江苏省大中型沼气工程调查及沼液生物学特性研究

近年来,中国大中型沼气工程发展迅速,然而有关沼气工程运行情况的研究甚少。为探索沼气工程运行中存在的问题,该文对江苏省21家畜禽养殖场大中型沼气工程进行了实地调查,并采集发酵料液以及出料样品,分析了进出料液COD(化学需氧量)质量浓度、沼液产气潜力、粪大肠菌群数等指标。结果表明:江苏省沼气工程设计施工规范,配套设施较完备,但运行效率低,沼气、沼液处理或利用能力低。大多数沼气工程以处理养殖废水为主,发酵料液固体质量分数<3%,62%的出料沼液的COD质量浓度达到5000mg/L以上;沼液残余产气潜力较大,在35℃条件下,有12家沼气工程的沼液残余产甲烷量达100mL/L以上。沼气发酵处理可以显著降低粪大肠菌群含量,平均可减少92.9%,但厌氧消化后的沼液中仍含有较高浓度粪大肠菌群,不能达到无害化要求。该调查结果可为畜禽养殖场沼气工程的健康稳定运行与管理提供科学依据。     

Transport Of Fecal Bacteria From Poultry Litter and Cattle Manures Applied to Pastureland

Land applications of manure from confined animal systems and direct deposit by grazing animals are both major sources of bacteria in streams. An understanding of the overland transport mechanisms from land applied waste is needed to improve design of best management practices (BMPs) and modeling of nonpoint source (NPS) pollution. Plots were established on pasturelands receiving phosphorus-based livestock waste applications to measure the concentrations of Escherichia coli (E. coli), fecal coliform (FC), and Enterococcus present in overland flow at the edge of the field. The flow-weighted bacteria concentrations were highest in runoff samples from the plots treated with cowpies (1.37×10⁵ colony forming units (cfu)/100 ml of E. coli) followed by liquid dairy manure (1.84×10⁴ cfu/100 ml of E. coli) and turkey litter (1.29×10⁴ cfu/100 ml of E. coli). The temporal distribution of fecal bacterial concentrations appeared to be dependent upon both the animal waste treatment and the indicator species, with peak concentrations occurring either at the beginning of the runoff event or during peak flow rates. BMPs could be selected to reduce peak flows or first flush effects depending upon the litter or manure applied to the land. The commercial Biolog System was used to identify the dominant species of Enterococcus present in the cowpie source manure (Enterococcus mundtii 55%) and in the runoff collected from the transport plots treated with cowpies (Enterococcus faecalis 37%). The identification of predominant species of Enterococcus that are associated with specific sources of fecal pollution could greatly assist with identifying the origins of NPS pollution.     

Bacterial Transport from Agricultural Lands Fertilized with Animal Manure

A plot scale study was conducted to determine bacterial transport in runoff from cropland treated with poultry litter and dairy manure applied at phosphorus (P) agronomic rates. Treatments included surface application of dairy manure, surface application of poultry litter, incorporation of dairy manure and control. A rainfall simulator was used to induce runoff 1 and 2 days after manure application. Runoff was analyzed to determine the concentration of indicator bacteria-fecal coliform, Escherichia coli, and Enterococcus. Observed edge-of-field bacterial concentrations were 10² to 10⁵ times higher than Virginia’s in-stream bacteria criteria for primary contact recreation waters. No significant treatment effects were observed on edge-of-field bacteria concentration or yield. Results suggest that the manure application based on agronomic P rates may yield significant bacterial loading to downstream waterbodies if rainfall occurs soon after manure application. This research underscores the need for BMPs that reduce runoff volumes and filter pollutants associated with animal manures.     

