基于整合生物标志物响应指数评价0号柴油对黑鲷(Sparus macrocephalus)的毒性效应

以0号柴油水溶性成分(water soluble fraction of No.0 fuel oil,WSF)为污染物,采用暴露试验法研究了黑鲷(Sparus macrocephalus)肝脏、鳃和肌肉组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽硫转移酶(GST)、过氧化物酶(POD)活性的变化和脂质氧化水平(MDA)。利用整合生物标志物响应(integrated biomarker responses,IBR,RIB)指数定量化评价0号柴油污染胁迫对黑鲷不同组织的毒性效应。结果表明,黑鲷受0.60、0.30和0.015 mg·L-1WSF胁迫后,肝脏、鳃和肌肉中SOD、CAT和GST酶活性呈现诱导-抑制的规律性变化,POD活性呈现抑制-诱导的规律性变化,MDA含量则呈现先降低后升高的变化趋势。各组织表现出一定的差异性,且肝脏较早诱导酶活性。依据RIB值大小分析得到各组织氧化应激能力水平由高到低依次为肝胰腺、鳃和肌肉。RIB值总体表现为前期升高后期下降的变化趋势,肝脏组织RIB值与WSF浓度具有较好的正相关关系。

基于综合生物标志物响应指数评价0#柴油和平湖原油胁迫下的缢蛏（Sinonovacula constricta）毒性效应

选择0#柴油和平湖原油乳化液对缢蛏(Sinonovacula constricta)进行氧化胁迫实验,选取典型的抗氧化酶-超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷光甘肽硫转移酶(GST)及过氧化物酶(POD)用于衡量油类污染物对生物体造成的氧化压力大小.此外,结合综合生物标志物响应(Integrated Biomarker Responses,IBR)指标,对2种石油污染物对缢蛏的毒性响应进行定量化评价.结果表明,不同浓度的0#柴油和平湖原油对缢蛏消化腺中的4种酶表现出不同程度的诱导效应,各试验组在暴露前期均表现出诱导或抑制,但对4种酶的影响存在时间顺序性,SOD、CAT和GST的酶活性表现为升高-降低的过程,POD表现为降低-升高的过程,活性达到峰值的时间SOD和CAT要早于GST和POD.结合计算出的IBR数值来看,高浓度0#柴油能够引起最为显著的生物效应变化,显示该石油污染物高毒性的特征,0#柴油生物毒性大于平湖原油生物毒性.

Three tropical seagrasses as potential bio-indicators to trace metals in Xincun Bay,Hainan Island,South China

Concentrations of the trace metals Cu, Cd, Pb, and Zn were measured in seawater, rhizosphere sediments, interstitial water, and the tissues of three tropical species of seagrasses （Thalassia hemprichii, Enhalus acoroides and Cymodocea rotundata） from Xincun Bay of Hainan Island, South China. We analyzed different environmental compartments and the highest concentrations of Pb and Zn were found in the interstitial and seawater. The concentrations of Cd and Zn were significantly higher in blades compared with roots or rhizomes in 7＂. hemprichii and E. acoroides, respectively. A metal pollution index （MPI） demonstrated that sediment, interstitial water, and seagrasses in the sites located nearest anthropogenic sources of pollution had the most abundant metal concentrations. There was obvious seasonal variation of these metals in the three seagrasses with higher concentrations of Cu, Pb and Zn in January and Cd in July. Furthermore, the relationships between metal concentrations in seagrasses and environmental compartments were positively correlated significantly. The bioconcentration factors （BCF） demonstrated that Cd from the tissues of the three seagrasses might be absorbed from the sediment by the roots. However, for C. rotundata, Zn is likely to be derived from the seawater through its blades. Therefore, the blades of T. hemprichii, E. acoroides and C. rotundata are potential bio-indicators to Cd content in sediment, and additionally Zn content （C. rotundata only） in seawater.

