The air-borne distribution of zoonotic agents from livestock waste spreading and microbiological risk to fresh produce from contaminated irrigation sources

Aims:  To assess the risks of zoonotic agents in dissemination of livestock wastes into the environment by airborne distribution. To subsequently assess the survival time of zoonotic agents, introduced in irrigation water, on the phylloplane of produce.
Methods and Results:  An Escherichia coli marker was introduced into pig slurry which was spread using a rain gun sprayer. Air sampling was undertaken to determine the distance reached by the marker. No recoveries were observed at a distance of 250 m. Borehole water, contaminated with zoonotic agents, was used to irrigate field plots sown with lettuce and spinach. Decline in bacterial numbers on the phylloplane was observed with time. After initial rapid decreases, we were unable to detect any pathogen from the phylloplane, 1 month after contamination.
Conclusions:  These preliminary results suggest that the risks to public health from the aerosolized spread of bacteria during slurry spreading by rain gun are low. Although, zoonotic agents on crop phylloplanes perish quickly, the risks of overhead irrigation of fresh produce 3 weeks before harvest should still be considered.
Significance and Impact of the Study:  These preliminary results improve our understanding on the fate of zoonotic agents in the environment. Spreading liquid livestock wastes by an airborne mechanism may not pose a significant public health risk. Detection of zoonotic agents 3 weeks after contamination of lettuce and spinach means that consideration should be given by the farmers until the time of harvest, when irrigating fresh produce with water that may have been directly or indirectly contaminated by livestock wastes.     

Evaluation of soil saturation, soil chemistry, and early spring soil and air temperatures as risk factors in yellow-cedar decline

Yellow‐cedar (Callitropsis nootkatensis (D. Don) Oerst.) is a valuable tree species that is experiencing a widespread decline and mortality in southeast Alaska. This study evaluated the relative importance of several potential risk factors associated with yellow‐cedar decline: soil saturation, soil aluminum (Al) toxicity or calcium (Ca) deficiency, and air and soil temperature. Data were collected from permanent vegetation plots established in two low‐elevation coastal forests exhibiting broad ranges of cedar mortality. Measurements of each risk factor were contrasted among classified forest zones to indicate if there were strong links with decline. Hydrology alone is weakly associated with yellow‐cedar decline, but could have a predisposing role in the decline by creating exposed conditions because of reduced forest productivity. Yellow‐cedar decline is not strongly associated with soil pH and extractable Al and Ca, but there appears to be Ca enrichment of surface soils by feedback from dead yellow‐cedar foliage. Air and soil temperature factors are strongly associated with decline. Based on these results, an hypothesis is presented to explain the mechanism of tree injury where exposure‐driven tree mortality is initiated in gaps created by soil saturation and then expands in gaps created by the tree‐mortality itself. The exposure allows soils to warm in early spring causing premature dehardening in yellow‐cedar trees and subsequent freezing injury during cold events. Yellow‐cedars growing in the protection of shade or snow are not preconditioned by this warming, and thus not as susceptible to cold injury. Yellow‐cedar decline appears to be associated with regional climate changes, but whether the cause of these changes is related to natural or human‐induced climate shifts remains uncertain. Management implications, the possible role of climate, and recommended research are discussed.     

膜下滴灌条件下滴水量和滴水频率对棉田土壤水分分布及水分利用效率的影响

通过两年的田间试验,研究了滴水量和滴水频率对膜下滴灌棉田土壤水分分布及棉花水分利用效率的影响.结果表明:从整个生育期来看,当滴水量(375 mm)相同时,高频滴灌(每3天1次)处理0～20 cm土层含水率较高而深层土壤湿润不够;低频滴灌(每10天1次)处理有利于水分的下渗和侧渗,深层土壤含水率较高,但水分补给不及时,表层土壤偏低;总体上中频滴灌(每7天1次)处理有利于水分在土壤剖面中的均匀分配.当滴水频率相同时,滴水量越大,土壤含水率越高,40 cm以下土层含水率也越高.不同处理的棉田耗水规律基本一致,苗期较低,平均不高于1.7 mm·d-1,蕾期开始上升至花铃期达到最高,日均耗水量可达8.7 mm·d-1,吐絮期回落到1.0 mm·d-1左右.总耗水量与降水和滴水量密切相关,而与滴水频率无关;滴水频率对棉花水分利用效率无显著影响,但水分利用效率随滴水量的增大而显著降低.少量滴灌(300 mm)虽然可以获得较高的水分利用效率,但减产严重,过量滴灌(450mm)无显著增产效应,水分浪费严重.在当地棉田自然条件下,采用中量(375 mm)+中低频(每7天或10天1次)的滴灌模式为宜.     

