Watershed monitoring and the TMDL modeling to assess bacterial loading in estuarine environments to improve management

Shellfish bed closures along the North Carolina coast have increased over the years seemingly concurrent with increases in population (Mallin 2000). More and faster flowing storm water has come to mean more bacteria, and fecal indicator bacterial (FIB) standards for shellfish harvesting are often exceeded when no source of contamination is readily apparent (Kator and Rhodes, 1994). Could management reduce bacterial loads if the source of the bacteria where known? Several potentially useful methods for differentiating human versus animal pollution sources have emerged including Ribotyping and Multiple Antibiotic Resistance (MAR) (US EPA, 2005). Total Maximum Daily Load (TMDL) studies on bacterial sources have been conducted for streams in NC mountain and Piedmont areas (U.S. EPA, 1991 and 2005) and are likely to be mandated for coastal waters. TMDL analysis estimates allowable pollutant loads and allocates them to known sources so management actions may be taken to restore water to its intended uses (U.S. EPA, 1991 and 2005). This project sought first to quantify and compare fecal contamination levels for three different types of land use on the coast, and second, to apply MAR and ribotyping techniques and assess their effectiveness for indentifying bacterial sources. Third, results from these studies would be applied to one watershed to develop a case study coastal TMDL. All three watershed study areas are within Carteret County, North Carolina. Jumping Run Creek and Pettiford Creek are within the White Oak River Basin management unit whereas the South River falls within the Neuse River Basin. Jumping Run Creek watershed encompasses approximately 320 ha. Its watershed was a dense, coastal pocosin on sandy, relic dune ridges, but current land uses are primarily medium density residential. Pettiford Creek is in the Croatan National Forest, is 1133 ha. and is basically undeveloped. The third study area is on Open Grounds Farm in the South River watershed. Half of the 630 ha. watershed is under cultivation with most under active water control (flashboard risers). The remaining portion is forested silviculture.(PDF contains 4 pages)

Using microbiological tracers to assess the impact of winter land use restrictions on the quality of stream headwaters in a small catchment.

Diverse land use activities can elevate risk of microbiological contamination entering stream headwaters. Spatially distributed water quality monitoring carried out across a 17km(2) agricultural catchment aimed to characterize microbiological contamination reaching surface water and investigate whether winter agricultural land use restrictions proved effective in addressing water quality degradation. Combined flow and concentration data revealed no significant difference in fecal indicator organism (FIO) fluxes in base flow samples collected during the open and prohibited periods for spreading organic fertilizer, while relative concentrations of Escherichia coli, fecal streptococci and sulfite reducing bacteria indicated consistently fresh fecal pollution reached aquatic receptors during both periods. Microbial source tracking, employing Bacteroides 16S rRNA gene markers, demonstrated a dominance of bovine fecal waste in river water samples upstream of a wastewater treatment plant discharge during open periods. This contrasted with responses during prohibited periods where human-derived signatures dominated. Differences in microbiological signature, when viewed with hydrological data, suggested that increasing groundwater levels restricted vertical infiltration of effluent from on-site wastewater treatment systems and diverted it to drains and surface water. Study results reflect seasonality of contaminant inputs, while suggesting winter land use restrictions can be effective in limiting impacts of agricultural wastes to base flow water quality.

Bactérias coliformes totais e coliformes de origem fecal em águas usadas na dessedentação de animais

Foram colhidas 105 amostras de água sendo 44 de mananciais e de 61 bebedouros, a partir das quais foram realizadas as determinações dos NMP (Número Mais Provável) de bactérias coliformes totais e fecais. de acordo com a Portaria GM/0013 de 15 de janeiro de 1976, da Secretaria Especial do Meio Ambiente do Ministério do Interior, os 44 mananciais revelaram-se dentro dos parâmetros estabelecidos. Embora não haja referência com relação a bebedouros, aplicando-se os valores estabelecidos para mananciais, das 61 amostras estudadas, somente 6 (9,8%) não poderiam ser usadas para a dessedentação de animais. Das 105 amostras analisadas, verificou-se que as condições sanitárias revelaram-se não satisfatórias em 6 (5,7%) amostras de bebedouros quanto a coliformes fecais, sendo 3 (2,8%) também com relação a coliformes totais.

