Dynamics and interaction of organic carbon, turbidity and bacteria in a karst aquifer system

The dynamics of organic carbon (OC), turbidity, faecal indicator bacteria and physicochemical parameters was studied in a karst system near Yverdon, Switzerland. Online measurements and sampling were done at a swallow hole draining an agricultural surface (the input), and two groups of springs (the outputs) that often show bacterial contamination. A fluorescent tracer that was injected into the swallow hole during low-flow conditions first arrived at the springs 10–12 days after injection; the total recovery rate was 29%. Previous tracer tests during high-flow conditions gave shorter travel times. After a major rainfall event, a primary turbidity peak was observed at the springs. It coincides with the rising limb of the hydrograph, indicating remobilisation of autochthonous particles from the aquifer. A secondary turbidity peak occurs several days later, suggesting the arrival of allochthonous particles from the swallow hole. Wider peaks of OC and bacteria were observed simultaneously. Applying methods from molecular microbiology (PCR-DGGE) allowed characterisation of the bacterial communities at the swallow hole and the springs. The results demonstrate that the swallow hole is an important source of groundwater contamination, while its contribution to aquifer recharge is insignificant. OC appears to be a better indicator for bacterial contamination than turbidity.     

沉积物和悬浮颗粒物在岩溶水系统污染物衰减与运移中的作用

本文讨论了沉积物和悬浮颗粒物/胶体在岩溶水系统污染物衰减与运移中的作用,介绍了关于瑞士某岩溶水系统内颗粒物与微生物污染间关系的实验结果。该实验场包括一个接纳农田排水的落水洞及与其相连的两个泉,其中一个泉是供水水源地。利用便携式粒子计实时监测泉水中颗粒分布的变化情况,同时对浊度、总有机碳和其它参数进行连续监测;并对水中E型大肠菌群进行了分析。在暴雨过后,观测到了浊度的两种不同表现:即泉排泄量增加时,浊度最初表现为脉状水动力条件下岩溶系统内沉积物的二次运移;几天后观测到的浊度则表明水已从落水洞到了泉口,常常具有高的总有机碳、硝酸盐及E型大肠菌群。颗粒分布分析结果差异更大:早期的浊度是不同粒径颗粒物的混合,而后期的浊度更多是由较细颗粒构成,因为大颗粒物已沉积在落水洞到泉口之间的岩溶系统内。E型大肠菌群与0.9～1.5 μm颗粒物之间具有很好的相关性。因而较细颗粒的相对增加可应用于微生物污染 “早期预报系统”。除了水污染,颗粒分还可以帮助我们研究岩溶环境的其它问题,如土壤侵蚀。      

Microbiological monitoring and classification of karst springs

The use of groundwater as a drinking water resource requires knowledge of its microbiological status and quality. In contrast to conventional microbiological monitoring of groundwater, the present study not only considers faecal indicator bacteria, but also covers a wide spectrum of microorganisms, including bacterial pathogens (verotoxin-producing E. coli, Campylobacter spp. and Salmonella spp., as well as Pseudomonas aeruginosa), human enteric viruses (norovirus, enterovirus, rotavirus and adenovirus) and parasitic protozoa (Cryptosporidium oocysts and Giardia cysts). Samples collected at karst sites of the Swiss National Groundwater Monitoring network revealed the presence of a large diversity of microorganisms of faecal origin, the occurrence of which could be linked to specific hydrogeological settings and situations. The findings represent a ‘snapshot’ of the microbiological status at the monitoring sites and provide a national overview of the types and presence of microorganisms in Swiss karst groundwater. In addition to microbiological parameters related to faecal contamination, the overall bacterial load in groundwater was assessed using cell density measurements (i.e. total cell count), which yielded typical ranges for this ecological parameter. The study highlights differential vulnerability of karst groundwater to microbiological contamination, as well as its relationship with the microbial biocenoses, i.e. the interplay of allochthonous and autochthonous microbial components. On the basis of this data set, a microbiological classification of karst aquifers is proposed and discussed with respect to spring dynamics and vulnerability.     