Leaf Temperature as Thermal Bio-indicator of Heavy Metal Pollutants

Enviromnental pollution is serious social concern. The inflow of heavy metals in the ecological food chain and their subsequent bio-magnification in human bodies is cascading its harmful effects. The metabolism of plants is being hampered by these heavy metals. In the present studies, effect of heavy metal especially cadmium has been studied on stomatal diffusive resistance （SDR）, transpiration rate, leaf surface humidity and leaf temperature of soybean plants growing in hydroponic cultures. Cadmium treatment decelerates the rate of transpiration. The inhibition of transpiration associated with increased SDR leads to dehydration of leaf surface, thereby, increasing temperature of leaves in comparison to control plants. In the present study, temperature difference between stressed and non-stressed plant leaves was as high as 3℃ Moreover, any such rise in leaf temperature due to uptake and induced toxicity of heavy metals can possibly be a good bio-indicator having wide applications in thermal remote sensing and geospatial monitoring of metal polluted soils.

An integrated biomarker response index for the mussel Mytilus edulis based on laboratory exposure to anthracene and field transplantation experiments

Organic pollution is a serious environmental problem in coastal areas and it is important to establish quantitative methods for monitoring this pollution. This study screened a series of sensitive biomarkers to construct an integrated biomarker response （IBR） index using Mytilus edulis. Mussels were exposed to the polycyclic aromatic hydrocarbon anthracene trader controlled laboratory conditions and the activities of components of the glutathione antioxidant system, and the concentrations of oxidative-damage markers, were measured in the gills and digestive glands. Anthracene exposure resulted in increased levels of malondialdehyde （MDA） and superoxide radicals （O2-·）, indicating that oxidative damage had occurred. Correspondingly, anthracene exposure induced increased activities of glutathione S-transferase （GST）, glutathione peroxidase （GPx） and reduced glutathione （GSH） in digestive glands, and GPx and glutathione reductase （GR） in gills, consistent with stimulation of the antioxidant system. A field experiment was set up, in which mussels from a relatively clean area were transplanted to a contaminated site. One month later, the activities of GST, GPx and GR had increased in several tissues, particularly in the digestive glands. Based on the laboratory experiment, an IBR, which showed a positive relationship with anthracene exposure, was constructed. The IBR is suggested to be a potentially useful tool for assessing anthracene pollution.

Comparative Study of the Growth and Gonad Development of Clarias gariepinus （Burchell 1822） Fed Diets with Plant and Animal-based Ingredients in Concrete Tanks

A twelve week experiment was conducted to compare the effects of diets with plant-based and animal-based ingredients on growth and gonad development of Clarias gariepinus. One hundred and 12 sub-adult C. gariepinus with an average weight of 205 ×5.09 g, were stocked in six concrete tanks （9 × 4 × 2 m3） containing 20 fish each. Experiment had 3 replicates and animals fed daily at 5% body weights. There was no significant difference （P 〈 0.05） in proximate compositions of the two experimental feeds and controls. Fecundity, gonad weight and gonasomatic index were higher in fish fed diet 13 than A and C. Similarly, growth indices were higher in fish fed diet B than in A and C. Histology of gonads showed a faster development of oocytes of eggs in fish fed animal-based ingredients than plant-based and combined plant and animal diets. Although there were slight differences in growth parameters and gonad development in favor of feed with animal-based ingredients, plant-based feed compared favorably in the growth and gonad development of C. gariepinus. Plant-based ingredients are recommended on the basis of affordability and availability as substitute for animal-based ingredients in C. gariepinus feed.

Research and Evaluationon Ecotourism Resources of Jilong Valleyin Everest National Nature Reserve

Jilong is a place behind The Himalayas,which is almost isolated from the world.There is typical vertical ecosystem thanks to its special geological environment. So it is honored as “the most abundant species gene warehouse in the Plateau Area”and “the most beautiful valley in the world”.To exploit this valleyis necessary along with the construction of Jilong Porton the border of Sino-Nepal. Based on the research on the spot,the ecological resources there are evaluated and an idea that regards hiking tourism asmain form and ecotourismas the theme is put forward in this paper,which explores a way to develop the valley on the basis of sustainability.

