A threshold area ratio of organic to conventional agriculture causes recurrent pathogen outbreaks in organic agriculture

Conventional agriculture uses herbicides, pesticides, and chemical fertilizers that have the potential to pollute the surrounding land, air and water. Organic agriculture tries to avoid using these and promotes an environmentally friendly approach to agriculture. Instead of relying on herbicides, pesticides and chemical fertilizers, organic agriculture promotes a whole system approach to managing weeds, pests and nutrients, while regulating permitted amendments. In this paper, we consider the effect of increasing the total area of agricultural land under organic practices, against a background of conventional agriculture. We hypothesized that at a regional scale, organic agriculture plots benefit from existing in a background of conventional agriculture, that maintains low levels of pathogens through pesticide applications. We model pathogen dispersal with a diffusive logistic equation in which the growth/death rate is spatially heterogeneous. We find that if the ratio of the organic plots to conventional plots remains below a certain threshold l_c, the pest population is kept small. Above this threshold, the pest population in the organic plots grows rapidly. In this case, the area in organic agriculture will act as a source of pest to the surrounding region, and will always infect organic plots as they become more closely spaced. Repeated localized epidemics of pest outbreaks threaten global food security by reducing crop yields and increasing price volatility. We recommend that regional estimates of this threshold are necessary to manage the growth of organic agriculture region by region.     

Macromolecule mediated transport and retention of Escherichia coli O157:H7 in saturated porous media

The role of extracellular macromolecules on Escherichia coli O157:H7 transport and retention was investigated in saturated porous media. To compare the relative transport and retention of E. coli cells that are macromolecule rich and deficient, macromolecules were partially cleaved using a proteolytic enzyme. Characterization of bacterial cell surfaces, cell aggregation, and experiments in a packed sand column were conducted over a range of ionic strength (IS). The results showed that macromolecule-related interactions contribute to retention of E. coli O157:H7 and are strongly linked to solution IS. Under low IS conditions (IS ≤ 0.1 mM), partial removal of the macromolecules resulted in a more negative electrophoretic mobility of cells and created more unfavorable conditions for cell-quartz and cell-cell interactions as suggested by Derjaguin-Landau-Verwey-Overbeek (DLVO) interaction energy profiles and cell aggregation kinetics. Consequently, less retention was observed for enzyme treated cells in the corresponding column experiments. In addition, a time-dependent deposition process (i.e., ripening) was observed for untreated cells, but not for treated cells, supporting the fact that the macromolecules enhanced cell-cell interactions. Additional column experiments for untreated cells under favorable conditions (IS ≥ 1 mM) showed that a significant amount of the cells were reversibly retained in the column, which contradicts predictions of DLVO theory. Furthermore, a non-monotonic cell retention profile was observed under favorable attachment conditions. These observations indicated that the presence of macromolecules hindered irreversible interactions between the cells and the quartz surface.     

Pyrosequencing of the 16S rRNA gene to reveal bacterial pathogen diversity in biosolids

Given the potential for a variety of bacterial pathogens to occur in variably stabilized sewage sludge (biosolids), an understanding of pathogen diversity and abundance is necessary for accurate assessment of infective risk when these products are land applied. 16S rDNA was PCR amplified from genomic DNA extracted from municipal wastewater residuals (mesophilic- and thermophilic-phased anaerobic digestion (MAD and TPAD), composting (COM)), and agricultural soil (SOIL), and these amplicons were sequenced using massively parallel pyrosequencing technology. Resulting libraries contained an average of 30,893 16S rDNA sequences per sample with an average length of 392 bases. FASTUNIFRAC-based comparisons of population phylogenetic distance demonstrated similarities between the populations of different treatment plants performing the same stabilization method (e.g. different MAD samples), and population differences among samples from different biosolids stabilization methods (COM, MAD, and TPAD). Based on a 0.03 Jukes-Cantor distance to 80 potential bacterial pathogens, all samples contained pathogens and enrichment ranged from 0.02% to 0.1% of sequences. Most (61%) species identified were opportunistic pathogens of the genera Clostridium and Mycobacterium. As risk sciences continue to evolve to address scenarios that include multiple pathogen exposure, the analysis described here can be used to determine the diversity of pathogens in an environmental sample. This work provides guidance for prioritizing subsequent culturable and quantitative analysis, and for the first time, ensuring that potentially significant pathogens are not left out of risk estimations.     