Identification of an Assemblage of Indicator Organisms to Assess Timing and Source of Bacterial Contamination in Groundwater

Bacterial contamination of drinking water wells is a commonproblem in many rural areas. Some of this contamination may berelated to manure spreading or housing of livestock; another source is on-site septic systems. Current indicator organisms are able to detect the presence of fecal contamination, but where there may be more than one potential source of fecal material, the current indicators are unable to ascertain the origin. This laboratory investigation was undertaken to determine the longevity and reliability of a selected suite of indicator organisms. Total coliform, fecal coliform, fecal streptococci and Clostridium perfringens were monitoredin a simulated contaminated groundwater environment for 6 months. All four indicator organisms were present at the end of6 months. The number of fecal streptococi bacteria decreased most noticeably, allowing assessment of relative age of contamination. C. perfringens was found to be a reliableindicator of contamination from animal manure. Fecal material from 28 different animals and three septic systems were assessed for the presence of the indicator organisms. Totalcoliform, fecal coliform and fecal streptococci were present in the fecal material of all animals tested including reptiles.C. perfringens was detected in feces from all but two of the animals assessed. Using an assemblage of indicator organisms provides more information regarding source and timingof contamination than just testing for total coliform and fecalcoliform bacteria.     

养殖粪水长期贮存过程理化特性变化规律

目前中国中小规模畜禽养殖场主要采用自然贮存后还田的形式处理养殖粪水,受场地制约,养殖粪水贮存时间通常仅有1～2个月,之后便直接还田利用,贮存后的粪水理化特性变化尚不清楚,是否适宜直接还田尚需研究。该研究以猪粪水和牛粪水为研究对象,重点分析粪水在长期贮存中粪大肠菌群、电导率(Electrical Conductance,EC)以及化学需氧量(Chemical Oxygen Demand,COD)的变化,分析粪水最佳贮存期及还田利用方式,以期为粪水资源化及安全还田提供参考。结果表明,粪水经自然贮存6个月,铵态氮损失达68%以上,不仅引起环境污染,且降低了养分;贮存后粪水基本可达到无害化要求,但pH值、EC值以及COD浓度仍然偏高,还田前应制定合理的粪水资源化利用方案;固液分离可以有效降低粪水中的COD浓度和EC值,促进粪水无害化进程。该研究为中国畜禽养殖粪水资源化用探索了新的技术路径。     

Effects of digestate from anaerobically digested cattle slurry and plant materials on soil microbial community and emission of CO2 and N2O

Anaerobic digestion of animal manure and crop residues may be employed to produce biogas as a climate-neutral source of energy and to recycle plant nutrients as fertilizers. However, especially organic farmers are concerned that fertilizing with the digestates may impact the soil microbiota and fertility because they contain more mineral nitrogen (N) and less organic carbon (C) than the non-digested input materials (e.g. raw animal slurry or fresh plant residues). Hence, an incubation study was performed where (1) water, (2) raw cattle slurry, (3) anaerobically digested cattle slurry/maize, (4) anaerobically digested cattle slurry/grass-clover, or (5) fresh grass-clover was applied to soil at arable realistic rates. Experimental unites were sequentially sampled destructively after 1, 3 and 9 days of incubation and the soil assayed for content of mineral N, available organic C, emission of CO₂ and N₂O, microbial phospholipid fatty acids (biomass and community composition) and catabolic response profiling (functional diversity). Fertilizing with the anaerobically digested materials increased the soil concentration of NO₃⁻ ca. 30–40% compared to when raw cattle slurry was applied. Grass-clover contributed with four times more readily degradable organic C than the other materials, causing an increased microbial biomass which depleted the soil for mineral N and probably also O₂. Consequently, grass-clover also caused a ∼10 times increase in emissions of CO₂ and N₂O greenhouse gasses compared to any of the other treatments during the 9 days. Regarding microbial community composition, grass-clover induced the largest changes in microbial diversity measures compared to the controls, where raw cattle slurry and the two anaerobically digested materials (cattle slurry/maize, cattle slurry/grass-clover) only induced minor and transient changes.     

Escherichia coli O157 survival following the surface and sub-surface application of human pathogen contaminated organic waste to soil

Escherichia coli O157:H7 contaminated wastes such as animal manures and abattoir wastes, may be applied directly onto vegetation, the soil surface, or injected deep into the soil. Our aim was to determine the influence of method of waste application to land on E. coli O157:H7 survival. Bovine slurry and ovine stomach contents containing E. coli O157:H7 were applied to the surface vegetation or sub-surface injected at 25 cm below the soil surface. E. coli O157 survived but did not proliferate on grassland vegetation for up to 6 weeks and in the underlying soil for 8 weeks. Our results suggest that sub-surface injection of organic wastes into soil may reduce the risk of pathogen persistence in the environment.     

