铜绿假单胞菌NY3所产表面活性剂对原油降解的影响

铜绿假单胞菌NY3是从石油污染土壤中分离出的一株能快速代谢疏水性化合物的菌种。研究了该菌产表面活性剂及对原油降解的作用。实验表明,在敞开体系中,投加82 mg/L NY3菌产的鼠李糖脂,240 h能使NY3对原油的降解率提高50%。投加甘油使NY3产鼠李糖脂与降解原油同步进行,与投加鼠李糖脂对原油降解的促进作用相近。投加9‰甘油使NY3在168 h对原油的降解率提高43%。NY3菌能同时降解原油中的直链烷烃及菲(Pr)和芘(Ph)等多环芳烃。在敞开体系中用少量甘油使产鼠李糖脂和降解原油同步进行,节约处理费用。研究结果为NY3菌株在露天石油污染环境修复中的应用奠定了基础。     

鼠李糖脂在铜绿假单胞菌NY3代谢烃过程中的生理作用

为了解鼠李糖脂在菌代谢烃过程中所起的具体作用,对NY3菌体生理生化指标进行测定,研究了NY3菌降解烃过程中,鼠李糖脂促进烃降解与细胞生理特性变化规律的相关性。结果表明,投加60、100和140 mg/L鼠李糖脂,能使正十六烷降解率分别提高23.7%、48.3%和32.2%,且产酸明显。鼠李糖脂能加快疏水性有机物的传质速度,与未加鼠李糖脂相比,鼠李糖脂浓度为100和140 mg/L时,菌细胞内积聚的正十六烷量增加12.5%和37.69%。鼠李糖脂存在下,NY3菌细胞内正十六烷吸收是一个积累和消耗相互交替的动态过程。鼠李糖脂使菌体细胞中疏水性脂类的相对含量和细胞表面的负电荷明显增加,但使所生长的细胞内脂多糖含量明显减少,生长过程投加鼠李糖脂浓度为60、100、140和200mg/L时,细胞中脂多糖含量分别减少45.65%、58.13%、45.61%、和59.44%。因此,鼠李糖脂能加快NY3菌对十六烷的降解,同时对代谢过程中菌体的生理生化特性有明显的改变。     

鼠李糖脂对微生物菌剂降解石油的影响

以石油烃降解微生物菌剂和铜绿假单胞菌株A6为对象，考察不同浓度鼠李糖脂对菌剂细胞表面疏水性、原油降解性能和微生物生长的影响，采用GC-MS分析石油中正构烷烃组分的降解情况。结果表明，低浓度鼠李糖脂就可提高菌剂细胞的表面疏水性和原油降解效果。以250mg／L添加组最明显，第7天疏水性达最高，为58．6％，比对照组提高约26．2％；降解第15天原油降解率达71．6％，分别比对照组和TW20组提高16％和13．3％。GC-MS结果显示鼠李糖脂对高碳数烷烃的降解作用大于低碳数烷烃，正二十三烷和正三十三烷的降解率分别较对照提高了21．5％和33．7％。菌剂对奇数碳烷烃的降解效果优于偶数碳烷烃。鼠李糖脂分别使菌剂中细菌、放线菌和霉菌的最大生物量提高了5．7、2．4和1．8倍。鼠李糖脂对微生物细胞疏水性和生物量的提高与石油降解效果正相关。     

鼠李糖脂对不同菌株降解柴油污染物的影响

通过一系列实验分析了鼠李糖脂对柴油污染物生物降解的影响。单菌株柴油降解实验结果表明,在添加生物表面活性剂鼠李糖脂后,各菌株细胞表面疏水性均发生不同程度的增加,并且对柴油的降解率均有所提高。在混合菌的柴油污染物降解实验中,发现当向土壤中添加了200 mg/L鼠李糖脂时,对柴油的降解才有较大的提高;而当添加100 mg/L的鼠李糖脂到水体中时,对柴油的降解就有较大的提高,而当鼠李糖脂浓度提高为200 mg/L时,柴油的降解率却没有进一步明显的提高。这说明鼠李糖脂对柴油降解的影响程度不仅与环境介质有关,还与添加的鼠李糖脂浓度有关。进一步分析表明,添加适当浓度的鼠李糖脂不仅可以提高对柴油的降解率,而且可加速其降解速度,缩短生物修复所需时间。     