外科血培养标本中病原菌分布及耐药性分析

目的了解中山大学附属第一医院外科血标本中病原菌的菌种分布及常见菌株的耐药性。方法血标本用Bact/A lert-120全自动血培养仪进行血培养,阳性血培养转种后用VITEK-60 AMS细菌鉴定仪鉴定,用K-B法进行药敏试验。结果2002年1月至2005年12月血培养标本中共分离出病原菌256株,阳性率为10.6%。122株(47.7%)为革兰阴性杆菌,其中肠杆菌科细菌占72.1%(88/122),非发酵菌占27.9%(34/122);113株(44.1%)为革兰阳性球菌,其中葡萄球菌属占51.3%(58/113),肠球菌占38.1%(43/113);真菌21株(8.2%)。血培养中的革兰阴性杆菌对亚胺培南、美洛培南治疗敏感;革兰阳性球菌对万古霉素和替考拉宁敏感。结论肠杆菌科细菌和葡萄球菌是外科血培养中的主要病原菌,其耐药现象严重,宜根据药敏结果选用敏感抗菌药物治疗。     

Inactivation of a human norovirus surrogate by high-pressure processing: effectiveness, mechanism, and potential application in the fresh produce industry

Fresh produce is often a high-risk food for norovirus contamination because it can become contaminated at both preharvest and postharvest stages and it undergoes minimal or no processing. Currently, there is no effective method to eliminate the viruses from fresh produce. This study systematically investigated the effectiveness of high-pressure processing (HPP) on inactivating murine norovirus (MNV-1), a surrogate for human norovirus, in aqueous medium and fresh produce. We demonstrated that MNV-1 was effectively inactivated by HPP. More than a 5-log-PFU/g reduction was achieved in all tested fresh produce when it was pressurized at 400 MPa for 2 min at 4°C. We found that pressure, pH, temperature, and food matrix affected the virus survival in foods. MNV-1 was more effectively inactivated at 4°C than at 20°C in both medium and fresh produce. MNV-1 was also more sensitive to HPP at neutral pH than at acidic pH. We further demonstrated that disruption of viral capsid structure, but not degradation of viral genomic RNA, is the primary mechanism of virus inactivation by HPP. However, HPP does not degrade viral capsid protein, and the pressurized capsid protein was still antigenic. Overall, HPP had a variable effect on the sensorial quality of fresh produce, depending on the pressure level and type of product. Taken together, HPP effectively inactivated a human norovirus surrogate in fresh produce with a minimal impact on food quality and thus can provide a novel intervention for processing fruits intended for frozen storage and related products such as purees, sauces, and juices.     

Microtensiometer technique for in situ measurement of soil matric potential and root water extraction from a sandy soil

The suitability of microtensiometers to measure the spatial variation of soil matric potential and its diurnal change was tested in a pot experiment with pearl millet (Pennisetum americanum [L.] Leeke) in a sandy soil as the soil dried out.The temporal and spatial resolution of this technique allowed precise measurement of soil matric potential and thus estimation of soil water extraction from different compartments as well as from the whole rooting zone. The technique also allowed the measurement of rehydration of plants at night and root water uptake rate per unit soil volume or per unit root length. The precision of determination of root water uptake depended greatly on the accuracy of the estimate of hydraulic conductivity, which was derived from a bare soil and might be different for a cropped soil owing to aggregation induced by the root system. A linear relationship between root length and water uptake was found (r²=0.82), irrespective of variation in soil water content between compartments and despite the variation in root age, xylem differentiation and suberin formation expected to exist between different compartments of the rooting zone. As the experiment was carried out in a range of soil matric potentials between –4 and –30 kPa, drought stress did not occur. Further information at lower soil matric potentials are required, to address questions such as the importance of soil resistance for water uptake, or which portion of the root system has to be stressed to induce hormonal signals to the shoot. The microtensiometer technique can be applied to soil matric potentials up to –80 kPa.     

不同带长微喷带灌溉对土壤水分布与冬小麦耗水特性及产量的影响

于2010—2012年度冬小麦生长季,选用高产冬小麦品种济麦22,采用测墒补灌方式,设置40m(T40)、60m(T60)和80m(T80)3种带长的微喷带灌溉处理,研究不同带长微喷带灌溉对土壤水分分布及冬小麦耗水特性和产量的影响.结果表明:拔节期和开花期采用微喷带补灌,随微喷带带长缩短,灌溉水在土壤中的水平分布均匀系数显著增加.拔节期补灌,T40和T60处理在距畦首0～40m范围内各小麦行间的0～200cm土层土壤含水量均无显著差异;T80处理在距畦首38～40m、58～60m和78～80m处各小麦行间的0～200cm各土层土壤含水量变化规律一致,均表现为随距微喷带的距离增加而减小.T40处理的小麦在拔节至开花期间和开花至成熟期间分别对40～60cm和20～80cm土层土壤贮水的消耗量显著高于T60和T80处理,而对深层土壤贮水消耗量和总土壤贮水消耗量、开花期补灌水量、总灌水量和总耗水量显著低于T60和T80处理.随微喷带带长缩短,小麦籽粒产量、产量水分利用效率显著升高,而流量降低,在灌水量一定的情况下,单位时间内的有效灌溉面积减小.综合考虑小麦籽粒产量、水分利用效率和流量,40和60m是本试验条件下的适宜微喷带带长.     