牛粪水酸化贮存过程中氮形态转化的特性研究

近年来,随着畜禽养殖规模化的快速发展,养殖粪水的处理和利用已成为养殖业健康发展的难点和热点,粪水酸化技术是通过向粪水中添加酸化剂以降低氨气排放,减少粪水贮存中氮素损失的技术,目前此技术已经在丹麦等国推广应用,但中国对此技术的研究尚未起步,为探究粪水酸化固持氮素的效果,该研究以硫酸和明矾为粪水酸化剂,以固液分离前后奶牛粪水为处理对象,通过向粪水中添加酸化剂降低粪水pH值至6.0,分析粪水贮存中氨气排放、氮素转化以及粪大肠菌群数等指标,探索粪水酸化贮存过程氮形态转化机理。研究表明:向养殖粪水中添加酸化剂可降低6.3%~11.1%的总氮损失,能够降低粪水贮存初期中氨气的排放,同时有效抑制了奶牛粪水中粪大肠菌群的活性,使其更易达到无害化处理。酸化剂的加入一方面抑制粪水中微生物作用下的有机氮向无机氮素的转化,提高粪水贮存中有机氮的含量,减少铵态氮的产生量,另一方面酸化剂与粪水中的铵态氮结合生成稳定的铵盐,抑制了粪水中铵态氮向氨气转化的化学平衡,降低了粪水中因氨气排放导致的总氮损失,从而达到减少粪水贮存中氮素损失。     

Fecal coliform dispersal by rain splash on slopes

The movement of fecal pathogens from land to surface and ground water are of great interest because of the public health implications. Non-structural best management practices that control the timing, volume, and placement of animal manures are commonly used to limit opportunities for fecal pathogens to enter water bodies. Increased infiltration capacity, water and waste diversions, and vegetated filter strips are used to control fecal pathogen movement in surface runoff. Fecal pathogens transported by rain splash could conceivably bypass physical barriers. The relationship between slope angle and the transport of fecal coliform bacteria by rain splash was studied. It was hypothesized that there would be a significant down slope transport of fecal coliform bacteria by raindrops falling on a bare soil surface inoculated with fecal coliform bacteria. Slopes from 0° to 40.8° were studied. The mean splash distance for fecal coliforms was less than 50 mm in all directions at 0° slope and more than 500 mm in the downslope direction on a 40.8° slope. Maximum splash distances ranged from about 400 mm on the horizontal surface to more than 1900 mm in the downslope direction on the 40.8° slope. Sequential downhill movement of fecal coliform (FC) bacteria by repeated rain splash could transport FC directly to water bodies or areas of saturation excess where they will become entrained in overland runoff. Further studies on raindrop and rainfall characteristics, as well as surface cover and soil characteristics, will be necessary to more fully understand the mechanisms of FC transport on sloping land by rain splash.     

Occurrence and survival of coliform bacteria, Escherichia coli and Salmonella in various manure and compost

pp. 865–874
Occurrence and survival of fecal-contamination indicator bacteria (coliform bacteria, Escherichia coli and Salmonella ) in various manure and compost samples collected from 23 composting facilities mostly in Kyushu were investigated by using selective media. Coliform bacteria were detected on desoxycholate agar from 11 (38%) of 29 product samples (15 cow dung manure, 4 poultry manure, 2 biosolid compost and 8 food waste compost) at a range of 10² to 10⁶ cfu g¹ dry matter. From positive samples, 21 isolates of possible coliform bacteria were purified. Among them, species of coliform bacteria ( E. coli , E. vulneria , Pantoea sp. and Buttiauxella agrestis ) were identified whereas isolates of Serratia marcescens , not coliform bacteria, were also obtained, suggesting that careful observation was necessary to avoid false positive counting due to the presence of a red colony of S. marcescens that resembled coliform bacteria. Isolates of E. coli were tested for slide aggregation with a set of antiserum against pathogenic E. coli serotypes and negative reaction was obtained for all the isolates tested. Direct detection of E. coli on Chromocult coliform agar and Salmonella on MLCB agar resulted in none and 2 (17%) of 12 samples tested, respectively. The fate of fecal-contamination indicator bacteria as above was followed during compost production on 7 cases at 6 compost facilities and 4 patterns were observed: fecal-contamination indicator bacteria 1) decreased and finally disappeared, 2) decreased once but re-growth was occurred on products, 3) decreased to some extent but remained in products, 4) was not detected throughout production. These results suggest that some fecal-contamination indicator bacteria may survive compost production and appropriate temperature control would be significant for hygiene control of manure and compost.     