Using turbidity dynamics and geochemical variability as a tool for understanding the behavior and vulnerability of a karst aquifer

In a karst system, the characterization of transport properties is based on the comparison of natural tracers observed at the inlet (a swallow hole on the karst plateau) and the outlets of the system (a spring and a well). At Norville, northwest France, electrical conductivity (EC) and turbidity (T) were used as natural tracers for characterizing dissolved elements (surface water geochemistry) and particulate matter transport, respectively. Two methods were used for this study: (1) a comparison of the relations between EC, T and spring discharge (Q) by means of normalized EC–T–Q curves, and (2) a principal component analysis (PCA) including water geochemistry data in addition to EC, T and Q. Three different characteristic flood events have been chosen for the analyses. EC–T–Q curves highlighted the direct transfer, resuspension and deposition of particles during their transport in the karst network. Transport from the swallow hole to both the spring and the well appeared to be dominated by karst-conduit flow. On the other hand, PCA results showed a diffuse source of contamination of groundwaters by nitrate and a point-source contamination of groundwaters by streaming/runoff surface waters with high turbidity and phosphate concentration infiltrated at the swallow hole.     

Aquifer Discharge Drives Microbial Community Change in Karst Estuaries

Karst estuaries are unique systems governed by freshwater inputs that flow directly to the sea through karst conduits and/or matrices. Because they are found at the interface between terrestrial and marine environments, they can act as sentinels to climate change and as indicators of aquifer health. The physical and geochemical gradients formed in karst estuaries are a direct result of the interactions between inland hydrological conditions and sea level. To examine the influence these gradients have on microbial communities in the water column and sediment of a karst estuary, we studied the spring-fed Double Keyhole Karst Estuary in west central Florida for a 2-year period. Four sites were monitored within this system starting at the Double Keyhole Spring Conduit extending 2 km west through the estuary toward the Gulf of Mexico. Water column and sediment samples were collected quarterly at all sites from September 2011 through September 2013. Archaeal, bacterial, and microbial eukaryote communities were analyzed using quantitative PCR and length-heterogeneity PCR. The biological communities were analyzed in the context of hydrological, physical, and geochemical parameters in order to determine which factor(s) governed the observed changes of microbial abundance and richness. The major finding of this study was that microbial community change in this karst estuary was primarily driven by the volume of aquifer discharge and associated physical gradients, and less by the geochemical fluctuations within the system.     

Molecular characterization of dissolved organic matter associated with the Greenland ice sheet

Subsurface microbial oxidation of overridden soils and vegetation beneath glaciers and ice sheets may affect global carbon budgets on glacial-interglacial timescales. The likelihood and magnitude of this process depends on the chemical nature and reactivity of the subglacial organic carbon stores. We examined the composition of carbon pools associated with different regions of the Greenland ice sheet (subglacial, supraglacial, proglacial) in order to elucidate the type of dissolved organic matter (DOM) present in the subglacial discharge over a melt season. Electrospray ionization (ESI) Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry coupled to multivariate statistics permitted unprecedented molecular level characterization of this material and revealed that carbon pools associated with discrete glacial regions are comprised of different compound classes. Specifically, a larger proportion of protein-like compounds were observed in the supraglacial samples and in the early melt season (spring) subglacial discharge. In contrast, the late melt season (summer) subglacial discharge contained a greater fraction of lignin-like and other material presumably derived from underlying vegetation and soil. These results suggest (1) that the majority of supraglacial DOM originates from autochthonous microbial processes on the ice sheet surface, (2) that the subglacial DOM contains allochthonous carbon derived from overridden soils and vegetation as well as autochthonous carbon derived from in situ microbial metabolism, and (3) that the relative contribution of allochthonous and autochthonous material in subglacial discharge varies during the melt season. These conclusions are consistent with the hypothesis that, given sufficient time (e.g., overwinter storage), resident subglacial microbial communities may oxidize terrestrial material beneath the Greenland ice sheet.     

