黑曲霉固态发酵三七渣产淀粉酶的研究

以三七渣为基质,采用黑曲霉固态发酵产淀粉酶,考察了硫酸铵(氮源)添加量、三七渣粒径、固体培养基含水率、发酵温度、发酵时间、菌悬液接种量等因素对产淀粉酶效果的影响,并采用正交实验对发酵条件进行了优化。单因子实验结果表明,黑曲霉固态发酵三七渣产淀粉酶适宜的硫酸铵添加量为40～60mg/g(以干药渣计),最佳三七渣粒径为100目,最适固体培养基含水率为55%,最佳发酵温度为34℃,最适发酵时间为7d,最佳菌悬液接种量为10%(质量分数)。正交实验多重比较的结果表明,优化的发酵条件为:硫酸铵添加量50mg/g,发酵温度34℃,发酵时间5d,在此发酵条件下,黑曲霉固态发酵三七渣产生的淀粉酶的酶活可达84.15U/g(以湿物料计)。     

黑曲霉固态发酵三七渣产纤维素酶

考察了硫酸铵添加量、磷酸二氢钾添加量、三七渣粒径、培养基含水率等因素对黑曲霉固态发酵三七渣产纤维素酶的影响,并采用正交实验对培养基制备条件进行了优化。研究结果表明,三七渣作为发酵培养基用于生产纤维素酶是可行的;含水率、硫酸铵添加量、磷酸二氢钾添加量以及三者之间的交互作用对实验结果的影响极显著(P0.01),这3个因素以及它们之间的交互作用对实验结果影响程度的排序为:硫酸铵添加量含水率磷酸二氢钾添加量交互作用;优化的培养基制备条件为:采用过80目的筛三七渣,硫酸铵添加量为40 mg/g干药渣,磷酸二氢钾添加量为10 mg/g干药渣,初始含水率为55%,pH值自然,在此条件下发酵,所产CMC酶活可达73.23 IU/g湿物料。     

培养基特性对三七渣发酵产红曲色素的影响

考察了蛋白胨添加量、磷酸二氢钾添加量、含水率、培养基初始pH值、基质粒径对紫色红曲霉固态发酵三七渣产红曲色素的影响。结果表明:三七渣固态发酵生产的红曲色素中黄色素的色价显著高于红色素;培养基特性对红色素和黄色素产量的影响规律相似;适宜的发酵培养基特性为:采用过60或80目筛的三七渣,蛋白胨添加量5%~6%,磷酸二氢钾添加量1%,培养基含水率60%~70%,初始pH=5。     

三七渣固态发酵生产绿色木霉的工艺条件研究

考察了发酵时间、孢子液接种量、固态发酵培养基装料量、培养温度、光照条件等对三七渣固态发酵生产绿色木霉的影响,并采用正交实验优化了发酵工艺条件。结果表明,发酵过程受工艺条件的影响较大,孢子液接种量、固态发酵培养基装料量、发酵时间3个因素的交互作用极显著(p0.01),这3个因素以及它们之间的交互作用对实验结果的影响程度排序为:交互作用固态发酵培养基装料量孢子液接种量发酵时间;在最佳的发酵工艺条件(即培养温度24℃、无光照、接种量20%(1g三七渣接种2mL绿色木霉孢子液)、固态发酵培养基装料量12.5g、发酵时间8d)下,平均产孢量可达10.10×109 cfu/g。     

三七渣固态发酵生产康宁木霉生防菌

考察了发酵时间、接种量、装料量、温度、光照条件对三七渣固态发酵生产康宁木霉生防菌的影响,并采用正交实验优化了发酵工艺条件。研究结果表明,将三七渣转化为环境友好的康宁木霉生防菌是可行的。发酵过程受工艺条件的影响较大,发酵时间、接种量、装料量3个因素的交互作用极显著(p0.01),这3个因素以及它们之间的交互作用对实验结果影响程度的排序为:发酵时间交互作用装料量接种量。优化的发酵工艺条件为:发酵温度30℃、无光照、接种量15%、装料量7.5 g、发酵时间6 d,在此条件下,康宁木霉的产孢量可以达到16.75×109cfu/g。     