Assessment of anaerobic sewage sludge quality for agricultural application after metal bioleaching

The effects of metal bioleaching on nutrient solubilization, especially nitrogen and phosphorous, from anaerobically-digested sewage sludge were investigated in this work. The assessment of the sanitary quality of the anaerobic sludge after bioleaching was also carried out by enumerating indicator (total coliforms, fecal coliforms, and fecal streptococci) and total heterotrophic bacteria. The experiments of bioleaching were performed using indigenous sulphur-oxidizing bacteria (Thiobacillus spp.) as inoculum and samples of anaerobically-digested sludge. Nitrogen and phosphorous solubilization from sewage sludge was assessed by measuring, respectively, the concentration of Total Kjeldahl Nitrogen, ammonia, nitrate/nitrite, and soluble and total phosphorous before and after the bioleaching assays. At the end of the experiment, after 4 days of incubation (final pH of 1.4), the following metal solubilization yields were obtained: zinc, 91%; nickel, 87%; copper, 79%; lead, 52%; and chromium, 42%. As a result of sludge acidification, the viable counts of selected indicator bacteria were decreased to below the detection limit (4 × 103 cfu 100 ml-1), followed by an increase in the mineral fraction of nitrogen (from 6 to 10%) and in the soluble fraction of phosphorous (from 15 to 30%). Although some loss of sludge nutrients can occur during solid-liquid separation following bioleaching, its beneficial effects as metal removal and reduction of pathogenic bacteria are sufficient to consider the potential of this treatment before sludge disposal onto agricultural fields.

Detecção de adenovírus humanos em amostras de água superficial e esgoto não tratado oriundas de diversos ecossistemas aquáticos da cidade de Belém-PA

Os vírus entéricos são importantes agentes de doenças de veiculação hídrica. Entre esses, os adenovirus humanos (HAdV) assumiram importância por serem um dos principais causadores de gastrenterite em crianças menores de cinco anos e pela sua maior resistência a fatores físicos e químicos em detrimento a outros vírus no ambiente. Várias pesquisas têm demonstrado ausência de relação entre a presença de bactérias indicadoras e vírus. Diante disso, diversos autores têm sugerido a inclusão desses agentes como potenciais indicadores de contaminação viral e fecal da água. O objetivo desse trabalho foi detectar a presença de HAdV em amostras de água e esgoto não tratado oriundas de diversos ecossistemas aquáticos da cidade de Belém-PA. Foram selecionados seis pontos de amostragem, dentre eles um esgoto não tratado: Esgoto do UNA e cinco coleções hídricas: Porto do Açaí, Ver-o-Peso, Igarapé Tucunduba, Lago Bolonha e Lago Água Preta. Foi feita uma coleta mensal de dois litros de água em cada ponto durante 24 meses consecutivos, de nov/2008 a out/2010, totalizando 144 amostras. Foi utilizada água destilada autoclavada para controle negativo de cada ponto em todos os testes utilizados. As amostras foram concentradas pelo método de adsorção-eluição e posteriormente centrifugadas para a obtenção de dois mL. O DNA foi extraído pelo kit comercial Qiagen. Para a detecção molecular foram empregadas a Reação em Cadeia Mediada pela Polimerase (PCR convencional) e a PCR em tempo real, sendo utilizados iniciadores e sondas específicos que amplificam um gene do hexon de 301 e 96 pb, respectivamente. Visando-se melhorar o produto amplificado para sequenciamento genômico, algumas amostras positivas pela PCR convencional foram submetidas à Nested-PCR com a utilização de mais um par de iniciadores que amplificam uma região interna de 171 pb. Amostras de água e esgoto foram sequenciadas, analisadas e comparadas a outras obtidas no GeneBank. Os HAdV foram detectados em 59% (85/144) das amostras de água superficial e esgoto não tratado, sendo que a positividade obtida pela PCR convencional foi de 22,9% (33/144) e pela PCR em tempo real de 58,7% (84/143). A primeira detectou o agente apenas nas amostras do igarapé Tucunduba (62,5%) e do esgoto do UNA (75%) e a segunda em amostras provenientes dos seis pontos de coleta (com uma variação de 25 a 100%). O agente foi detectado em todos os 24 meses do estudo, estando presentes em pelo menos dois pontos, mensalmente. A PCR em tempo real se mostrou mais sensível nesse estudo, tendo encontrado o agente em 36,4% (52/143) das amostras não detectadas pela PCR convencional. Das oito amostras genotipadas todas pertencem à espécie F, sendo quatro referentes ao sorotipo 40 e quatro ao 41. Nossos resultados confirmam a alta circulação desse patógeno nas águas superficiais e esgoto da cidade, sugerindo a inclusão dos HAdV como bons indicadores de contaminação viral e fecal da água. A pesquisa desses vírus em ambientes aquáticos é pioneira em Belém e tais resultados são de relevante importância para as políticas de saúde pública e ambiental, servindo como base para estudos complementares nessa área.