Sources and composition of organic matter for bacterial growth in a large European river floodplain system (Danube, Austria)

Dissolved and particulate organic matter (DOM and POM) distribution, lignin phenol signatures, bulk elemental compositions, fluorescence indices and microbial plankton (algae, bacteria, viruses) in a temperate river floodplain system were monitored from January to November 2003. We aimed to elucidate the sources and compositions of allochthonous and autochthonous organic matter (OM) in the main channel and a representative backwater in relation to the hydrological regime. Additionally, bacterial secondary production was measured to evaluate the impact of organic carbon source on heterotrophic prokaryotic productivity. OM properties in the backwater tended to diverge from those in the main channel during phases without surface water connectivity; this was likely enhanced due to the exceptionally low river discharge in 2003. The terrestrial OM in this river floodplain system was largely derived from angiosperm leaves and grasses, as indicated by the lignin phenol composition. The lignin signatures exhibited significant seasonal changes, comparable to the seasonality of plankton-derived material. Microbially-derived material contributed significantly to POM and DOM, especially during periods of low discharge. High rates of bacterial secondary production (up to 135 μg C L(-1) d(-1)) followed algal blooms and suggested that autochthonous OM significantly supported heterotrophic microbial productivity.     

Catastrophic flood in the polje of Cetinje in February 1986, a typical example of the environmental impact of Karst

The catastrophic flood of February 1986 in the polje of Cetinje, unrecorded in the 500-year long written history of the old Montenegrin capital, was an opportunity to examine some specific features of the complex hydrogeologic system. Above all, it is the incompatibility of the epikarst and endokarst systems of karst aquifers, collectors-conductors, and groundwater reservoirs that causes a hydrological environmental impact on the polje in extremely wet (autumnwinter) periods of the year. This year's flood has indicated that this epikarst system, generally dry most of the year, has a relatively large water-receiving capacity, and that the deeper endokarst system has all attributes of a karst aquifer, much variable transmissivity and low storage capacity.     

Contribution of allochthonous carbon to American shad production in the Mattaponi River,Virginia, using stable isotopes

Our objective was to quantify the contribution of autochthonous, locally-produced phytoplankton, and allochthonous, terrestrial-derived organic matter (OM) to the production of young-of-year (YOY) American shad(Alosa sapidissima) using stable isotopes. We measured the carbon and nitrogen stable isotope composition of YOY American shad in the tidal fresh water of the Mattaponi River, a tributary in the York River estuary, during three consecutive years. The isotopic ratios of larval American shad varied among years, indicating a switch from reliance on a primarily autochthonous food web pathway during low and moderate discharge years (50–90%; 2002, 2004) to a primarily allochthonous pathway during a high discharge year (< 35% phytoplankton; 2003). Reliance on phytoplankton by larval fish declined exponentially with increasing Mattaponi River discharge. In 2003, juvenile production was also supported by allochthonous OM, though autochthonous phytoplankton accounted for an increasingly large fraction during June through August, up to 40–55%. We also found a long-term, positive relationship between the duration of above average flow during April through June in the Mattaponi River and a corresponding index of juvenile American shad abundance. The largest American shad cohort recorded since 1967 was observed in 2003, a high discharge year. The production of this cohort was largely supported by allochthonous OM. The results suggest an important link between river discharge, energy flow, and recruitment, wherein high discharge favors reliance on terrestrial carbon by YOY American shad, owing to changes in zooplankton diet, macroinvertebrate abundance, or both, and also favors high American shad abundance.     

Lipids of Recent sediments,Part I: Straight-chain hydrocarbons and carboxylic acids of some temperate lacustrine and sub-tropical lagoonal/tidal flat sediments

Analyses of the lipid constituents of three temperate lacustrine sediments (Rostherne Mere, Esthwaite Water and Grasmere, Great Britain) and three sub-tropical lagoonal/tidal flat sediments (U.S. Virgin Islands, Rockhampton, Qld., Australia and Port Moresby, Papua, New Guinea) indicate the presence of straight-chain components attributable to autochthonous and allochthonous sources. Despite broad similarities in carbon-number distributions, minor differences observed in the respective hydrocarbon and in the respective fatty-acid compositions of the various sediments reflect differences in the sedimentary environments. Lipid constituents obtained by organic solvent extraction following acid hydrolysis appear to include a larger bacterial contribution than that observed in the lipids extracted by solvent alone.     