猪粪沼液防治辣椒疫病机理研究--沼液中铵与腐殖酸的作用

为探明猪粪沼液对作物土传病害的防治效果与机制,设纯化肥处理为对照,采用盆栽试验比较了猪粪沼液(ADP1)、外源添加铵沼液(ADP2)、除铵沼液(ADP3)、除腐殖酸沼液(ADP4)施用对辣椒疫病的防治效果。结果表明,猪粪沼液(ADP1)可以显著降低辣椒疫病的发病率(防效40.8%),去除沼液中的铵或腐殖酸后,对辣椒疫病的防治效果显著下降,其发病率与病原菌对照(PC)无显著差异。定量PCR检测结果表明,ADP2处理土壤疫霉数量最少(2.51×103copies.g 1),ADP3、ADP4沼液处理的辣椒根际疫霉数量较多,分别为8.19×103copies.g 1、1.38×104copies.g 1;土壤中辣椒疫霉菌的数量低于9.54×103copies.g 1时,辣椒疫病发病率与土壤中病原菌数量有较好的对应关系。不同沼液处理对细菌、真菌和放线菌数量的影响不同。病原菌存在下,ADP1处理的细菌、真菌、放线菌数量均最多,ADP2处理的真菌数量最少。孢子萌发试验表明,随着铵和腐殖酸处理浓度的增加,对疫霉菌游动孢子萌发的抑制率均逐渐增大。浓度为500 mg.L 1、1 000 mg.L 1、1 500 mg.L 1NH4+对疫霉菌孢子萌发的抑制率分别为77.6%、81.8%、95.4%。用浓度为25 mg(C).L 1、50 mg(C).L 1、75 mg(C).L 1、150mg(C).L 1的猪粪沼液腐殖酸处理疫霉菌游动孢子6 h,游动孢子萌发率分别比无菌水对照下降了27.8%、54.5%、70.0%、87.5%。研究表明铵和腐殖酸可能是沼液的主要抑菌因子,沼液作为土传病害抑制剂具有一定的应用前景。     

北京大中型沼气工程冬季运行状况及发酵前后物料理化生物特性

为了解北京市大中型沼气工程冬季运行情况及发酵剩余物理化特性,该文实地调查了北京市29座沼气工程冬季运行情况,以调研问卷的形式了解沼气工程运行状况,并采样分析了发酵原料、沼渣和沼液样品的养分含量、粪大肠菌群数、铜、锌、砷、铅等指标。结果表明:北京29座冬季运行的沼气工程中主要以畜禽粪便为原料,其中有20座沼气工程采用猪粪进行厌氧发酵,厌氧消化反应器类型主要是全混式厌氧反应器和升流式厌氧固体反应器,沼气以供周边农户使用为主,沼渣和沼液以农田利用为主,从区域沼气工程运行状况来看,房山区的沼气工程运行状况相对较好,大兴区和顺义区的沼气工程运行状况参差不齐;所调研沼气工程沼渣和沼液的pH值适宜向农田施用,沼渣的EC值相对原料平均下降了56.97%,沼渣有机质和可挥发性固体的质量分数相对原料分别降低了20.99%和27.93%,沼渣的养分质量分数较高,沼液的养分质量浓度较低,沼渣中的粪大肠菌群数和沼液的化学需氧量、粪大肠菌群数整体偏高,有6座沼气工程的砷含量较高。调查结果可为北京市沼气工程冬季运行效果及沼渣沼液资源化利用提供基础数据,促进北京市沼气工程的可持续发展,提高生态和经济效益。     