Inactivation and structural changes of E. Cloacae and B. Subtilis Endospores during IR laser water treatment

IR radiation has been studied for micro-organism inactivation of bacterial spores on metal substrates [1] and on metal and paper substrates [2]. A near-point near infrared laser water treatment apparatus for use in dental hand-pieces was also developed [3]. To date water sterilisation research using a mid-IR laser technique is very rare. According to the World Health Organisation [4], examinations for faecal indicator bacteria remain the most sensitive and specific way of assessing the hygienic quality of water. Bacteria that fall into this group are E. coli, other coliform bacteria (including E. cloacae) and to a lesser extent, faecal streptococci [5]. Protozoan cysts from organisms which cause giardiasis are the most frequently identified cause of waterborne diseases in developed countries [6,7]. The use of aerobic bacterial endospores to monitor the efficiency of various water treatments has been shown to provide a reliable and simple indicator of overall performance of water treatment[8,9].The efficacy of IR radiation for water disinfection compared to UV treatment has been further investigated in the present study. In addition FTIR spectroscopy in conjunction with Principle Component Analysis was used to characterise structural changes within the bacterial cells and endospores following IR laser treatment. Changes in carbohydrate content of E. cloacae following IR laser treatment were observed.

Distribution and taxonomy of Enterococci from the Davis Station wastewater discharge, Antarctica

This project assessed the potential impact of untreated sewage release in a near-shore marine environment of Antarctica through the distribution and characterisation of the faecal indicator bacteria Enterococcus. Antibiotic resistance and genome sequencing analyses revealed that enterococci resistant to multiple antibiotics closely related to clinical pathogens were introduced to the pristine Antarctic environment by Australia's Davis station.

Support for Integrated Ecosystem Assessments of NOAA’s National Estuarine Research Reserves System (NERRS), Volume I: The Impacts of Coastal Development on the Ecology and Human Well-being of Tidal Creek Ecosystems of the US Southeast

