植物对棉蚜酯酶的影响

通过滤纸酯酶反应法比较 4种有机磷杀虫剂对北京和山东高密地区取食不同寄主植物的棉蚜β-乙酸萘酯 (β -NA)酯酶的抑制作用。结果表明高密地区不同寄主植物上的棉蚜 β -NA酯酶活力高的个体多于北京地区 ,同时辛硫磷对高密地区不同寄主植物上的棉蚜 β -NA酯酶的抑制作用亦高于北京地区 ,而久效磷和DDVP对两地区不同寄主植物上的棉蚜种群中 β-NA酯酶的抑制作用相似。     

Malathion applied at standard rates reduces fledgling condition and adult male survival in a wild lesser kestrel population

The lesser kestrel Falco naumanni is a globally threatened colonial bird that has suffered a generalized population decline in Europe. Recent studies have suggested that the land-use changes linked to agriculture intensification are the main factors accounting for the population regression. However, there has been little analysis of the role of pesticide applications. In this study, we examined the consequences of a malathion treatment in a wild population of lesser kestrels, specifically its effects on breeding performance, adult survival and population size. We found that the larger the area treated with malathion around the colony, the lower the size and the body condition of the fledglings, although no effects were found on the number of fledged chicks per pair or their sex ratio. Survival of adult males, but not females, was lower in the malathion-treated areas. These results show that organophosphorus pesticides applied at standard rates might disrupt the lesser kestrel population dynamics by reducing their breeding performance and increasing adult mortality in a sex-biased way. The proportional area treated around the colony did not affect colony size in the following year, indicating that an increased adult male mortality was not enough to lead to a detectable population size reduction and/or that the arrival of immigrants could have masked it. Both the direct malathion toxic effects and the expected reduction in prey availability due to fumigation are likely to underlie the observed effects. Hence, keeping non-treated buffer areas around kestrel colony centers should be a strictly applied conservation measure to avoid the observed negative side effects of malathion applications.     

Inheritance of chorionic malformations and insecticide resistance by Spodoptera exigua

Abstract:  Effect of exposure to the organophosphorus insecticide – fenitrothion [O,O-dimethyl-O-(4-nitro-3-methyl)phenyl] – on four generations of Spodoptera exigua Hübner (Lep., Noctuidae) larvae, with regard to the mortality and structure of the chorion of eggs laid was tested. Generations varied in the total mortality rate, although all of them showed some survival rate. The third and the fourth generations showed the lowest mortality among tested ones. Exposure to fenitrothion caused malformations in eggs. The chorion revealed cracks and diminutions of the outer layer. The quality and quantity of malformations increased proportional to the duration of exposure, e.g. the second exposed generation reveald more prominent changes than the first one. Malformations were observed in the next generations, which were not exposed to pesticide. The follicular cells, which are responsible for the structure and sculpture of the eggshell, must have inherited the malforming mechanism. Most probably, malformation takes place during the late development of eggs in the ovarioles.     

Biomarkers as Predictors in Health and Ecological Risk Assessment

Biomarkers are measurable biological parameters that change in response to xenobiotic exposure and other environmental or physiological stressors, and can be indices of toxicant exposure or effects. If the biomarkers are sufficiently specific and well characterized, they can have great utility in the risk assessment process by providing an indication of the degree of exposure of humans or animals in natural populations to a specific xenobiotic or class of xenobiotics. Most biomarkers are effective as indices of exposure, but adequate information is rarely available on the appropriate dose-response curves to have well-described biomarkers of effect that can be widely applicable to additional populations. Specific examples of acetylcholinest-erase inhibition following exposure to organophosphorus insecticides are cited from experiments in both mammals (rats) and fish. These experiments have indicated that the degree of inhibition can be readily influenced by endogenous (e.g., age) and exogenous (e.g., chemical exposures) factors, and that the degree of inhibition is not readily correlated with toxicological effects. Caution is urged, therefore, in an attempt to utilize biomarkers in the risk assessment process until more complete documentation is available on the specificity, sensitivity, and time course of changes, and on the impact of multiple exposures or the time of exposures.     

Phytoremediation of chlorpyrifos by Populus and Salix

Chlorpyrifos is one of the commonly used organophosphorus insecticides that are implicated in serious environmental and human health problems. To evaluate plant potential for uptake of chlorpyrifos, several plant species of poplar (Populus sp.) and willow (Salix sp.) were investigated. Chlorpyrifos was taken up from nutrient solution by all seven plant species. Significant amounts of chlorpyrifos accumulated in plant tissues, and roots accumulated higher concentrations of chlorpyrifos than did shoots. Chlorpyrifos did not persist in the plant tissues, suggesting further metabolism of chlorpyrifos in plant tissue. To our knowledge, this work represents the first report for phytoremediation of chlorpyrifos using poplar and willow plants.     