Comparison of microbial communities in petroleum-contaminated groundwater using genetic and metabolic profiles at Kyonggi-Do,South Korea

This research has been performed to determine the differences in microbial communities according to physicochemical properties such as concentrations of volatile aromatic hydrocarbons such as benzene, toluene, ethylbenzene, and xylene (BTEX), dissolved oxygen (DO), electron acceptors, etc., in oil-contaminated groundwaters at Kyonggi-Do, South Korea. The properties of bacterial and microbial communities were analyzed by 16S polymerase chain reaction (PCR) denaturing gradient gel electrophoresis (DGGE) fingerprinting method and community-level physiological profiling (CLPP) using Eco-plate, respectively. Based on the DGGE fingerprints, the similarities of bacterial community structures were high with similar DO levels, and low with different DO levels. Whereas the dominant bacterial groups in GW13 (highest BTEX and lowest DO) were acidobacteria, α-proteobacteria, β-proteobacteria, γ-proteobacteria, δ-proteobacteria, and spirochetes, those in GW7 (highest BTEX and highest DO) were actinobacteria, α-proteobacteria, β-proteobacteria, γ-proteobacteria, δ-proteobacteria, and sphingobacteria. Based on the CLPP results, the groundwater samples were roughly divided into three groups: above 4 mg/L in DO (group 1: GW3 and GW7), below 4 mg/L in DO (group 2: GW8, W1, W2, W3, and BH10), and highly contaminated with BTEX (group 3: GW13). Shannon index showed that the microbial diversities and equitabilities were higher in shallower aquifer samples. Overall, this study verified that the greatest influencing factors on microbial/bacterial communities in groundwaters were DO and carbon sources, although BTEX concentration was one of the major factors.     

Spatial-seasonal dynamics of chromophoric dissolved organic matter in Lake Taihu, a large eutrophic, shallow lake in China

From 2005 to 2009, the spatial distribution and the seasonal dynamics of chromophoric dissolved organic matter (CDOM) were explored in Lake Taihu in eastern China. The spatial-seasonal dynamics of CDOM absorption and three CDOM composition variables, including spectral slope (S), spectral slope ratio (S_R) and the M value, defined as the ratio of absorption at 250 nm/365 nm, were analyzed and discussed. Furthermore, river input processes and degradation of phytoplankton were studied to assess their impact on CDOM composition and the factors involved in the spatial-seasonal variability of CDOM. The CDOM absorption coefficient at a wavelength of 350 nm, a(350), ranged from 1.37-9.55 m⁻¹ with a mean of 3.33 ± 1.32 m⁻¹. Spatially, higher a(350) values, but lower spectral slope, spectral slope ratio and M values, were recorded in the northern algae dominated bays while lower values were recorded in southeastern macrophyte dominated bays. The a(350) was significantly higher in inshore waters than in offshore waters. Values of S, S_R and M decreased during the flood and algal bloom season in spring and summer whereas a(350) increased. In the three river profiles, the gradual decrease of a(350) along the trajectory from the river mouth into the lake during the flood season showed the contribution of allochthonous CDOM. A laboratory phytoplankton degradation experiment was conducted to determine the contribution of CDOM production from phytoplankton. The significant increase of a(350) with time in the CDOM production experiment underlines the importance of autochthonous CDOM production during the algal bloom season. In summary, the significant increase of a(350) in spring and summer (algal bloom season) may be due to both the allochthonous CDOM input from the surrounding rivers and the autochthonous production of CDOM from degrading phytoplankton.     