A study was conducted, in association with the Sapelo Island and North Carolina National Estuarine Research Reserves (NERRs), to evaluate the impacts of coastal development on sentinel habitats (e.g., tidal creek ecosystems), including potential impacts to human health and well-being. Uplands associated with southeastern tidal creeks and the salt marshes they drain are popular locations for building homes, resorts, and recreational facilities because of the high quality of life and mild climate associated with these environments. Tidal creeks form part of the estuarine ecosystem characterized by high biological productivity, great ecological value, complex environmental gradients, and numerous interconnected processes. This research combined a watershed-level study integrating ecological, public health and human dimension attributes with watershed-level land use data. The approach used for this research was based upon a comparative watershed and ecosystem approach that sampled tidal creek networks draining developed watersheds (e.g., suburban, urban, and industrial) as well as undeveloped sites. The primary objective of this work was to clearly define the relationships between coastal development with its concomitant land use changes and non-point source pollution loading and the ecological and human health and well-being status of tidal creek ecosystems. Nineteen tidal creek systems, located along the southeastern United States coast from southern North Carolina to southern Georgia, were sampled during summer (June-August), 2005 and 2006. Within each system, creeks were divided into two primary segments based upon tidal zoning: intertidal (i.e., shallow, narrow headwater sections) and subtidal (i.e., deeper and wider sections), and watersheds were delineated for each segment. In total, we report findings on 24 intertidal and 19 subtidal creeks. Indicators sampled throughout each creek included water quality (e.g., dissolved oxygen concentration, salinity, nutrients, chlorophyll-a levels), sediment quality (e.g., characteristics, contaminants levels including emerging contaminants), pathogen and viral indicators, and abundance and genetic responses of biological resources (e.g., macrobenthic and nektonic communities, shellfish tissue contaminants, oyster microarray responses). For many indicators, the intertidally-dominated or headwater portions of tidal creeks were found to respond differently than the subtidally-dominated or larger and deeper portions of tidal creeks. Study results indicate that the integrity and productivity of headwater tidal creeks were impaired by land use changes and associated non-point source pollution, suggesting these habitats are valuable early warning sentinels of ensuing ecological impacts and potential public health threats. For these headwater creeks, this research has assisted the validation of a previously developed conceptual model for the southeastern US region. This conceptual model identified adverse changes that generally occurred in the physical and chemical environment (e.g., water quality indicators such as indicator bacteria for sewage pollution or sediment chemical contamination) when impervious cover levels in the watershed reach 10-20%. Ecological characteristics responded and were generally impaired when impervious cover levels exceed 20-30%. Estimates of impervious cover levels defining where human uses are impaired are currently being determined, but it appears that shellfish bed closures and the flooding vulnerability of headwater regions become a concern when impervious cover values exceed 10-30%. This information can be used to forecast the impacts of changing land use patterns on tidal creek environmental quality as well as associated human health and well-being. In addition, this study applied tools and technologies that are adaptable, transferable, and repeatable among the high quality NERRS sites as comparable reference entities to other nearby developed coastal watersheds. The findings herein will be of value in addressing local, regional and national needs for understanding multiple stressor (anthropogenic and human impacts) effects upon estuarine ecosystems and response trends in ecosystem condition with changing coastal impacts (i.e., development, climate change). (PDF contaions 88 pages)

Fibroblastos gengivais humanos em co-cultura com Aggregatibacter actinomycetemcomitans lisogênico induzem a liberação de fago

A relação entre bacteriófagos e virulência bacteriana é um modelo muito intrigante e pouco estudado na patogênese periodontal, uma vez que um patógeno periodontal pode ser lisogênico. O objetivo do nosso estudo é determinar a capacidade de fibroblastos gengivais humanos de induzir as cepas lisogênicas de Aggregatibacter actinomycetemcomitans. Dois experimentos foram realizados seguidos da titulação de fago. Os experimentos consistiram da co-cultura com fibroblastos gengivais humanos e três cepas de Aa [Aa29524, Aa2112, Aa29524(Ø2112)], não lisogênica, lisogênica e lisogênica induzida em laboratório, respectivamente. Em três momentos distintos (no experimento 1: 0, 2 e 4 horas; e no experimento 2: 2, 4 e 6 horas), o sobrenadante da co-cultura foi filtrado e cultivado overnight com a bactéria indicadora (Aa29524) e analisado para a capacidade de lisar a célula indicadora. Em ambos os experimentos, o sobrenadante da co-cultura de fibroblastos gengivais humanos com Aa lisogênico e Aa lisogênico induzido em laboratório, ao ser cultivado com a bactéria indicadora, promoveu lise da mesma, resultando no aumento da produção de fago. Pode-se concluir que, nesse estudo os fibroblastos gengivais humanos foram capazes de induzir cepas lisogênicas de Aggregatibacter actinomycetemcomitans.