铜绿假单胞菌NY3及突变株代谢油污泥同步产胞外聚合物的特性

油污泥属危险性固体废弃物,大量排放已成为石化企业可持续发展的障碍。利用铜绿假单胞菌NY3及突变株,将油污泥转化为生物质,并研究了其特性。研究表明,NY3、NB1D、NB2D及NB12DD均能降解油污泥中烷烃,但NY3、NB1D和NB2D菌对分子量较高烷烃(从C17~C32)的降解率比双突变菌NB12DD分别平均提高19.68%、28.76%和24.04%。铜绿假单胞菌NY3及突变株能直接将油污泥中烃类转化为胞外聚合物。与未加共代谢碳源相比,葡萄糖作为共代谢碳源时(培养液中表观油浓度达15 g·L~(-1)),培养120 h后,NY3、NB1D、NB2D、NB12DD菌胞外聚合物产量分别提高83.3%、79.8%、20.6%和62.9%,且碱性条件有利于各菌株高效降解油转化为生物质,pH为8~9时,其胞外聚合物产量均最高,分别达到8.1、4.34、4.94和7.15 g·L~(-1),野生菌NY3转化碳源为生物质转化率最高,为52.75%。结构和组成分析表明,胞外聚合物主要为糖蛋白,蛋白4.4%,总糖44.1%,蛋白与糖的连接方式为O-糖苷键链接,91.1%大分子粒径分布在33.0~716.9μm范围。     

生物表面活性剂在城市污泥静态强制通风好氧堆肥中的作用

采用静态强制通风好氧堆肥模式对城市剩余污泥进行堆肥降解,并研究了生物表面活性剂鼠李糖脂对堆肥过程的作用。结果表明,堆肥过程中,添加了质量分数为0.015%鼠李糖脂溶液堆制处理和空白对照堆制处理的堆温变化都明显呈现出中温期0～5 d、高温期6～12 d和降温期13～28 d 3个阶段。实验组比空白组的堆体升温快、高温期持续时间长、堆体的含水率高。鼠李糖脂的添加,使实验组的微生物数量高于空白组。添加鼠李糖脂的堆体和空白堆体的种子发芽指数(GI)在堆肥结束时分别为53.70%和50.80%,说明鼠李糖脂促进了堆肥的腐熟,但由于相对浓缩效应,堆肥产品的重金属含量略高于空白堆体。生物表面活性剂的介入促进了堆肥中木质纤维素的初步降解。研究表明,添加鼠李糖脂能够改善堆肥处理的微环境,促进有机质降解和堆肥的腐熟。     

一株铜绿假单胞菌的筛选鉴定及其产鼠李糖脂条件的优化

从重油污染的土壤中分离出菌株GTX1~GTX4,采用溴化十六烷三甲基铵(CTAB)蓝色凝胶培养基和排油圈法对菌株进行初筛,其中菌株GTX4排油圈直径最大且稳定,通过生理生化实验和16SrDNA鉴定,GTX4为铜绿假单胞菌。将GTX4接种于甘油培养基,在33℃、150r/min条件下培养96h后,经亚甲基蓝-氯仿法和薄层层析(TLC)测定产物为鼠李糖脂,蒽酮-硫酸法测定培养液中鼠李糖脂质量浓度为0.741g/L,培养液表面张力29.87mN/m。通过单因素实验确定GTX4培养的最佳碳源和氮源分别为调和油、NaNO_3,由正交实验得到GTX4产鼠李糖脂的优化条件为调和油45.0g/L,NaNO_3 4.0g/L,微量元素溶液3.5mL/L,菌液接种量0.35%(体积分数),在33℃、200r/min下摇床培养96h,培养液中鼠李糖脂质量浓度可达2.256g/L,液相表面张力降至27.39mN/m。     

二鼠李糖脂对白腐菌降解稻草中木质纤维素的影响

通过固态发酵方式采用白腐菌对稻草中木质纤维素进行降解,并研究了生物表面活性剂二鼠李糖脂对该降解过程的影响。结果表明,不同浓度的二鼠李糖脂能在不同程度上提高降解过程水溶性有机碳的含量,添加0.007%和0.021%二鼠李糖脂的实验组最高TOC(total organic carbon)浓度较对照组分别提高了83.6%和54.5%,这有利于黄孢原毛平革菌的生长,且延缓了菌体的衰退。添加临界胶束浓度0.007%和0.021%浓度的二鼠李糖脂可使LiP(lignin peroxidase)酶活分别提高85.7%和41.2%,二鼠李糖脂对MnP(manganese peroxidase)酶活没有显著影响。生物表面活性剂的介入促进了白腐菌对稻草中木质素的降解,添加0.007%二鼠李糖脂可使木质素降解率提高54%。     