Vegetation-based bioindication of humus forms in coniferous mountain forests

Humus forms, especially the occurrence and the thickness of the horizon of humified residues （OH）, provide valuable information on site conditions. In mountain forest soils, humus forms show a high spatial variability and data on their spatial patterns is often scarce. Our aim was to test the applicability of various vegetation features as proxy for OH thickness. Subalpine coniferous forests dominated by Picea abies （L.） H. Karst. and Larix decidua Mill. were studied in the Province of Trento, Italian Alps, between ca. 900 and 22o0 m a.s.1. Braun-Blanquet vegetation relevds and OH thickness were recorded at 152 plots. The vegetation parameters, tested for their suitability as indicators of OH thickness, encompassed mean Landolt indicator values of the herb layer （both unweighted and cover-weighted means） as well as parameters of vegetation structure （cover values of plant species groups） calculated from the releves. To our knowledge, the predictive power of Landolt indicator values （LIVs） for humus forms had not been tested before. Correlations between OHthickness and mean LIVs were strongest for the soil reaction value, but indicator values for humus, nutrients, temperature and light were also significantly correlated with OH thickness. Generally, weighting with species cover reduced the indicator quality of mean LIVs for OH thickness. The strongest relationships between OH thickness and vegetation structure existed in the following indicators： the cover of forbs （excluding graminoids and ferns） and the cover of Erieaeeae in the herb layer. Regression models predicting OH thickness based on vegetation structure had almost as much predictive power as models based on LIVs. We conclude that LIVs analysis can produce fairly reliable information regarding the thickness of the OH horizon and, thus, the humus form. If no releve data are readily available, a field estimation of the cover values of certain easily distinguishable herb layer species groups is much faster than a vegetation survey with consecutive indicator value analysis, and might be a feasible way of quickly indicating the humus form.

Effect of Carbonation on Microbiologic Parameters of Refrigerated Bovine Raw Milk

This study evaluated the effect of carbon dioxide addition on microbiological quality during refrigerated storage of raw milk collected in Curitiba city, Brazil. A three factor-two level full factorial design was used to investigate the effect of the pH （5.8-6.4）, storage time （0-10 days） and storage temperature （5-10 ℃）, on the responses, namely, mesophiles, psychrotrophs, lipolytic psychrotrophs and proteolytic psychrotrophs counts. Results showed that increase in pH and storage time had significant effect on the microbial count. No significant effect of storage temperature was observed for all the microorganisms studied. All responses were well predicted by selected mathematical models, as denoted by coefficient of determination above 0.95.

The Study of Pollution of PAHs （Polycyclic Aromatic Hydrocarbons） in AI-Hilla River, Iraq by Using Bioindicator Freshwater Crab （Sesarma boulengeri Caiman）

The present study was used freshwater Crab （Sesarma boulengeri Caiman） as bioindicator to measured the PAHs levels in AI-Hilla River, Iraq, during the period from March 2010 to February 2011. Crab samples were collected from two sites on the AI-Hilla River. Sixteen of PAils compounds were identified seasonally in these samples. High mean value was （72.06 μg/g）, recorded by Dibenzo （a, h） anthracene （DbA） in Site 1 and low mean value was （0.36 μg/g） recorded by Anthracene （Ant） in Site 2. The study showed a clear variation in PAHs concentration independent from the seasons and locations. Site 1 recorded high value of HMW （high molecular weight） and LMW （low molecular weight） more than Site 2. The present study revealed that all PAHs compounds were from Pyrogenic Origin. Seven carcinogenic PAHs （cPAHs） were recorded in crab tissues that were collected from the AI-Hilla River. The TEF （toxicity equivalency factor） was calculated to toxicity equivalent crab concentration. From the results, the fresh water Crab Sesarma boulengeri is a good bioindicator to water pollution by PAHs.

The Diversity of the Environmental Impact in Kurzeme

The analysis of environmental impact effects on forest ecosystems has a theoretical and practical nature. Many methods have been developed to determine characteristics and intensity of this impact. Methodologically, they can be divided into three groups： environmental parameter, bioindicative and combined methods. To evaluate the environmental impact a combined method was used in this study, it was based on trees as the bioindicators and their response reactions, and environmental parameter analysis. In this study the bioindicator was Norway spruce （Picea abies （L.） Karst.）, whose response reaction was used to explain total impact volumes of environmental factors in different places in the region of Kurzeme and try to interpret the causes of these differences. As the bioindicators criterion was used response reaction of Norway spruce during a period of 20 years, which was expressed with cumulative and annual additional volume increment, and was depending on the location of the stand and its morphometric characteristics. The empirical material was collected in Kurzeme region in 28 sample plots that are located on two transects and in one reference stand. For the evaluation the widths of the last 40 year growth rings were measured. To express the environmental impact in Kurzeme region a multiple regression model was developed, which explains the environmental impact in the volume of 68.2%, the rest part can be explained by the local conditions of each stand. It must be noted that methodology used in this study is very sensitive, thus, each of the nuances in the dynamics of volume＇s annual reduced additional increments has biological and ecological cause.