Support for integrated ecosystem assessments of NOAA’s National Estuarine Research Reserve System (NERRS): assessment of ecological condition and stressor impacts in subtidal waters of the Sapelo Island National Estuarine Research Reserve

A study was conducted in June 2009 to assess the current status of ecological condition and potential human-health risks throughout subtidal estuarine waters of the Sapelo Island National Estuarine Research Reserve (SINERR) along the coast of Georgia. Samples were collected for multiple indicators of ecosystem condition, including water quality (dissolved oxygen, salinity, temperature, pH, nutrients and chlorophyll, suspended solids, fecal coliform bacteria and coliphages), sediment quality (granulometry, organic matter content, chemical contaminant concentrations), biological condition (diversity and abundance of benthic fauna, fish tissue contaminant levels and pathologies), and human dimensions (fish-tissue contaminant levels relative to human-health consumption limits, various aesthetic properties). Use of a probabilistic sampling design facilitated the calculation of statistics to estimate the spatial extent of the Reserve classified according to various categories (i.e., Good, Fair, Poor) of ecological condition relative to established thresholds of these indicators, where available. Overall, the majority of subtidal habitat in the SINERR appeared to be healthy, with over half (56.7 %) of the Reserve area having water quality, sediment quality, and benthic biological condition indicators rated in the healthy to intermediate range of corresponding guideline thresholds. None of the stations sampled had one or more indicators in all three categories rated as poor/degraded. While these results are encouraging, it should be noted that one or more indicators were rated as poor/degraded in at least one of the three categories over 40% of the Reserve study area, represented by 12 of the 30 stations sampled. Although measures of fish tissue chemical contamination were not included in any of the above estimates, a number of trace metals, pesticides, polybrominated diphenyl ethers (PBDEs), and polychlorinated biphenyls (PCBs) were found at low yet detectable levels in some fish at stations where fish were caught. Levels of mercury and total PCBs in some fish specimens fell within EPA guideline values considered safe, given a consumption rate of no more than four fish meals per month. Moreover, PCB congener profiles in sediments and fish in the SINERR exhibit a relative abundance of higher-chlorinated homologs which are uniquely characteristic of Aroclor 1268. It has been well-documented that sediments and fish in the creeks and marshes near the LCP Chemicals Superfund site, near Brunswick, Georgia, also display this congener pattern associated with Aroclor 1268, a highly chlorinated mixture of PCBs used extensively at a chlor-alkali plant that was in operation at the LCP site from 1955-1994. This report provides results suggesting that the protected habitats lying within the boundaries of the SINERR may be experiencing the effects of a legacy of chemical contamination at a site over 40km away. These effects, as well as other potential stressors associated with increased development of nearby coastal areas, underscore the importance of establishing baseline ecological conditions that can be used to track potential changes in the future and to guide management and stewardship of the otherwise relatively unspoiled ecosystems of the SINERR.

Using wildlife activity and antibiotic resistance analysis to model bacterial water quality in coastal ponds

Models that help predict fecal coliform bacteria (FCB) levels in environmental waters can be important tools for resource managers. In this study, we used animal activity along with antibiotic resistance analysis (ARA), land cover, and other variables to build models that predict bacteria levels in coastal ponds that discharge into an estuary. Photographic wildlife monitoring was used to estimate terrestrial and aquatic wildlife activity prior to sampling. Increased duck activity was an important predictor of increased FCB in coastal ponds. Terrestrial animals like deer and raccoon, although abundant, were not significant in our model. Various land cover types, rainfall, tide, solar irradiation, air temperature, and season parameters, in combination with duck activity, were significant predictors of increased FCB. It appears that tidal ponds allow for settling of bacteria under most conditions. We propose that these models can be used to test different development styles and wildlife management techniques to reduce bacterial loading into downstream shellfish harvesting and contact recreation areas.

Estudo da suscetibilidade aos antibióticos em isolados de Enterobacteriaceae provenientes de suiniculturas Portuguesas

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

Laterite as a disinfectant.