油田含油污泥超声脱油的研究

在油田含油污泥的热洗处理中引入超声辐照处理技术，研究考察了超声脱油技术中超声强度，超声辐照时间，含油污泥预热温度，清洗液用量和清洗液回用4个试验操作条件对污泥脱油的影响，认为在优化条件（超声发生器输出电压175 V，辐照时间15 min，预热温度55 ℃，清洗液和含油污泥以质量比8∶1）下，超声脱油技术可以把污泥的干基油含量由0.35 g/g降低到0.14 g/g，同时也证明清洗液可以反复使用。根据简单的经济核算，处理1 t含油污泥可以产出66元的效益。研究表明，使用超声处理可以有效地提高含油污泥的污油脱除效率，明显降低污泥清洗的操作温度。     

鼠李糖脂浓度对铜绿假单胞菌NY3烃代谢过程中细胞生理特性影响作用

为了研究表面活性剂鼠李糖脂(Rha)促进铜绿假单胞菌NY3烃降解与该菌细胞生理特性变化规律的相关性,在实验室可控条件下,研究了投加Rha后对该菌在整个培养过程中生长情况、正十六烷降解率、胞内脂多糖含量、胞内外蛋白含量等生理特性的影响。结果表明,NY3菌对正十六烷的降解率不受限于该鼠李糖脂临界胶束浓度(CMC=35 mg·L~(-1)),即在投加高于CMC浓度时同样可以提高正十六烷的降解率,并增大体系生物量和积累大量酸性物质(多为烃的氧化产物),该酸性物质能被NY3菌利用并快速生长。投加Rha可增加细胞内脂多糖的含量,并与体系中被去除的烃类物质的量成正比;同时能促进胞内蛋白在短时间内增加并趋于稳定,而降解体系中胞外蛋白同样对于正十六烷的降解有积极作用。因此,投加Rha不仅能提高对疏水性有机物的增溶性,同时对代谢过程中菌体的生理生化特性有明显的改变,从而提高NY3菌对烃类物质的降解率。     

Patterns of volatile metabolites and nonvolatile trichothecenes produced by isolates of <Emphasis Type="Italic">Stachybotrys,Fusarium, Trichoderma,Trichothecium</Emphasis> and <Emphasis Type="Italic">Memnoniella</Emphasis>

We reported previously that trichodiene, a volatile trichothecene derivative, was produced by a Stachybotrys isolate, also known to produce highly cytotoxic, non-volatile, macrocyclic trichothecenes (satrotoxins). We investigated the relationship between the production of trichodiene and various non-volatile trichothecenes for several molds. Volatile metabolites were concentrated by adsorption on Tenax TA and analyzed by GC/MS, while non-volatile metabolites were separated by HPLC, derivatized and analyzed by GC/MS. Stachybotrys chartarum isolates producing macrocyclic trichothecenes secreted significantly larger amounts of trichodiene and other sesquiterpenes than isolates which only produced simple trichothecenes. The amounts of secreted trichodiene were relatively small in all cases. With the exception of Memnoniella, which excreted small amounts of sesquiterpenes, the other isolates produced varying amounts of sesquiterpenes, including trichodiene, as well as simple tricothecenes, no detectable trichodiene, but large amounts of griseofulvin derivatives. In Stachybotrys there is apparently a correlation between trichodiene and macrocyclic trichothecene production. In the remaining isolates, there was no simple relationship between trichodiene and non-volatile trichothecene synthesis. Trichodiene is produced in larger amounts by Stachybotrys isolates, which also produce satratoxins, but it will be difficult to utilize this metabolite to detect toxic isolates in buildings due to the relatively small amounts excreted.     