Fecal Indicator and Pathogen Reduction in Vegetation Microcosms

Municipal wastewater has been applied as a fertilizer to coniferous forest in northern Sweden with the additional aims to purify the wastewater and reduce the treatment costs. This practice has the potential to introduce pathogenic microorganisms from the applied wastewater or sludge to the environment, with a subsequent risk of transmission to exposed humans or animals. Seasonality and climatological conditions, in particular low temperature, humidity and low sun intensity will furthermore affect the persistence of pathogens in the forest in northern Scandinavia and may therefore enhance the survival and thereby the exposure risk. The survival of two pathogens (Salmonella senftenberg and Campylobacter coli) and four fecal indicator organisms; bacteria strains (E. coli and E. fecalis) and bacteriophages (S. typhimurium phage 28 B and ?x174), used here as viral models, was evaluated in microcosms (60 Litre terrariums) with two vegetation regimes (characterized as moss and lichen) at selected temperatures (+3^?C to +27^?C) and artificial light exposure (light and dark). Salmonella persisted in high numbers after 35 days in the vegetation, indicating a potential risk for further transmission. Neither the vegetation regime nor temperature or light exposure as independent factors governed the reduction of the indicator bacteria and the bacteriophages. The persistence of the indicator organisms was generally best supported in the moss vegetation at low temperature in the dark, where the time for a 90% reduction (T₉₀) was in the range from 8 days (E. coli) to 31 days (Salmonella phage). The die-off characteristics found in this study provide baseline values for further validations in the field and verify that the purification and reuse of municipal wastewater in coniferous forest could potentially be responsible for transmission of fecal pathogens to humans and animals.     

Open‐vessel microwave digestion of animal waste samples for multi‐element analysis

Abstract

Wet acid digestion on a heat block or in a closed‐vessel microwave digestion system normally use half a gram to 1 gram of organic samples, but it is difficult to take a small quantity of representative sample from animal waste materials due to their heterogeneity. This study investigated an alternative microwave digestion system to prepare animal manure samples for plant nutrient analysis. Two types of solid animal manure and two reference plant samples were digested with an open‐vessel microwave digestion system and with the conventional nitric/perchloric acid block digestion, and analyzed for macro‐and micronutrients. Results of the open‐vessel microwave digested samples were comparable to those digested by conventional block digester and in high agreement with the certified values of reference materials. The open‐vessel microwave can reduce digestion time from about 4 hours to about 30 minutes. Fresh samples up to 10 grams can be completely digested directly by the open‐vessel microwave without sample drying and grinding. Sample representation of bulk manure should be better when sample size increased from 1 to 10 grams. Eliminating drying and grinding before digestion improves lab efficiency.     

Development of Regression-Based Models to Predict Fecal Bacteria Numbers at Select Sites within the Illinois River Watershed, Arkansas and Oklahoma, USA

The Illinois River Watershed is a multi-facet basin with ecological and economic importance to its local stakeholders in northwest Arkansas and northeast Oklahoma, USA. The numbers, transport and sources of fecal bacteria in streams was identified as a research priority of the USDA NRI Water and Watershed Program in 2006, and the objective of this study was to evaluate the relation between fecal bacteria and other measured physicochemical parameters in water samples collected from selected sites throughout the Illinois River Watershed. An existing database (i.e., National Water Information Systems, NWIS) from the US Geological Survey (USGS) was used in this project. The data obtained includes discharge, pH, temperature, dissolved oxygen, Escherichia coli (E. coli), fecal coliform, and fecal streptococci among several other physic-chemical parameters. A synthetic model, based on multi-regression analysis, was developed to predict fecal bacteria numbers at these selected sites based on available USGS NWIS data, and the multiple regressions were significant at almost every site for all three bacteria groups. However, the physicochemical parameters used in the equations were very different across sites and fecal bacteria groups, suggesting that the development of such predictive models is site and bacteria group specific even within one watershed.     