Optimization of PCR-based methods for rapid detection of Campylobacter jejuni, Campylobacter coli and Yersinia enterocolitica serovar 0:3 in wastewater samples

PCR-based methods were evaluated for their adequacy to assess the removal of pathogens from wastewater samples. For the development and optimization of the methods, samples were taken at two different sites from two different constructed wetlands. Campylobacter jejuni/coli and Yersinia enterocolitica serogroup 0:3 were selected as model pathogens and Enterococcus faecalis as a standard microbiological indicator. The chosen PCR protocols were optimized for wastewater DNA extracts in order to obtain high sensitivity and reproducibility independently of the background flora. All PCR protocols were successfully performed and reproducible with a background of up to 10(10) nontarget cells per reaction. Five cells of Y. enterocolitica, 50 cells of C. jejuni/coli, and 500 cells of E. faecalis per 100ml treated water could be detected. The method detection limit in the settled wastewater was higher: 200 cells per 100ml for Y. enterocolitica, 2000 cells per 100ml for C. jejuni/coli, and 20,000 cells per 100ml for E. faecalis. C. jejuni/coli and Y. enterocolitica PCRs were adapted to municipal wastewater, with higher loads of potential PCR inhibitors. Sensitivity was lower for this type of wastewater: 200 cells of Y. enterocolitica and 2000 cells of C. jejuni/coli were detected per 100ml treated wastewater, 2500 cells of Y. enterocolitica and 25,000 cells of C. jejuni/coli per 100ml settled wastewater. The developed PCR methods enable the detection of C. jejuni/coli, Y. enterocolitica serogroup 0:3 and E. faecalis within 12h. They show specificity, reproducibility and low detection limits for the investigated pathogens.     

瓜类枯萎病菌生物学特性与药剂测定研究

对瓜类枯萎病的3种专化型即黄瓜专化型（F.oxysporum f.sp.cucumerinum),西瓜专化型（F.oxysporum f.sp.niveum)和甜瓜专化型（F.oxysporm f.sp.melonis)进行了形态特点,生物学特性与药剂测定等室内比较研究。结果表明,枯萎病菌的3种专化型,在形态特征,培养性状以及对杀菌剂的敏感性等方面,既有明显的相似点,也有不同之处,菌丝生长的温度范围均为5-35℃,以25℃为最适;在20-30℃的温度范围内,孢子萌发率较高,病菌3种专化型的产孢量与温度的相关性不明显,pH值为6最适于孢子萌发,健根保,苗菌敌,无敌,根必治和重茬保5种药剂,对瓜类枯萎病菌菌丝生长的抑制效果较好。     

不同来源的副溶血性弧菌定性定量分析及毒素基因检测

目的研究受污染海产品、海产品养殖水及食物中毒临床分离的副溶血性弧菌的生化特性、种特异性基因片段检测方法的应用和毒素基因携带情况。方法66株副溶血性弧菌采用全自动微生物鉴定、MPN值定量、氯化物含量(离子色谱法)、R72H基因片段和tdh、trh毒素基因PCR检测等方法进行研究。结果定性定量分析方面,98.5%的菌株不发酵阿拉伯糖;牡蛎样本MPN值均为≥24000/100g,并检出2株携带trh毒素基因;23/30份养殖水检出副溶血性弧菌,其中海丰监测点6份阳性水样的氯化物含量均值仅为3160mg/L,显著低于实验室培养的30g/L的含量。基因检测方面,VITEK鉴定R72H基因片段结果与检测相符;13株食物中毒临床分离株均携带tdh毒素基因;6.7%的海产品分离株、15.4%食物中毒临床来源株同时携带两种毒素基因。结论通过从海产品和养殖水的检出情况以及分离株的生化特性、毒素基因资料,为副溶血性弧菌食物中毒溯源提供重要的数据源。     