海带渣废弃物资源化利用以及多功能菌肥固体发酵条件的优化

海带渣是海带加工过程中主要固体废弃物。本实验利用海带渣对生防菌株JGA2(-5)27-2进行固体发酵。实验对不同发酵条件下发酵产物对植物病原真菌寄生曲霉的抑制活性与对发酵产植物激素6-糠氨基嘌呤的能力进行测定。通过单因素实验和响应面实验优化了发酵条件:蔗糖为最优的碳源;硝酸盐为最优的氮源;蔗糖浓度、KNO3浓度、最佳的培养时间、含水量、培养温度、接种量和pH值分别为2.091%、0.507%、8.24 d、60%、35℃、4%和自然pH值。在发酵过程中以添加KNO3的形式向发酵菌肥中加入K+离子。实验证明,利用生防菌株JGA2(-5)27-2进行海带渣的固体发酵生产多功能微生物肥料具有可行性,并为海带渣的资源化在农业领域的利用提供一定参考。     

响应面法优化香蕉秸秆与猪粪混合厌氧发酵工艺研究

将香蕉秸秆与猪粪混合后厌氧发酵,考察粪秸比(猪粪干物质占发酵原料干物质的质量分数)、厌氧污泥接种量(厌氧污泥占发酵料液的质量分数)和发酵温度对总产气量的影响。在此基础上,通过响应面法对厌氧发酵工艺进行优化,并建立总产气量与粪秸比、厌氧污泥接种量、发酵温度的二次回归模型。研究结果表明,建立的二次多项式数学模型具有高度显著性,模型拟合度、精确度高,数据合理。最佳工艺条件为:粪秸比为35.34%,厌氧污泥接种量为61.40%,发酵温度为40.27℃。在此条件下,总产气量的预测值为15 779.2mL,试验值为15 620.5mL,二者相对偏差为1.01%。可见,所得模型能够较好地优化发酵条件并预测总产气量,可为提高香蕉秸秆与猪粪混合厌氧发酵产气量及提高发酵效率提供一定参考。     

氨氧化菌群的筛选及发酵条件的优化

利用选择性培养基对活性污泥进行连续驯化,筛选出氨氮去除效率较高且稳定的氨氧化菌群。采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术分析了氨氧化菌群在连续传代过程中菌群的结构差异,并对菌群的发酵培养基及发酵条件分别进行了正交优化和单因子优化。结果表明,该菌群的最佳发酵培养基为每升培养基中含碳酸盐缓冲液15mmol,硫酸铵4.2 mmol,磷酸盐缓冲液12.5 mmol,硫酸亚铁0.9μmol,氯化钙0.4 mmol,硫酸镁1.5 mmol;最佳发酵条件为:接种量为14%,装液量为60 mL/250 mL,温度为35℃。在此条件下,菌群对氨氮的去除效果较优化前提高了155%。     

调理料添加比例对菇渣基质发酵效果的影响

为探讨废弃蘑菇基料菇渣改性生产有机栽培基质技术,直接采用菇渣为主要原料,以猪粪作为调理料进行好氧发酵,研究猪粪的不同添加比例对菇渣好氧发酵腐熟条件中的温度、挥发性固体(VS)、p H、全氮、全磷、种子发芽指数(GI)的影响。结果表明,猪粪适宜作为菇渣改性生产有机基质的调理料,发酵过程始终维持含水率在65%左右,3个处理高温持续时间均在5 d以上,满足基质无害化卫生标准要求,发酵完成后p H均符合腐熟基质标准,GI均达到60%以上。而猪粪添加量较少时,可以缩短进入高温发酵期的时间,堆体达到的最高温度较高,堆料腐熟较快,且最有利于氮素的保存和全磷的浓缩。综合以上实验结果表明,在菇渣发酵中添加15%的猪粪有利于改善发酵条件,提高堆体的养分含量。     

黄姜纤维素渣固态发酵生产蛋白饲料

利用酵母菌和黑曲霉对黄姜纤维素渣进行固态发酵生产蛋白饲料。研究了接种量、温度、固液比、发酵时间和通风量对发酵的影响。同时在单菌种发酵的基础上,对酵母菌和黑曲霉的混合发酵进行了初步探索,研究结果表明,混菌发酵的实验效果比单菌发酵的效果好。当条件为:黄姜纤维素渣25 g,加入脲0.53 g,KH2PO40.05 g,K2HPO40.05 g,Mg-SO40.05 g,NaCl 0.05 g,CaCl20.01 g,接种量为14%,温度30℃,固液比2∶1,发酵产物的蛋白质量分数可达到13.98%。     

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.