Inactivation of Escherichia coli in soil by solarization

Abstract

Contamination of agricultural soil by fecal pathogenic bacteria poses a potential risk of infection to humans. For the biosafety control of field soil, soil solarization in an upland field was examined to determine the efficiency of solarization on the inactivation of Escherichia coli inoculated into soil as a model microorganism for human pathogenic bacteria. Soil solarization, carried out by sprinkling water and covering the soil surface with thin plastic sheets, greatly increased the soil temperature. The daily average temperature of the solarized soil was 4–10°C higher than that of the non-solarized soil and fluctuated between 31 and 38°C. The daily highest temperature reached more than 40°C for 8 days in total in the solarized soil during the second and third weeks of the experiment. Escherichia coli in the solarized soil became undetectable (< 0.08 c.f.u. g^?1 dry soil) within 4 weeks as a result, whereas E. coli survived for more than 6 weeks in the non-solarized soil. Soil solarization, however, had little influence on the total direct count and total viable count of bacteria in the soil. These results indicate that soil solarization would be useful for the biosafety control of soil contaminated by human pathogens via immature compost or animal feces.     

S. typhimurium and E. coli O157:H7 retention and transport in agricultural soil during irrigation practices

During animal waste agricultural applications, the major concern is pathogen spreading, which may contaminate surface water and groundwater. Among the pathogenic microorganisms found in animal waste, Salmonella typhimurium and Escherichia coli O157:H7 are of particular concern. When transported in sub‐surface agricultural soil, S. typhimurium and E. coli O157:H7 are captured at the air–water–sediment interfaces through physical interactions. Because in situ colloids contribute to the formation of air–water–sediment, their mobilization affects the transport of S. typhimurium and E. coli O157:H7. The impact of irrigation rates on in situ colloid mobilization and S. typhimurium and E. coli O157:H7 transport was investigated in intact soil columns collected from an agricultural site in Gadsden County of Florida, USA. The columns were irrigated with sterilized nano‐pure deionized water to mobilize the colloids in the soil by stepwise increases in flow rate. For each flow rate, after colloids were mobilized and steady state was reached, S. typhimurium and E. coli O157:H7 were introduced. The cumulative amount of released in situ colloids increased linearly with the irrigation rates (R² = 0.986–0.996) and transport of the bacteria was enhanced after colloid mobilization. Interactions of the bacteria with the sediments and the air‐water interface were characterized: these played an important role in controlling S. typhimurium and E. coli O157:H7 retention in soil.     

江苏省大型沼气工程沼液理化特性分析

集约化畜禽养殖场沼气工程是一项提供清洁能源、减轻环境污染、潜力巨大的生物质能源工程。该研究采集了江苏省21个规模化养猪场和奶牛场内大型沼气工程的沼液样本,进行了氮磷钾等理化特性分析。结果表明,由厌氧反应罐排出的沼液呈中性至微碱性,还原能力较强。猪场和奶牛场沼液中总氮分别集中在400～900 mg/L和200～400 mg/L范围内,其中铵态氮占总氮量的70%以上;总磷分别集中在30～100 mg/L和300 mg/L以上,可溶性磷含量变异较大;总钾分别集中在100～500 mg/L和500 mg/L以上。沼气工程运行时间对沼液理化性状影响显著,运行时间越长系统稳定性越好。氧化塘对沼液中颗粒物、氮素和磷素有显著的削减作用,多级氧化塘处理效果更明显。     

Vadose Zone Microbial Transport Below At-Grade Distribution of Wastewater Effluent

The attenuation of Escherichia coli and total coliform from secondary treated wastewater effluent under two “at-grade” effluent distribution systems was evaluated in a sandy silt vadose zone in a cold climate. The two at-grade distribution lines had different designs and hydraulic loading rates. Effluent transport was examined using chloride as a tracer. Coliform fate was evaluated relative to the chloride using a combination of in situ pore water sampling and destructive soil sampling, combined with the observation of a dye tracer along excavation sidewalls. Although bacteria attenuation in the subsoil appeared to decrease during colder, winter temperatures (likely due to decreased viability and decreased predation), the subsoil provided about a four log reduction in E. coli over 90 cm of vertical transport. Horizontal transport of bacteria (up to 1.5 m from the line) was likely aided by flow on top of a microbial biomat observed at the soil surface. Both the subsurface dye patterns and the E. coli sampling suggested less preferential flow occurred below the lower loading rate design. At-grade distribution of secondary treated wastewater appears to be a viable alternative to conventional distribution fields at sites with similar climate and soils.