Opportunities for mitigating pathogen contamination during on-farm food production

Fruits, vegetables, and meat are susceptible to contamination by foodborne pathogens at many points from production through preparation in the home. This review will largely highlight approaches and progress made in the last five years to address strategies to reduce pathogen contamination in animal production but will also touch on the emerging field of preharvest produce food safety. Mitigation strategies can be divided into those that address pathogen reduction in the environment and those that target reduction/elimination of pathogen contamination in animals or plants. The former strategy has been encompassed in studies evaluating sanitation treatments of facilities as well as in numerous epidemiologic risk assessment studies (both on-farm assessments and computer simulation models) that identify management practices that impact pathogen prevalence in animals. Interventions to significantly reduce pathogen exposure via feed or water are dependent on their role as a significant contributor to pathogen contamination in the animal production system. In addition, inconsistent results obtained with interventions of dietary additives or formulation modifications (grain versus forage; inclusion of distiller's grains) on pathogen prevalence in animals have been attributed to a range of factors including target organism, grain type, level of inclusion, the animal's health or stress level, and ability to survive the gastric acidic conditions. Recent attempts to microencapsulate organic acids or bacteriophage within feed have met with only marginal improvements in reducing pathogen carriage in animals but this approach may have greater potential with other antimicrobial additives (i.e., essential oils). Bacteriophage therapy, in general, can significantly reduce pathogen carriage in animals but based on its transient nature and the potential for development of phage-resistant subpopulations, this approach should be administered to animals just prior to slaughter and preferably to animals that are suspected “super-shedders”. Other promising on-farm intervention approaches have included breeding for pathogen resistance, vaccines, and dietary bacteriocins. To optimize interventions on a cost basis, studies have also determined that application of dietary interventions at specific time points in the animal's production cycle is a useful strategy to reduce pathogen carriage (e.g., probiotics to fertilized eggs and acidified feed to fattening swine). In conclusion, applicable management and intervention strategies may vary depending on the type of food under production; however, it is important to consider from a holistic view how any new intervention strategies will affect the overall production system in order to maintain a successful, efficient food production environment.     

Mechanisms of microwave irradiation involved in the destruction of fecal coliforms from biosolids

Microwaves have been found to be effective in destructing pathogens in sewage sludge (biosolids) (75th Annual Water Environment Federation Conference, Chicago, September 29-October 2, 2002; Third World Water Congress, International Water Association, Melbourne, Australia, April 7-12, 2002). Mechanisms and roles of microwaves on fecal coliform destruction were investigated using bacterial viability tests, electron transport system (ETS) and beta-galactosidase activity assays, gel electrophoresis, and genomic deoxyribonucleic acid (DNA) optical density (OD) measurements with fecal coliforms isolated from biosolids. Bacterial viability tests demonstrated cell membrane damage as microwave irradiation intensity and temperature increased. Above 60+/-3 degrees C, viable cells were rarely found when pure fecal coliforms were irradiated with microwaves. ETS and beta-galactosidase activity assays revealed increased activity for externally heated samples due to fecal coliform growth but decreased activity for microwave-irradiated samples as temperature was increased from 20 degrees C to 57 degrees C, indicating other destruction mechanisms besides heating. Between 57 degrees C and 68 degrees C, microwave irradiation led to a more rapid decrease in activity than external heating by convection. Above 68 degrees C, bacterial activity almost ceased for both pretreatments. DNA bands in gel electrophoresis tests and OD of genomic DNA decreased more rapidly for microwave-irradiated samples than for externally heated samples, implying that microwaves disrupted DNA in fecal coliform cells at lower temperatures than external heating. Microwave irradiation of sludge appears to be a viable and economical method of destructing pathogens and generating environmentally safe sludge.