Radionuclides and trace elements in fish collected upstream and downstream of Los Alamos national laboratory and the doses to humans from the consumption of muscle and bone

Abstract

The purpose of this study was to determine radionuclide and trace element concentrations in bottom‐feeding fish (catfish, carp, and suckers) collected from the confluences of some of the major canyons that cross Los Alamos National Laboratory (LANL) lands with the Rio Grande (RG) and the potential radiological doses from the ingestion of these fish. Samples of muscle and bone (and viscera in some cases) were analyzed for ³H, ⁹⁰Sr, ¹³⁷Cs, ^totU, ²³⁸Pu, ^239,240Pu, and ²⁴¹Am and Ag, As, Ba, Be, Cr, Cd, Cu, Hg, Ni, Pb, Sb, Se, and Tl. Most radionuclides, with the exception of ⁹⁰Sr, in the muscle plus bone portions of fish collected from LANL canyons/RG were not significantly (p<0.05) higher from fish collected upstream (San Ildefonso/background) of LANL. Strontium‐90 in fish muscle plus bone tissue significantly (p<0.05) increases in concentration starting from Los Alamos Canyon, the most upstream confluence (fish contained 3.4E‐02 pCi g^‐1 [126E‐02 Bq kg^‐1]), to Frijoles Canyon, the most downstream confluence (fish contained 14E‐02 pCi g^‐1 [518E‐02 Bq kg^‐1]). The differences in ⁹⁰Sr concentrations in fish collected downstream and upstream (background) of LANL, however, were very small. Based on the average concentrations (±2SD) of radionuclides in fish tissue from the four LANL confluences, the committed effective dose equivalent from the ingestion of 46 lb (21 kg) (maximum ingestion rate per person per year) of fish muscle plus bone, after the subtraction of background, was 0.1 ± 0.1 mrem y^‐1 (1.0 ± 1.0 μSv y^‐1), and was far below the International Commission on Radiological Protection (all pathway) permissible dose limit of 100 mrem y^‐1 (1000 μSv y^‐1). Of the trace elements that were found above the limits of detection (Ba, Cu, and Hg) in fish muscle collected from the confluences of canyons that cross LANL and the RG, none were in significantly higher (p<0.05) concentrations than in muscle of fish collected from background locations.     

A History of Dry Cleaners and Sources of Solvent Releases from Dry Cleaning Equipment

Approximately 25, 000-35, 000 dry cleaning facilities currently operate in the U.S. The release of perchloroethylene and other solvents from these establishments represents a major source of soil and groundwater contamination. The manner in which dry cleaning solvents escape from dry cleaning plants is, for all practical purposes, identical for chlorinated and petroleum hydrocarbon solvents and is related to one of the following events: the catastrophic failure of a component of the dry cleaning system, the improper installation, operation or maintenance of the dry cleaning equipment or a combination of all of these causes. Acceptable customs, codes and regulations can also dictate what is authorized for operation of a dry cleaning facility in a particular community, geographic area during a particular time frame. Environmental litigation dealing with the origin of a solvent release from dry cleaners tends to focus on the design and manufacture of dry cleaning industry machines such as washers, washer extractors, tumblers, solvent filters, water separators, stills and spotting boards. A thorough analysis of the daily operations of dry cleaners often reveals that poor maintenance, failure to follow the manufacturer's instructions and the actions of the operator are the most likely causes of soil and groundwater pollution. In order to forensically evaluate the most probable origins of a solvent release and to examine issues regarding liability, a thorough understanding of the history of dry cleaning and a detailed analysis of the operation and maintenance of the dry cleaning equipment are necessary. The discovery of solvent plumes in the vicinity of dry cleaning plants may suggest that the solvent source is the dry cleaning plant; however, the presence of these plumes does not necessarily indicate that the dry cleaning equipment was defectively designed or manufactured. A thorough review of the type of equipment used over the life of the dry cleaning plant and verifiable solvent mileage records frequently indicates that operators of the plant have disposed of solvent and contaminated solids into the municipal sewer or on ground surfaces.     

Radionuclides in deer and elk from Los Alamos national laboratory and the doses to humans from the ingestion of muscle and bone