Drip irrigation,soil characteristics and the root distribution and root activity of olive trees

A study was carried out on the root distribution and root activity of the olive tree (Olea Europaea, L., var. manzanillo) as influenced by drip irrigation and by several soil characteristics such as texture and depth. The experiments were conducted in two plots within a drip-irrigated grove of 20-year-old trees planted at 7×7 m spacing. One soil was a sandy loam, the other a clay-loam. Both cylinder and trench methods were used to determine root distribution. Labelling with ³²P was used to determine root activity. Under dryland conditions the adult tree adapted its rooting system, following the installation of a drip system, by concentrating the roots within the wet soil zones near the drippers. The highest root densities occur in those zones, down to a 0.6 m depth, the most abundant being the <0.5 mm diameter roots. The most intensive root activity was also found in that zone. For a given irrigation system, wet soil bulbs are more extensive and therefore root distribution expands to a larger soil volume when the soil is more clayey and with a hard calcareous pan present at about 0.8 m depth which prevents deep drainage.     

Distribution of resting spores of the Lymantria dispar pathogen Entomophaga maimaiga in soil and on bark

Cadavers of late instar Lymantria dispar (gypsy moth) larvae killed by the fungal pathogen Entomophaga maimaiga predominantly contain resting spores (azygospores). These cadavers frequently remain attached to tree trunks for several weeks before they detach and fall to the ground. Density gradient centrifugation was used to quantify resting spores in the soil and on tree bark. Titers of resting spores were extremely high at 0–10 cm from the base of the tree and the number decreased with distance from the trunk of the tree. Titers were also highest in the organic layer of the soil with numbers decreasing precipitously with increasing depth in the soil. While resting spores were obtained from tree bark, densities per unit area were much lower than those found in the organic soil layer at the base of the tree. Field bioassays were conducted with caged L. dispar larvae to compare infection levels with distance from the tree trunk as well as on the trunk. Highest infection levels were found at 50cm from the tree base with lowest infection on the tree trunk at 0.5 m height, although we expected the highest infection levels among larvae caged at the bases of trees, where highest spore titers occurred. Laboratory experiments demonstrated that L. dispar larvae exposed to resting spore- bearing soil at the soil surface became infected while larvae exposed to soil with resting spores buried at least 1 cm below the surface did not become infected.     

Significance and limits in the use of predawn leaf water potential for tree irrigation

Research in estimating the water status of crops is increasingly based on plant responses to water stress. Several indicators can now be used to estimate this response, the most widely available of which is leaf water potential (Ψ_LWP) as measured with a pressure chamber. For many annual crops, the predawn leaf water potential (Ψ_PLWP), assumed to represent the mean soil water potential next to the roots, is closely correlated to the relative transpiration rate, RT. A similar correlation also holds for young fruit trees grown in containers. However, exceptions to this rule are common when soil water content is markedly heterogeneous. Two experimental conditions were chosen to assess the validity of this correlation for heterogeneous soil water content: 1) young walnut trees in split-root containers. The heterogeneity was created by two unequal compartments (20% and 80% of total volume), of which only the smaller was irrigated and kept at a moisture content higher than field capacity (permanent drainage). 2) adult walnut trees in an orchard. In this case, soil water heterogeneity was achieved by limiting the amount of localised irrigation (20% of the irrigated control) which was applied every evening. Values of sap flux and of minimum and predawn leaf water potentials with homogeneous and heterogeneous soil water content were compared for trees grown in the orchard and in containers. In spite of intense drought reflected by very low RT or stem water potential, Ψ_PLWP of trees under heterogeneous moisture conditions remained high (between -0.2 and -0.4 MPa) both in the orchard and in containers. These values were higher than those usually considered critical under homogeneous soil conditions. A semi-quantitative model, based on the application of Ohm's analogy to split-root conditions, is proposed to explain the apparently conflicting results in the literature on the relation between Ψ_PLWP and soil water potential. This revised version was published online in August 2006 with corrections to the Cover Date.     