Increased Variability of Microbial Communities in Restored Salt Marshes nearly 30?Years After Tidal Flow Restoration

We analyzed microbial diversity and community composition from four salt marsh sites that were impounded for 40?C50?years and subsequently restored and four unimpounded sites in southeastern Connecticut over one growing season. Community composition and diversity were assessed by terminal restriction fragment length polymorphism (TRFLP) and sequence analysis of 16S ribosomal RNA (rRNA) genes. Our results indicated diverse communities, with sequences representing 14 different bacterial divisions. Proteobacteria, Bacteroidetes, and Planctomycetes dominated clone libraries from both restored and unimpounded sites. Multivariate analysis of the TRFLP data suggest significant site, sample date, and restoration status effects, but the exact causes of these effects are not clear. Composition of clone libraries and abundance of bacterial 16S rRNA genes were not significantly different between restored sites and unimpounded sites, but restored sites showed greater temporal and spatial variability of bacterial communities based on TRFLP profiles compared with unimpounded sites, and variability was greatest at sites more recently restored. In summary, our study suggests there may be long-lasting effects on stability of bacterial communities in restored salt marshes and raises questions about the resilience and ultimate recovery of the communities after chronic disturbance.     

Contamination risk of the Alburni Karst System (Southern Italy)

The Alburni massif (1742m a.s.l.) stretches NW–SE, about 23km long and 9–10km wide, covering 246km² with an average elevation of about 940m a.s.l. This massif, with more than 500 caves, is the most important karst area in southern Italy. The karst channel network is hierarchically organized: some channels feed a major spring (1m³/s) with a very short transit time while others communicate directly with the basal water table related to other springs (Q > 3m³/s).There are several dolines and swallow holes just above the basal water table and in the urbanized areas; for years a swallow hole directly transferred pollutants into the aquifer. The contamination vulnerability map shows that the prevalent vulnerability degree ranges from high to very high, due to the widespread karstification of the area and to the presence, on the plateau, of large vegetated areas with gentle slopes favouring fast infiltration.Hence it is important to ascertain the human impact on the area and the consequent contamination risk of the aquifer of the Alburni karst area. Three main layers were created to assess groundwater contamination risk: the vulnerability map, the hazard map, and the value map.The groundwater contamination risk map stresses the importance in a park area of aquifer vulnerability, which strongly influences the risk: indeed, the prevalent moderate degree of risk in the final map depends on the high vulnerability and the low hazard degree. However, in the future it is crucial to take into account the nature of the agricultural land use allowed in the park, which could increase the hazard degree and consequently the risk degree.     

岩溶泉域降雨径流水文过程的模拟——以重庆金佛山水房泉为例

通过地貌、水文地质、土壤、植被、地下洞穴管道等实地调查,以及示踪试验确定水房泉泉域范围和地下水文系统特征,并通过代表性点的土壤入渗试验、降雨和流量监测以及DEM数据等获得泉域水文模型所需的面积、结点高程、入渗率、糙率、管道长度、含水层孔隙度、出流系数等参数。选择SCS径流曲线模型估算地表产流,利用SWMM模型模拟泉域对场降雨的径流响应过程。通过运行模型,与实测流量比较,结果显示模拟曲线与观测流量曲线吻合较好,用于校正和验证的两场降雨产流的模拟误差分别为9.5%和12%,表明SWMM可应用于岩溶区以管道流为主要排水系统的含水介质的模拟。     

Factors controlling the distribution of anaerobic methanotrophic communities in marine environments: Evidence from intact polar membrane lipids

Three distinct types of microbial consortia appear to mediate the anaerobic oxidation of methane with sulfate as electron acceptor in marine sediments and are distributed ubiquitously. These consortia consist of ANerobic MEthanotrophic (ANME) archaea of the ANME-1, ANME-2 and ANME-3 clades and their sulfate-reducing bacterial partners either of the Desulfosarcina-Desulfococcus (ANME-1/DSS and ANME-2/DSS) or Desulfobulbus spp. (ANME-3/DBB) branches. Frequently one consortium type dominates the community, but the selective factors are not well constrained. Here we analyzed patterns in the composition of intact polar lipids extracted from bacterial and archaeal communities of different marine seep environments. Further, we investigated if different environmental and geographical factors were responsible for the observed patterns, and hence could be important in the selection of seep communities. Intact polar lipids (IPLs) provide a more robust distinction of the composition of extant communities than their less polar derivatives. In ANME-1/DSS-dominated communities, glycosidic- and phospho-glyceroldialkylglyceroltetraethers were abundant, while ANME-2/DSS and ANME-3/DBB-dominated communities showed abundant archaeol-based IPLs, either with glycosidic and phospho-headgroups or only phospho-headgroups, respectively. The relative proportion of bacterial IPLs varied widely from 0% to 93% and was generally lower in samples of the ANME-1 type, suggesting lower bacterial biomasses in the respective communities. In addition to these lipid signatures, distinctive features were related to the habitat characteristics of these communities: lower amounts of phosphate-based IPLs were generally observed in communities from calcified microbial mats compared to sediments, which may reflect phosphate limitation. Based on statistical analyses of IPLs and environmental data this study constrained for the first time the occurrence of three environmental factors controlling the distribution of different ANME-associated communities in a wide range of hydrocarbon seep systems. Habitats dominated by ANME-1/DSS communities were characterized by high temperature and low oxygen content in bottom waters (or even anoxia), while ANME-2/DSS and ANME-3/DBB-dominated sediments were located in settings with lower temperatures and higher oxygen content in bottom waters. Furthermore, ANME-2/DSS communities were particularly prominent in environments in which a relatively high supply of sulfate was sustained.     