Abstract

This paper summarizes radionuclide concentrations (³H, ⁹⁰Sr, ¹³⁷Cs, ²³⁸Pu, ^239,240Pu, ²⁴¹Am, and ^totU) in muscle and bone tissue of mule deer (Odocoileus hemionus) and Rocky Mountain elk (Cervus elaphus) collected from Los Alamos National Laboratory (LANL), Los Alamos, New Mexico, lands from 1991 through 1998. Also, the committed effective dose equivalent (CEDE) and the risk of excess cancer fatalities (RECF) to people who ingest muscle and bone from deer and elk collected from LANL lands were estimated. Most radionuclide concentrations in muscle and bone from individual deer (n = 11) and elk (n = 22) collected from LANL lands were either at less than detectable quantities (where the analytical result was smaller than two counting uncertainties) and/or within upper (95%) level background (BG) concentrations. As a group, most radionuclides in muscle and bone of deer and elk from LANL lands were not significantly higher (p<0.10) than in similar tissues from deer (n = 3) and elk (n = 7) collected from BG locations. Also, elk that had been radio collared and tracked for two years and spent an average time of 50% on LANL lands were not significantly different in most radionuclides from road kill elk that have been collected as part of the environmental surveillance program. Overall, the upper (95%) level net CEDEs (the CEDE plus two sigma for each radioisotope minus background) at the most conservative ingestion rate (50 lbs of muscle and 13 lbs of bone) were as follows: deer muscle = 0.22 mrem y^‐1 (2.2 μSv y^‐1), deer bone = 3.8 mrem y^‐1 (38 μSv y^‐1), elk muscle = 0.12 mrem y^‐1 (1.2 μSv y^‐1), and elk bone = 1.7 mrem y^‐1 (17 μSv y^‐1). All CEDEs were far below the International Commission on Radiological Protection guideline of 100 mrem y^‐1 (1000 μSv y^‐1), and the highest muscle plus bone net CEDE corresponded to a RECF of 2E‐06, which is far below the Environmental Protection Agency upper level guideline of 1E‐04.     

Leaching of seven pesticides currently used in cotton crop in Mato Grosso State—Brazil

This study aimed to evaluate the leaching of pesticides and the applicability of the Attenuation Factor (AF) Model to predict their leaching. The leaching of carbofuran, carbendazim, diuron, metolachlor, α and β endosulfan and chlorpyrifos was studied in an Oxisol using a field experiment lysimeter located in Dom Aquino – Mato Grosso. The samples of percolated water were collected by rain event and analyzed. Chemical and physical soil attributes were determined before pesticide application to the plots. The results showed that carbofuran was the pesticide that presented a higher leaching rate in the studied soil, so was the one representing the highest contamination potential. From the total carbofuran applied in the soil surface, around 6 % leached below 50 cm. The other pesticides showed lower mobility in the studied soil. The calculated values to AF were 7.06E-12 (carbendazim), 5.08E-03 (carbofuran), 3.12E-17 (diuron), 6.66E-345 (α-endosulfan), 1.47E-162 (β-endosulfan), 1.50E-06 (metolachlor), 3.51E-155 (chlorpyrifos). AF Model was useful to classify the pesticides' potential for contamination; however, that model underestimated pesticide leaching.     

Endosulfan in China 2—emissions and residues

Background, aim, and scope  Endosulfan is one of the organochlorine pesticides (OCPs) and also a candidate to be included in a group of new persistent organic pollutants (UNEP 2007). The first national endosulfan usage inventories in China with 1/4° longitude by 1/6° latitude resolution has been reported in an accompanying paper. In the second part of the paper, we compiled the gridded historical emissions and soil residues of endosulfan in China from the usage inventories. Based on the residue/emission data, gridded concentrations of endosulfan in Chinese soil and air have been calculated. These inventories will provide valuable data for the further study of endosulfan. Methods  Emission and residue of endosulfan were calculated from endosulfan usage by using a simplified gridded pesticide emission and residue model—SGPERM, which is an integrated modeling system combining mathematical model, database management system, and geographic information system. By using the emission and residue inventories, annual air and soil concentrations of endosulfan in each cell were determined. Results and discussion  Historical gridded emission and residue inventories of α- and β-endosulfan in agricultural soil in China with 1/4° longitude by 1/6° latitude resolution have been created. Total emissions were around 10,800 t, with α-endosulfan at 7,400 t and β-endosulfan at 3,400 t from 1994 to 2004. The highest residues were 140 t for α-endosulfan and 390 t for β-endosulfan, and the lowest residues were 0.7 t for α-endosulfan and 170 t for β-endosulfan in 2004 in Chinese agricultural soil where endosulfan was applied. Based on the emission and residue inventories, concentrations of α- and β-endosulfan in Chinese air and agricultural surface soil were also calculated for each grid cell. We have estimated annual averaged air concentrations and the annual minimum and maximum soil concentrations across China. The real concentrations will be different from season to season. Although our model does not consider the transport of the insecticide in the atmosphere, which could be very important in some areas during some special time, the estimated concentrations of endosulfan in Chinese air and soil derived from the endosulfan emission and residue inventories are in general consistent with the published monitoring data. Conclusions  To our knowledge, this work is the first inventory of this kind for endosulfan published on a national scale. Concentrations of the chemical in Chinese air and agricultural surface soil were calculated for each grid cell. Results show that the estimated concentrations of endosulfan in Chinese air and soil agree reasonably well with the monitoring data in general. Recommendations and perspectives  The gridded endosulfan emission/residue inventories and also the air and soil concentration inventories created in this study will be updated upon availability of new information, including usage and monitoring data. The establishment of these inventories for the OCP is important for both scientific communities and policy makers.     