The involvement of acetylcholinesterase in resistance of the California red scale shape Aonidiella aurantii to organophosphorus pesticides

Laboratory toxicity bioassays using chlorpyrifos (Dursban) confirmed the notion that development of resistance is responsible for widespread failures to control the California red scale, Aonidiella aurantii (Mask.) by applying organophosphorus (OP) compounds in citrus groves in Israel. Higher Vmax values of acetylcholinesterase (AChE) activity (9–13 fold) were measured in resistant strains collected from the field as compared to a susceptible line. No differences were found with respect to Km values using acetylthiocholine iodide as a substrate, or degree of inhibition (expressed by IC₅₀ values) by the OP compounds chlorpyrifos-oxon and paraoxon and the carbamate pirimicarb. We suggest that resistance of the California red scale is caused by excess of AChE molecules able to bind and thus scavenge inhibitory OP compounds. This scavenging mechanism related to AChE may be similar in other insect species where elevated levels of detoxifying esterases were implicated in conferring OP resistance.     

How do earthworms affect microfloral and faunal community diversity?

Much of the work regarding earthworm effects on other organisms has focused on the functional significance of microbial-earthworm interactions, and little is known on the effects of earthworms on microfloral and faunal diversity. Earthworms can affect soil microflora and fauna populations directly and indirectly by three main mechanisms: (1) comminution, burrowing and casting; (2) grazing; (3) dispersal. These activities change the soil's physico-chemical and biological status and may cause drastic shifts in the density, diversity, structure and activity of microbial and faunal communities within the drilosphere. Certain organisms and species may be enhanced, reduced or not be affected at all depending on their ability to adapt to the particular conditions of different earthworm drilospheres. A large host of factors (including CaCO₃, enzymes, mucus and antimicrobial substances) influence the ability of preferentially or randomly ingested organisms to survive (or not) passage through the earthworm gut, and their resultant capacity to recover and proliferate (or not) in earthworm casts. Small organisms, particularly microflora and microfauna, with limited ability to move within the soil, may benefit from the (comparatively) long ranging movements of earthworms. Microflora and smaller fauna appear to be particularly sensitive to earthworm activities, and priming effects enhancing nutrient release, particularly in casts, are common. Larger fauna such as microarthropods, enchytraeids and Isopods may be enhanced under some conditions (e.g., in earthworm middens), but in other cases earthworm activity may lead to a decrease in their populations due to competition for food (microbes and organic materials), and spatial and temporal changes in food abundance. Nevertheless, considering the presently available data, the beneficial interactions of earthworms and microflora and fauna appear to far outweigh the potential negative effects. However, much is still unknown regarding the interactions of earthworms of different ecological categories on the diversity and function of microfloral and faunal communities, and much more interdisciplinary research is needed to assess the potential role of earthworms in regulating the diversity of microflora and fauna in soil systems and the potentially beneficial or harmful effects this regulation may have on ecosystem function and plant growth in different ecosystems.     

不同种植年限菜田土壤微生物区系的研究

采用稀释平板涂抹法对山东聊城周边地区不同种植年限的菜田土壤微生物数量、组成与养分状况及其相互关系进行了研究。结果表明：①该地区土壤微生物以细菌占绝对优势,其次为放线菌;②随着种菜年限的增加,土壤细菌和微生物总数表现出“低高低”的变化趋势,而真菌和放线菌则随种菜年限的增加而增加;③菜田土壤各类微生物主要集中在0～20cm的土层中,随着土层加深,其数量迅速减少;④菜田土壤放线菌组成复杂,共分离到了6个属的放线菌,但仍以链霉菌为主,其次为小单胞菌属和马杜拉放线菌属。链霉菌可分为9个类群,白孢类群占优势。种植1—2a的菜田土壤放线菌组成复杂,而链霉茼组成较简单;⑤蔬菜种类不同时,土壤微生物亦不同;⑥土壤养分含量与真菌和放线菌呈负相关,与细菌和微生物总数呈正相关。     

韭菜和土壤中毒死蜱的残留与降解动态

在大棚和露地栽培条件下,研究了不同浓度的毒死蜱灌根施药后土壤和韭菜中毒死蜱的残留与降解动态.结果表明:韭菜中毒死蜱的降解速度比土壤中快,平均半衰期分别为3.41 d和7.40 d;在大棚和露地栽培条件下,韭菜中毒死蜱的降解速率差异不大,平均降解半衰期分别为3.37和3.44 d.施药灌根后第21天,韭菜中毒死蜱的残留量(0.021 ～0.102mg·kg-1)基本低于GB 2763-2005规定的最大残留限量标准(≤0.1 mg·kg-1).新生韭菜中仍残留少量毒死蜱,但明显低于药后第一次刈割.土壤中残留的毒死蜱对韭菜中的农药残留量有显著影响.     