A groundwater programme monitoring flow and quality of a potable water spring in a slum district in Kampala, Uganda revealed that although latrines acted as the principal means of organic waste disposal for the 1000 plus people living in the spring’s catchment, levels of faecal indicator bacteria (TVC 45 Deg C) in spring discharge remained at or below detection during the dry season, despite the presence of high levels of chloride (45mg/l-56mg/l) and nitrate (23mg/l – 30mg/l NO3-N), indicating sewage impacts. A programme of column and batch testing of laterite underlying the area provided a means of investigating the soil’s attenuation capacity under more controlled conditions.
X-ray diffraction analyses revealed the laterite to be dominated by quartz and kaolinite with minor (<5% by volume) quantities of haematite. Batch studies revealed that over 99% of bacteriophage adsorbed to haematite in less than 5 minutes. By contrast batch tests on haematite-free soil samples from the Blue Hills in Australia showed that although they had comparable dominant mineralogy and iron coverage on their surfaces (determined from Energy dispersive X-ray fluorescence) they had negligible ability to adsorb H40/1.
Based on the results of the batch studies using natural soils, a programme of batch studies, undertaken using pure haematite showed the mineral to have an extremely high capacity to adsorb bacteriophage, and suggested that it was responsible for the levels of attenuation observed.
The results of column studies were in keeping with the findings of batch experiments. Injection of 20 pore volumes of 300 pfu/mL of the bacteriophage H40/1 into a 20mm diameter glass column packed with sand sized (Ø>500µm) laterite revealed that the column could irreversibly remove over 2.5 log10 bacteriophage over its 10cm length.
Importance:
Mineralogical and batch test data provide convincing evidence to show that laterite can potentially act as an inexpensive means of removing micro organisms from water. The material, particularly in nodular form, displays considerable potential to act as an alternative filter material to conventional quartz filter sands.

Geophysical and hydrogeological characterisation of the impacts of on-site wastewater treatment discharge to groundwater in a poorly productive bedrock acquifer

Contaminants discharging from on-site wastewater treatment systems (OSWTSs) can impact groundwater quality, threatening human health and surface water ecosystems. Risk of negative impacts becomes elevated in areas of extreme vulnerability with high water tables, where thin unsaturated intervals limit vadose zone attenuation. A combined geophysical/hydrogeological investigation into the effects of an OSWTS, located over a poorly productive aquifer (PPA) with thin subsoil cover, aimed to characterise effluent impacts on groundwater. Groundwater, sampled from piezometers down-gradient of the OSWTS percolation area displayed spatially erratic, yet temporally consistent, contaminant distributions. Electrical resistivity tomography identified an area of gross groundwater contamination close to the percolation area and, when combined with seismic refraction and water quality data, indicated that infiltrating effluent reaching the water table discharged to a deeper more permeable zone of weathered shale resting on more competent bedrock. Subsurface structure, defined by geophysics, indicated that elevated chemical and microbiological contaminant levels encountered in groundwater samples collected from piezometers, down-gradient of sampling points with lower contaminant levels, corresponded to those locations where piezometers were screened close to the weathered shale/competent rock interface; those immediately up-gradient were too shallow to intercept this interval, and thus the more impacted zone of the contaminant plume. Intermittent occurrence of faecal indicator bacteria more than 100 m down gradient of the percolation area suggested relatively short travel times. Study findings highlight the utility of geophysics as part of multidisciplinary investigations for OSWTS contaminant plume characterisation, while also demonstrating the capacity of effluent discharging to PPAs to impact groundwater quality at distance. Comparable geophysical responses observed in similar settings across Ireland suggest the phenomena observed in this study are more widespread than previously suspected.

‘Studies on Bacterial Indicators and Pathogen Vibrio parahaemolyticus in Cochin Backwater

Microbiological studies on the incidence, behaviour, activity and ecological implications of marine micro~organisms, particularly microbial pathogens in coastal waters and estuaries exhibit the increasing concern and awareness of environmental impacts on health and wealth. Marine microbiologists have been active in investigating on the distribution, kinds of organisms and their activity in the environment. However, informations on the effect of environment on the ecology or on the distribution (spatial/temporal) of microbial comunity and competition among groups inhabiting the ecosystem are sparE§L Estuarine environment are complex with respect to diversity of habitats, variation in physicochemical parameters and contamination by terrestrial bacterial species. Being the organisms of‘public health significance, ecological studies on total coliforms, faecal coliforms, faecal streptococci, §. ggli and X. parahaemolyticus have great relevance as studies of these types would provide a wealth of information to environmentalists and to fishery industry. In order to evalé%e the status, role and significance of potentially hazardous bacterial species in natural environment it is necessary to monitor the ecology of such organisms systematically in relation to physico-chemical parameters