Impact of spiked concentrations of Cd,Pb, As and Zn in growth medium on elemental uptake of Nasturtium officinale (Watercress)

This study is aimed at investigating the impact of water quality on the uptake and distribution of three non-essential and toxic elements, namely, As, Cd and Pb in the watercress plant to assess for metal toxicity. The plant was hydroponically cultivated under greenhouse conditions, with the growth medium being spiked with varying concentrations of As, Cd and Pb. Plants that were harvested weekly for elemental analysis showed physiological and morphological symptoms of toxicity on exposure to high concentrations of Cd and Pb. Plants exposed to high concentrations of As did not survive and the threshold for As uptake in watercress was established at 5 ppm. Translocation factors were low in all cases as the toxic elements accumulated more in the roots of the plant than the edible leaves. The impact of Zn on the uptake of toxic elements was also evaluated and Zn was found to have an antagonistic effect on uptake of both Cd and Pb with no notable effect on uptake of As. The findings indicate that phytotoxicity or death of the watercress plant would prevent it from being a route of human exposure to high concentrations of As, Cd and Pb in the environment.     

Butyltin compounds in sediment and fish from the Polish Coast of the Baltic Sea

Concentrations of mono- (MBT), di- (DBT), and tri-(TBT) butyltin compounds were measured in eggs, liver, and muscle of nine species of fish from four regions of the Baltic Sea - the Firth of Vistula, the Gulf of Gdańsk, Puck Bay, and the mouth of the Vistula River. The overall concentration ranges among all the fish sampled from the four sites were: < 7 to 79 ng/g for MBT, 6 to 1100 ng/g for DBT, 7 to 3600 ng/g for TBT, and 16 to 4800 ng/g for total BTs, on a wet wt basis. The highest concentration of total BTs was found in herring liver from the Firth of Vistula (4800 ng/g, wet wt) and in roach muscle from Puck Bay (3300 ng/g, wet wt), while the least concentration was found in burbot eggs and liver from the Vistula River (39 and 32 ng/g, wet wt, respectively). TBT was the major form of BTs present in most samples analyzed. Sediment samples collected from shipyards in the Gulf of Gdańsk contained butyltin concentrations ranging from 1.2 to 46 μg/g (dry wt) for MBT, 2.0 to 42 μg/g for DBT, and 2.6 to 40 μg/g for TBT. As with the fish, the majority of the BTs in sediment were present as TBT, which suggested recent exposure of the aquatic environment of the region to TBT.     

Malathion-induced sublethal toxicity on the hematology of cricket frog (Fejervarya limnocharis)

The effect of malathion [diethyl(dimethoxythiophosphorylthio)succinate] at sublethal concentration (0.006 ppm) on hematological parameters of the cricket frog (Fejervarya limnocharis) was studied for 24 hrs to 240 hrs of exposure and remarkable hematological alterations were observed. The study on hematological parameters revealed a highly significant decrease (P < 0.01) in the total erythrocytes count in malathion-exposed animals from 24 hours to 96 hrs of exposure as compared to control. Significant decreases (P < 0.01) of hemoglobin and packed cell volume were also observed from 48 hrs to 240 hrs. A significant increase (P < 0.01) in leucocytes count was noted throughout the exposure period. Elevated numbers of lymphocytes and eosinophils as found in the present study revealed lymphocytosis as well as eosinophilia, suggesting that this was a result of direct stimulation of the immunological defense due to the presence of a toxic substance or may be associated with tissue damage. The cytomorphological and cytopathological study of erythrocytes and leucocytes in malathion-exposed frogs at 0.006 ppm concentration revealed various cytotoxic effects at different exposure times. It was noted that the size and the shape of the erythrocytes were subjected to variation in different blood disorders.