Biodiversity in the Republic of Macedonia

Status and trends of biodiversity components in Republic of Macedonia are presented in this article. The monitoring of biodiversity components is conducted according to the criteria of SEBI 2010 （Streamlining European 2010 Biodiversity Indicators） for establishment of consistency at global, regional, EU and national indicator level. Two core segments of the Biodiversity Strategy and Action Plan of Republic of Macedonia are presented as well as the draft list of biodiversity indicators. Ecosystems and biotopes （presented with plant communities） are explained from which 30 Corine habitat classes are identified for the Republic of Macedonia. The number of endemic and endangered species from different taxonomic groups of vascular plants, fungi, fauna, and Emerald species with European importance are presented. Agro-biodiversity, structure of agriculture land, autochthonic breeds of domestic animals as well as the network of protected areas in Republic of Macedonia are presented. At the end conclusion remarks are given for future necessary activities at national and international level regarding monitoring and conservation of the biodiversity.

超高效液相色谱-四极杆/静电场轨道阱高分辨质谱联用同时检测人体尿液中苯、甲苯和二甲苯的6种代谢产物

目的建立同时检测人体尿液中苯、甲苯和二甲苯的6种代谢产物的超高效液相色谱-四极杆/静电场轨道阱高分辨质谱联用方法。方法尿液样品经固相萃取柱净化后,在最佳色谱条件下分离,采用Targeted-SIM/dd-MS2负离子模式扫描,一级精确母离子和二级碎片离子对样品快速定性筛查、一级精确母离子外标法定量分析。优化了色谱柱、流动相、柱温等色谱条件和样品前处理方法及质谱扫描模式等实验参数;测定了方法的检出限、准确度及加标回收等指标;用建立的方法检测了28家涉苯系物企业接触苯系物职工班后尿液中6种代谢产物。结果人体尿液中苯、甲苯、二甲苯的6种代谢产物尿中反-反式粘糠酸(tt-MA)、苯巯基尿酸(SPMA)、马尿酸(HA)、2-甲基马尿酸(2-MHA)、3-甲基马尿酸(3-MHA)和4-甲基马尿酸(4-MHA)在1.0~500μg/L(SPMA:0.1~50μg/L)范围内均有良好的线性关系,相关系数均大于0.999 8,检出限为0.001~0.050μg/L。实际样品的平均加标回收率在90.4%~104.4%范围内,相对标准偏差均低于4.2%。28家涉苯系物企业所采集的155份尿液样品中6种待测物均未超出职业接触限值。结论该方法前处理简单、灵敏度高、检出限低,可有效降低样品假阳性的可能性,适用于接触苯、甲苯和二甲苯生物样本监测指标的同时检测。

血液微小RNAs用于阿尔茨海默病的早期诊断研究

阿尔茨海默病(Alzheimer′s disease,AD)是一种神经系统退行性疾病,主要以进行性记忆损害、认知功能下降、行为改变以及语言障碍为主要临床表现,是老年痴呆中最常见的类型,主要病理改变是老年斑和神经元纤维缠结的形成。目前,对AD尚无有效治愈方法,仅可通过药物延缓其发展过程,因此迫切需要提高AD早期诊断水平,以尽早干预和治疗。微小RNA(micro RNAs)是一种高度保守的小片段非编码RNA,在转录后水平调控基因表达。在神经元形成、分化、突触塑造等过程中起重要调节作用,当神经元发生病变、凋亡时,micro RNAs表达谱将发生改变,研究micro RNAs表达谱变化规律可了解AD发病过程。同时,micro RNAs还可通过血脑屏障,且其二级茎环结构相对稳定不易被降解,这使micro RNAs有望用于AD早期诊断。现将近年micro RNAs表达谱在AD患者脑组织和血液中异常表达研究进展予以综述。

