Comparison of the Temporal Variability of Enterococcal Clusters in Impacted Streams Using a Multiplex Polymerase Chain Reaction Procedure

Proceedings of the 2007 Georgia Water Resources Conference, March 27-29, 2007, Athens, Georgia.Understanding how fecal indicator bacteria and/or fecal indicator genotypes vary over time is important to determine the sources of fecal contamination. Enterococcus is one of the two indicators recommended by the EPA to monitor freshwaters for fecal contamination. Along with Escherichia coli, it has been used by a number of researchers to infer sources of fecal contamination, an area identified as microbial source tracking (MST). Our objective in this study was to identify changes in the seasonal distribution of enterococcal populations in streams directly impacted by cattle farming. The sites under study are located in Madison County, Ga., in farms where cows have unrestricted access to first order streams. Enterococci were counted and isolated monthly from water samples using membrane filtration. The isolates were identified using a multiplex PCR procedure. From a total of nine species identified in stream samples, only the most frequently observed species (E. faecalis, E. casseliflavus, E. flavescens, E. faecium and E. hirae) were used to develop groupings of enterococcal populations via cluster analysis. This analysis revealed that E. casseliflavus and E. faecalis dominated the enterococcal community during spring and fall, respectively. The cluster dominated by E. faecium seemed to increase during winter. This study indicates that enterococcal communities exhibit seasonal variability; and suggests that cluster analysis is a robust approach to identify this variability. In conclusion, to determine the true impact of certain farming operations on stream water quality using enterococcal species as indicators, it is important to consider the temporal variability of key enterococcal communities.Sponsored and Organized by: U.S. Geological Survey, Georgia Department of Natural Resources, Natural Resources Conservation Service, The University of Georgia, Georgia State University, Georgia Institute of TechnologyThis book was published by the Institute of Ecology, The University of Georgia, Athens, Georgia 30602-2202. The views and statements advanced in this publication are solely those of the authors and do not represent official views or policies of The University of Georgia, the U.S. Geological Survey, the Georgia Water Research Institute as authorized by the Water Resources Research Act of 1990 (P.L. 101-397) or the other conference sponsors

Fecal Coliform Bacteria TMDL Implementation on Cane Creek and Little Cane Creek in Oconee County, South Carolina

2008 S.C. Water Resources Conference - Addressing Water Challenges Facing the State and Regio

Conectividad de Java con bases de datos mediante invocación de objetos con métodos remotos (objetos RMI)

El desarrollo de aplicaciones distribuidas está teniendo cada vez más auge entre las empresas del mundo; esto se debe a la gran importancia que ha adquirido el Internet en los últimos años. Para dar solución a esta necesidad surgen arquitecturas distribuidas como RMI (Invocaci6n de Métodos Remotos). En este artículo se describe una de las principales arquitecturas empleadas para desarrollar aplicaciones basadas en objetos distribuidos, Java RMI. Además se presenta una comparación entre esta arquitectura y otras como CORBA, RPC y DCOM

Comunidades microbianas de solos de Cerrado sob vegetação nativa sujeita a queimadas prescritas e sob pastagem

O objetivo deste trabalho foi avaliar os efeitos de regimes de queima e cobertura vegetal sobre a dinâmica e a estrutura de comunidades microbianas do solo, por meio da análise de perfis de ácidos graxos de fosfolipídeos (PLFA). Compararam-se áreas nativas com diferentes coberturas vegetais (cerrado stricto sensu e campo sujo), sob diferentes regimes de queima e uma pastagem de capim-braquiária de 20 anos, no Planalto Central do Brasil. A biomassa microbiana foi maior nas parcelas com vegetação nativa que na pastagem, e os maiores valores mensais foram observados durante a estação chuvosa nas parcelas nativas. Apesar de não terem sido observadas diferenças significativas como consequência das queimadas ou entre as vegetações nativas, a análise de componentes principais separou as comunidades microbianas pela cobertura vegetal (nativa x pastagem) e pela sazonalidade (chuvosa x seca), tendo explicado 45,8 e 25,6%, respectivamente, da variabilidade de PLFA total. Mudanças na cobertura do solo e a sazonalidade na precipitação têm efeito significativo sobre a densidade total e a abundância de grupos de microrganismos do solo no Cerrado, especialmente bactérias Gram-negativas e Gram-positivasThe objective of this work was to evaluate the effects of fire regimes and vegetation cover on the structure and dynamics of soil microbial communities, through phospholipid fatty acid (PLFA) analysis. Comparisons were made between native areas with different woody covers ("cerrado stricto sensu" and "campo sujo"), under different fire regimes, and a 20-year-old active palisadegrass pasture in the Central Plateau of Brazil. Microbial biomass was higher in the native plots than in the pasture, and the highest monthly values were observed during the rainy season in the native plots. No significant differences were observed between fire regimes or between communities from the two native vegetation types. However, the principal component (PC) analysis separated the microbial communities by vegetation cover (native x pasture) and season (wet x dry), accounting for 45.8% (PC1 and PC3) and 25.6% (PC2 and PC3), respectively, of the total PLFA variability. Changes in land cover and seasonal rainfall in Cerrado ecosystems have significant effects on the total density of soil microorganisms and on the abundance of microbial groups, especially Gram-negative and Gram-positive bacteria