种植年限对蔬菜日光温室土壤微生物区系和酶活性的影响

以种植2、4、6、11、13、16、19年的蔬菜日光温室土壤为研究对象,并以露地菜田为对照,测定了土壤微生物区系及酶活性的变化.结果表明: 随着种植年限的增加,土壤中细菌、放线菌和微生物总数均呈现先增加后减少的趋势,在种植11年时达到最大值,分别比对照增加了54.8%、63.7%和55.4%,差异达显著水平;而真菌数量持续上升,种植19年约为对照的2.2倍.微生物生理类群中,纤维素分解菌、自生固氮菌、亚硝酸细菌、反硝化细菌和硫化细菌数量的变化趋势与细菌相似, 种植11年分别为对照的1.5、1.6、1.9、1.4和1.1倍;而氨化细菌数量则呈现先减少后增加的趋势,在种植13年时达到最小值,为对照的56.0%.土壤中脲酶、多酚氧化酶、蔗糖酶、蛋白酶、纤维素酶和碱性磷酸酶活性随种植年限的增加呈现先增强后减弱的趋势,而过氧化氢酶活性较稳定.相关分析表明,细菌、放线菌和微生物总数与各土壤酶均呈显著正相关;而真菌数量与脲酶、蔗糖酶、过氧化氢酶和碱性磷酸酶均呈负相关,其中与过氧化氢酶的相关性达到显著水平.     

Character of interactions of saprophytic soil microflora via gaseous metabolites

The character of interaction between saprophytic soil bacteria via gaseous metabolites was studied. It was established that, at the metabolic level, a diverse character of interspecies interrelationships between bacteria exist, directly influencing their reproduction and preservation in soil. Volatile compounds produced by bacteria are able to act as both intra-and interspecies regulators of microbial communities. The soil microbiocenosis composition may be therefore regulated by volatile products of metabolism of saprophytic soil bacteria. Methanol released by bacteria into the environment plays an important role in this process.     

丹参红叶病发生的微生态机制

通过对丹参红叶病株与健康株根区土养分含量及根区土和根表土中的微生物区系比较,探索丹参红叶病发生的微生态机制.结果表明:丹参红叶病株叶片中N、P、K、Mn含量均显著低于健康株(P＜0.05);根区土中速效P与健康株根区土无显著差异,速效N、K含量均显著高于健康株根区土(P＜0.05),表明丹参红叶病害发生与P缺乏有关,但植株缺磷不是由于土壤供磷不足所致.丹参红叶病株根区土细菌数量较健康株减少41.3％,真菌和放线菌数量分别较健康株增加156.6％和189.5％,差异均达显著水平(P＜0.05);丹参红叶病株根表土细菌、真菌和放线菌数量变化趋势与根区土—致.在丹参红叶病株根区、根表土壤中,6种优势真菌、4种优势放线菌及2种优势细菌可能为有害微生物.优势真菌为腐皮镰刀菌、露湿漆斑菌、三线镰刀菌、焦曲霉、尖孢镰刀菌及座囊菌;优势放线菌为砖红链霉菌、威威达湖伦茨氏菌、马铃薯疮痂病原链霉菌及山丘链霉菌;优势细菌为阿氏芽孢杆菌及水生细菌.这些优势微生物可能通过影响根系生长及根系对土壤养分吸收引起丹参出现缺磷现象.表明丹参红叶病的发生与丹参根区土和根表土中微生物区系异常密切相关.     

川芎及土壤中毒死蜱残留检测提取方法

为建立川芎植株和土壤中农药毒死蜱残留检测的最佳提取方法,利用气相色谱-质谱法测定毒死蜱残留量,并采用正交设计,考察提取溶剂、提取时间、提取次数和溶剂量对农药毒死蜱回收率的影响,优选提取方法。川芎地上部分和根茎中毒死蜱残留的最佳提取方法为：乙腈3倍量超声提取2次,每次15 min。土壤中毒死蜱残留的最佳提取方法为：乙酸乙酯10倍量震荡提取2次,每次30 min。经方法学验证,筛选建立的提取方法均达到农药残留分析要求,该方法可行。     

全球大气CO2浓度升高对土壤微生物的影响

全球大气CO2浓度升高对土壤微生物生态系统的影响已引起广泛关注。本文从土壤微生物群落结构、微生物区系、土壤呼吸、微生物生物量以及土壤酶活性方面对大气高浓度CO2的响应进行了综述。由于提供高浓度CO2的实验系统、所选植物材料以及土壤特性等的不同,大气CO2浓度升高对土壤微生物群落结构、微生物区系、土壤呼吸、微生物生物量以及土壤酶活性的影响并未得出一致结论。但高浓度CO2对土壤微生物生态系统的影响是存在的。     

种植盐地碱蓬改良滨海盐渍土对土壤微生物区系的影响

利用盐生植物盐地碱蓬对天津河口滨海盐碱地进行生物修复,研究了其对土壤微生物区系的影响.结果表明,种植区碱蓬根系土壤的可溶性盐分与对照土壤相比下降了41%（重量法）和37%（电导法）;根系土壤的微生物数量明显增加,其中细菌、放线菌和真菌分别较对照增加了2.3倍、4.3倍和71倍,与对照相比均为显著性差异.根系微生物的耐盐性结果显示,随着土壤盐分的降低,根系微生物生长的最适盐度也随之降低,耐盐性较低的微生物种群已逐渐成为优势种群.系统发育分析表明,枯草杆菌属成为植物修复后土壤中的优势种群.