不同倍性虹鳟幼鱼对急性温度胁迫的抗氧化响应

虹鳟属于冷水性鱼类,其生存的最适温度为12～18℃,温度胁迫是虹鳟在夏秋季养殖过程中经常遇到的问题.枫叶鲑和硬头鳟均为品质优良的虹鳟选育种.为探讨急性温度胁迫对两种倍性虹鳟抗氧化响应的影响,本研究选取二倍体枫叶鲑和三倍体硬头鳟幼鱼,分别在13、17、21和25℃下进行热应激试验.在达到目标温度后的0、1、6和12 h取样,之后将受试鱼恢复至13℃的适宜温度下培养,并在恢复培养的、1、12、24和48 h后取样,测定受试鱼肝脏中超氧化物歧化酶(SOD)过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)的活性,以及脂质过氧化物(LPO)、丙二醛(MDA)和热激蛋白70(HSP70)含量.结果表明:枫叶鲑和硬头鳟的3种抗氧化酶活性在17℃组没有出现显著升高;21℃组枫叶鲑和硬头鳟的SOD活性在热应激期间出现显著升高,但枫叶鲑的SOD活性在恢复过程中恢复到正常水平;25℃组枫叶鲑和硬头鳟的SOD、CAT和GPx的活性均显著提高;但在恢复试验进行、24 h后,CAT和GPx的活性恢复到正常水平.枫叶鲑在1721和25℃组HSP70的产生量显著高于13℃组,而硬头鳟仅在21和25℃组HSP70的产生量显著高于13℃组.通过整合生物标志物响应指标值对多种抗氧化参数进行分析,发现枫叶鲑在17和21℃组急性温度胁迫下的抗氧化响应显著高于硬头鳟,但在25℃组硬头鳟的抗氧化响应高于枫叶鲑.这表明不同倍性虹鳟幼鱼在不同温度急性胁迫下的抗氧化响应有所差异.

Effects of heat stress on serum insulin, adipokines, AMP-activated protein kinase, and heat shock signal molecules in dairy cows

Heat stress affects feed intake, milk production, and endocrine status in dairy cows. The temperaturehumidity index（THI） is employed as an index to evaluate the degree of heat stress in dairy cows. However, it is difficult to ascertain whether THI is the most appropriate measurement of heat stress in dairy cows. This experiment was conducted to investigate the effects of heat stress on serum insulin, adipokines（leptin and adiponectin）, AMP-activated protein kinase（AMPK）, and heat shock signal molecules（heat shock transcription factor（HSF） and heat shock proteins（HSP）） in dairy cows and to research biomarkers to be used for better understanding the meaning of THI as a bioclimatic index. To achieve these objectives, two experiments were performed. The first experiment： eighteen lactating Holstein dairy cows were used. The treatments were： heat stress（HS, THI average=81.7, n=9） and cooling（CL, THI average=53.4, n=9）. Samples of HS were obtained on August 16, 2013, and samples of CL were collected on April 7, 2014 in natural conditions. The second experiment： HS treatment cows（n=9） from the first experiment were fed for 8 weeks from August 16, 2013 to October 12, 2013. Samples for moderate heat stress, mild heat stress, and no heat stress were obtained, respectively, according to the physical alterations of the THI. Results showed that heat stress significantly increased the serum adiponectin, AMPK, HSF, HSP27, HSP70, and HSP90（P〈0.05）. Adiponectin is strongly associated with AMPK. The increases of adiponectin and AMPK may be one of the mechanisms to maintain homeostasis in heat-stressed dairy cows. When heat stress treatment lasted 8 weeks, a higher expression of HSF and HSP70 was observed under moderate heat stress. Serum HSF and HSP70 are sensitive and accurate in heat stress and they could be potential indicators of animal response to heat stress. We recommend serum HSF and HSP70 as meaningful biomarkers to supplement the THI and evaluate moderate heat stress in dairy cows in the future.

Effect of scraping therapy on weightlifting ability

OBJECTIVE:To verify the effects of scraping therapy on the weightlifting ability by measuring the subjective sensation,and changes of biomarkers.METHODS:Five students,who have been trained for 3 years in a sport school in China were participated in this study.A course of scraping therapy was applied to intervene during the normal 7-week of weightlifting training programme.The ability of weightlifting,the scale of rating perceived exertion and serum biochemical markers were measured before and after the intervention.RESULTS:Scraping therapy caused a significant increase in weightlifting ability(P<0.05).The level of rating perceived exertion remained stable with the increase in the training volume.Immuno-globulinA was significantly increased(P<0.05),and creatine kinase and blood urea nitrogen were significantly decreased(P<0.05).No significant changes were observed in white blood cell,neutrophil,and testosterone.CONCLUSION:Scraping therapy may facilitate weightlifting ability mainly by decreasing weight sensation and improving serum biochemical parameters.