Stoichiometric linkages between plant litter, trophic interactions and nitrogen mineralization across the litter-soil interface

The common notion for describing N mineralization in models is that it results from decomposer organisms trying to meet their stoichiometric demands based on their own elemental composition and that of the resource. However, in addition to influencing C and nutrient availability, plant litter also influences the composition of both the litter and mineral soil community – importantly not in the same manner – resulting in altered trophic interactions. Since decomposer groups and their consumers vary in their elemental composition and demands, a change in composition and abundance of soil functional groups may result in a change in the stoichiometry of the whole soil food web, thus altering their stoichiometric relations with the available resource with potential functional consequences. We use experimental data and quantitative food web modeling to investigate the impact of the changes in the litter and soil food webs brought about by the differing stoichiometry of plant litter on (a) N mineralization, (b) the contribution of different functional groups to mineralization, and (c) the stoichiometric flexibility of the system, assessed as the ability to mineralize materials with different stoichiometry. Our simulations suggested that the effects of litter stoichiometry on trophic interactions, their impacts on N mineralization and the relative contribution of functional groups may not behave as a continuum across the litter and soil interface. Further, changes in food webs associated with variation in plant stoichiometric traits can influence the relative importance of functional groups, which given their particular stoichiometric demands may affect ecosystem-level N cycling. Our results suggested that litter materials of intermediate N contents, or litter mixtures encompassing materials with different nutrient contents and thus resulting in mixtures of intermediate stoichiometry, may promote food webs that are better suited to deal with changing substrate stoichiometry

The influence of litter composition across the litter-soil interface on mass loss, nitrogen dynamics and the decomposer community

Many studies have investigated the influence of plant litter species composition on decomposition, but results have been context-dependent. Litter and soil are considered to constitute a decomposition continuum, but whether litter and soil ecosystems respond to litter identity and mixing in the same manner is unsure. In a field experiment utilizing 5 litter species and their mixture, we investigated whether the effects of litter identity and mixing on mass loss, nutrient dynamics, and decomposer communities are consistent across the litter-soil interface. In monoculture, mass loss and nitrogen (N) dynamics in the litter layer corresponded to the underlying soil N availability, demonstrating the continuum of resources from litter to soil. Litter microbial biomass and mesofauna abundance tended to be greater on litter with a faster decay rate and greater N release. However, soil decomposer abundance and diversity were not greater with higher soil N, causing litter and soil communities to respond differently to litter identity. Non-additive mass loss and N dynamics were observed after 6 months, and were correlated with non-additive litter microbial community composition and litter mesofauna communities, but all other aspects of the litter community and all measures of the soil community were additive. Decomposer communities and N dynamics did not respond similarly to the litter mixture across the litter-soil interface. This study is one of the few to comprehensively examine how a litter mixture influences decomposition dynamics and communities across the soil-litter interface, including multiple taxa and trophic levels. Our results demonstrate that processes associated with decomposition are decoupled for litter and soil, particularly in that litter showed non-additivity in mass loss, N release and decomposer community, but soil responses were largely additive