Response of total and nitrate-dissimilating bacteria to reduced N deposition in a spruce forest soil profile

A field-scale manipulation experiment conducted for 16 years in a Norway spruce forest at Solling, Central Germany, was used to follow the long-term response of total soil bacteria, nitrate reducers and denitrifiers under conditions of reduced N deposition. N was experimentally removed from throughfall by a roof construction ('clean rain plot'). We used substrate-induced respiration (SIR) to characterize the active fraction of soil microbial biomass and potential nitrate reduction to quantify the activity of nitrate reducers. The abundance of total bacteria, nitrate reducers and denitrifiers in different soil layers was analysed by quantitative PCR of 16S rRNA gene, nitrate reduction and denitrification genes. Reduced N deposition temporarily affected the active fraction of the total microbial community (SIR) as well as nitrate reductase activity. However, the size of the total, nitrate reducer and denitrifier communities did not respond to reduced N deposition. Soil depth and sampling date had a greater influence on the density and activity of soil microorganisms than reduced deposition. An increase in the nosZ /16S rRNA gene and nosZ/nirK ratios with soil depth suggests that the proportion of denitrifiers capable of reducing N₂O into N₂ is larger in the mineral soil layer than in the organic layer.     

荒漠绿洲边缘降水和地下水对白刺群落消长的影响

通过在甘肃民勤荒漠绿洲外围设固定样方连年观测,结合小区域地下水位动态监测和气象观测数据,定量研究了降水和地下水位变化对白刺群落消长的影响。结果表明,以地下水位逐年下降为主要驱动因素的绿洲外围的环境旱化,促使优势种白刺群落的发育适应环境,更具旱生性。在民勤绿洲外围,地下水位下降在近期内不是白刺生长的限制因素,在该区白刺将能较长时间生存,并发挥其优良的固沙、阻沙作用。民勤绿洲外围地区约１１０ｍｍ年降水     

Screening method for detection of hydrocarbon-oxidizing bacteria in oil-contaminated water and soil specimens

Toxic action of crude oil on the living world and ecosystems in general is a global problem of both aquatic and terrestrial environments. Bearing in mind the possibility of biodegradation of this toxicant, the procedures of determining counts and activity of cultivable microorganisms, and especially of bacteria responsible for degradation processes, are of great significance. The aim of this work was to study the possibility of modifying some solid media by adding triphenyltetrazolium chloride reagent as an indicator of the dehydrogenase activity, to develop a simple screening method for a simultaneous assessment of the count and activity of cultivable hydrocarbon-oxidizing bacteria in the oil-contaminated environments. The modified method appeared to be rapid and very suitable for the intended purposes.     

Use of repetitive DNA sequences to determine the persistence of enteropathogenic Escherichia coli in vegetables and in soil grown in fields treated with contaminated irrigation water

AIMS: Fresh fruits and vegetables are increasingly recognized as vectors for food-borne illness. On farm contamination through contaminated irrigation water is considered likely source of the pathogen for several outbreaks. The purpose of this study is to investigate the possible similarity of strains of Escherichia coli isolated from the soil and vegetables irrigated by treated wastewater. METHODS AND RESULTS: Seventy-five strains of enteropathogenic Escherichia coli isolated from vegetables, soil and irrigation water were tested for sensitivity to antibiotics and shown to be sensitive. The result of enterobacterial repetitive intergenic consensus (ERIC)-PCR shows similarities between analysed strains isolated from the three different samples. Moreover strains of E. coli isolated from vegetables over different periods of time have the same ERIC-PCR profile. CONCLUSIONS: The isolated strains of enteropathogenic E. coli can persist in soil and in vegetables growing in fields treated with contaminated irrigation water for an extended period of time. SIGNIFICANCE AND IMPACT OF THE STUDY: Contaminated irrigation water can transport pathogenic bacteria, which persists in the soil for a long period of time and contaminates the vegetables growing in the field irrigated by this contaminated water.     

Identifying the functional groups and the influence of synthetic chelators on Cd availability and microbial biomass carbon in Cd-contaminated soil

Synthetic chelators play an important role in boosting the microbial biomass carbon (MBC), dissolved organic carbon (DOC), and heavy metal solubility in a contaminated soil toward a sustainability of environment for agricultural crops. Castor plant was grown under different levels of Cd contaminated soil (?Cd and +Cd) following adding three chelating agents, ethylenediaminetetraacetic acid (H₄EDTA), nitriloacetic acid (H₃ NTA), and NH₄ citrate (ammonium citrate) to the soil at rates of 10, 15, and 25 mmol in 5 kg of soil per pot. The highest bioavailable Cd concentrations in soil and castor plant were obtained from NH₄ citrate and H₄EDTA treatments in the contaminated soil. Fourier transform infrared (FTIR) analysis showed that NH₄ citrate was the most effective chelator in Cd-contaminated soil. MBC and DOC contents were significantly increased and reached at 81.98–80.37 and 1.96–1.90 mg kg^?1 respectively, in the (H₃ NTA) and NH₄ citrate treatments in Cd-contaminated soil. Further research is needed to investigate the use of chelators in the phytoextraction of Cd-contaminated soils under field conditions and whether it may be beneficial in accelerating the phytoextraction of Cd through hyperaccumulating plants.