Bioavailability of Soil Copper from Different Sources:Integrating Chemical Approaches with Biological Indicators

Bioavailability is a key parameter in assessing contaminant transfer to biota. However, the input patterns and soil use types may impact the metal bioavailability. Several soil parameters were measured including chemical properties, such as pH, organic C, and Cu solution/solid speciation, and biological properties, such as soil microbial biomass C （SMBC）, seed germination, and root elongation, to evaluate the bioavailability of Cu contaminated soils from three different sources, i.e., non-ferrous metal mining, Cu-based fungicides, and Cu-smelting. The results revealed that free Cu2＋ ion in soil solution and the ratios of Cu fractions to total Cu content in the solid phase could not be used to predict total Cu content in soils. The indexes of seed germination and root elongation appeared not to be good biomonitors of Cu contamination in soils, which were more sensitive to soil pH and soil organic carbon （SOC）. Relationships between SMBC and soil Cu forms or the ratio of SMBC/SOC and soil Cu forms showed that free Cu2＋ ion and humie acid-complexed Cu could significantly inhibit soil microbial activities. Our findings suggested that both metal chemical forms and biological bioassays should be considered as a complementary technique rather than an alternative to evaluate the metal bioavailability from different pollution sources.

Genotoxicity biomarkers in aquatic bioindicators

Pollution of the aquatic environment is an ever-growing problem, as waters are the ultimate sink for the large number of xenobiotics from multiple sources. DNA damaging agents have a significant ecological relevance since they are implicated in many pathological processes and exert effects beyond that of individual being active through following generations. A large number of methods have been applied to evaluate genotoxic damage in different aquatic species. Comet assay, as method for de- tecting DNA alterations, and micronucleus test, as an index of chromosomal damage are the most widely applied and validated methods in field studies. These methods were applied in different vertebrate and invertebrate aquatic species, but only mollusk and fish species have been employed in routine biomonitoring programs. Mussels, due to their widely geographical distribution and the suitability for caging represent the bioindicator of choice in field studies. Mytilus species is the most used marine mussel. The use of fish is limited to specific geographic areas. The present review mainly focuses on the application of comet assay and micronucleus test in mussels. A number of biomonitoring studies in mussels, using comet assay or micronucleus test, revealed exposure to different classes of genotoxic compounds with a good discrimination power. The different evidence from the two as- says, reflects different biological mechanisms for the two genetic endpoints, DNA damage and chromosomal damage, suggesting their combined application in the field. Different endogenous and exogenous factors have been shown to modulate the genotoxic responses in mussels, acting as confounding factors in environmental monitoring. The use of standardized protocol for caging, sampling and genotoxity evaluation is critical in biomonitoring studies. The use of a multimarker approach coupling genotoxicity biomarkers with physiological and biochemical factors allows to have a complete picture of the environmental pollution [Current Zoology 60 （2）： 273-284, 2014].

EQUIVALENT UNIFORM DOSE SENSITIVITY TO CHANGES IN ABSORBED DOSE DISTRIBUTION

Treatment planning in external beam radiation therapy （EBRT） utilizes dose volume histograms （DVHs） as optimization and evaluation tools. They present the fraction of planning target volume （PTV） receiving more than a given absorbed dose, against the absorbed dose values, and a number of radiobiological indices can be computed with their help. Equivalent uniform dose （EUD） is the absorbed dose that, uniformly imparted, would yield the same biological effect on a tumor as the dose distribution described by the DVH. Uncertainty and missing information can affect the dose distribution, therefore DVHs can be modeled as samples from a set of possible outcomes. This work studies the sensitivity of the EUD index when a small change in absorbed dose distribution takes place. EUD is treated as a functional on the set of DVHs. Defining a Levy distance on this set and using a suitable expansion of the functional, a very simple expression for a bound on the variation of EUD when the dose distribution changes is found. This bound is easily interpreted in terms of standard treatment planning practice.