Sources and Drivers of ARGs in Urban Streams in Atlanta, Georgia, USA

The spread of antibiotic resistance genes (ARGs) in the aquatic environment is an emerging concern in the interest of protecting public health. Stemming the environmental dissemination of ARGs will require a better understanding of the sources and drivers of ARGs in the water environment. In this study, we used direct measurement of sewage-associated molecular markers, the class 1 integron gene, standard water quality parameters, and watershed characteristics to evaluate the sources and drivers of ARGs in an urban watershed impacted by a gradient of human activities. Quantitative polymerase chain reaction (qPCR) was used to quantify the abundance of the sewage-associated HF183, the E. coli fecal indicator, class 1 integron gene (int1), and the ARGs sulI, sulII, tetW, tetM, ampC, and blaSHV in stream water samples collected from the Proctor Creek watershed in Atlanta, Georgia. Our findings show that ARGs were widely distributed, with detection frequencies of 96% (sulI and sulII), 82% (tetW and tetM), and 49% (ampC and blaSHV). All the ARGs were positively and significantly correlated (r > 0.5) with the HF183 and E. coli markers. Non-linear machine learning models developed using generalized boosting show that more than 70% of the variation in ARG loads in the watershed could be explained by fecal source loading, with other factors such as class 1 integron, which is associated with acquired antibiotic resistance, and environmental factors contributing < 30% to ARG variation. These results suggest that input from fecal sources is a more critical driver of ARG dissemination than environmental stressors or horizontal gene transfer in aquatic environments highly impacted by anthropogenic pollution. Finally, our results provide local watershed managers and stakeholders with information to mitigate the burden of ARGs and fecal bacteria in urban streams

Soil fauna alter the effects of litter composition on nitrogen cycling in a mineral soil

Plant chemical composition and the soil community are known to influence litter and soil organic matter decomposition. Although these two factors are likely to interact, their mechanisms and outcomes of interaction are not well understood. Studies of their interactive effects are rare and usually focus on carbon dynamics of litter, while nutrient dynamics in the underlying soil have been ignored. A potential mechanism of interaction stems from the role fauna plays in regulating availability of litter-derived materials in the mineral soil. We investigated the role of soil fauna (meso, macro) in determining the effect of surface-litter chemical composition on nitrogen mineralization and on the micro-food web in mineral soils. In a field setting we exposed mineral soil to six types of surface-applied litter spanning wide ranges of multiple quality parameters and restricted the access of larger soil animals to the soils underlying these litters. Over six months we assessed litter mass and nitrogen loss, nitrogen mineralization rates in the mineral soils, and soil microbes and microfauna. We found evidence that the structure of the soil community can alter the effect of surface-litter chemical composition on nitrogen dynamics in the mineral soil. In particular, we found that the presence of members of the meso- and macrofauna can magnify the control of nitrogen mineralization by litter quality and that this effect is time dependent. While fauna were able to affect the size of the micro-food web they did not impact the effect of litter composition on the abundance of the members of the micro-food web. By enhancing the strength of the impact of litter quality on nitrogen dynamics, the larger fauna can alter nitrogen availability and its temporal dynamics which, in turn, can have important implications for ecosystem productivity. These findings contribute to evidence demonstrating that soil fauna shape plant litter effects on ecosystem function

Conectividad de Java con bases de datos mediante invocación de objetos con métodos remotos (objetos RMI)

El desarrollo de aplicaciones distribuidas está teniendo cada vez más auge entre las empresas del mundo; esto se debe a la gran importancia que ha adquirido el Internet en los últimos años. Para dar solución a esta necesidad surgen arquitecturas distribuidas como RMI (Invocaci6n de Métodos Remotos). En este artículo se describe una de las principales arquitecturas empleadas para desarrollar aplicaciones basadas en objetos distribuidos, Java RMI. Además se presenta una comparación entre esta arquitectura y otras como CORBA, RPC y DCOM

Transcriptome Changes of Escherichia coli, Enterococcus faecalis, and Escherichia coli O157:H7 Laboratory Strains in Response to Photo-Degraded DOM

In this study, we investigated gene expression changes in three bacterial strains (Escherichia coli C3000, Escherichia coli O157:H7 B6914, and Enterococcus faecalis ATCC 29212), commonly used as indicators of water quality and as control strains in clinical, food, and water microbiology laboratories. Bacterial transcriptome responses from pure cultures were monitored in microcosms containing water amended with manure-derived dissolved organic matter (DOM), previously exposed to simulated sunlight for 12 h. We used RNA sequencing (RNA-seq) and quantitative real-time reverse transcriptase (qRT-PCR) to compare differentially expressed temporal transcripts between bacteria incubated in microcosms containing sunlight irradiated and non-irradiated DOM, for up to 24 h. In addition, we used whole genome sequencing simultaneously with RNA-seq to identify single nucleotide variants (SNV) acquired in bacterial populations during incubation. These results indicate that E. coli and E. faecalis have different mechanisms for removal of reactive oxygen species (ROS) produced from irradiated DOM. They are also able to produce micromolar concentrations of H2O2 from non-irradiated DOM, that should be detrimental to other bacteria present in the environment. Notably, this study provides an assessment of the role of two conjugative plasmids carried by the E. faecalis and highlights the differences in the overall survival dynamics of environmentally-relevant bacteria in the presence of naturally-produced ROS