Water quality deterioration at a karst spring (Gallusquelle,Germany) due to combined sewer overflow: evidence of bacterial and micro-pollutant contamination

The concurrent use of karst aquifers as drinking water resources and receptors of combined sewer overflow lacking appropriate pre-treatment may cause conflicts between drinking water supply and storm water management. A storm water tank (SWT) for combined wastewater is identified as the source of sporadic contamination of a karst spring (Gallusquelle, “Schwäbische Alb”, SW Germany) used for public water supply. Spring water quality was examined by routine and event sampling and by evaluating physicochemical and microbiological parameters. The total number of microbial colonies growing at 20°C and the number of Escherichia coli colonies rose to values up to four orders of magnitude higher than background, 2–5 days after overflow of the SWT. High concentrations of chloride, sodium, and total organic carbon (TOC) and high values of turbidity coincide with this increase. However, high bacterial contamination is also observed while turbidity and TOC are low. Several wastewater-related organic micro-pollutants such as chlorinated and non-chlorinated organophosphates were detected in the SWT and, depending on their K_ow values and their biodegradability, in lower concentrations at the spring.     

Extractable and bound lipid components in a freshwater sediment

Lipids directly extracted by organic solvents from sediment of a small eutrophic lake (Crose Mere) comprise free hydrocarbons, alcohols, sterols and monocarboxylic acids together with an esterified component yielding additional amounts of alcohols, sterols and monocarboxylic acids on saponification. The carbon-number distributions of the free lipid compound classes indicate a major contribution from allochthonous sources whereas the distributions within compound classes released on saponification are attributable to autochthonous sources. The more advanced state of decomposition of terrestrial biota relative to aquatic biota and the composition of the source organisms are postulated to be factors leading to the differences in distribution of corresponding free and esterified lipid classes. Acid hydrolysis of solvent-extracted sediment liberates additional lipid constituents belonging to the above compound classes. Of these bound lipids, the n-alkanes and branched/cyclic alkanoic acids show a more significant bacterial contribution than do the corresponding components obtained by solvent extraction alone.     

Environmental factors controlling the spatiotemporal distribution of microbial communities in a coastal, sandy aquifer system (Doñana, southwest Spain)

Currently, aquifers are considered to be ecosystems that interchange materials and energy with other systems located in their surroundings. The aquifer system of Doñana (southwest Spain) has been studied over recent decades from a hydrogeological point of view, although nothing is known about its biological or ecological aspects. In order to describe the general characteristics of its microbial communities, bacterial abundance, cell biomass, bacterial biomass and microbial activities of functional groups were investigated by sampling, over a 2-year period, 13 wells located in the vicinity of four very productive shallow lakes in the most superficial part of this coastal, sandy aquifer system. Multivariate analysis of variance (MANOVA) indicated differences in abundance and biomass variables among sampling locations, seasons and sampling locations × seasons. Principal component analysis showed that temperature and dissolved oxygen appeared to be the most important factors controlling the temporal variability of microbial communities. Hydrological connectivity between surface water and groundwater was important in the control of the spatiotemporal distribution of microbial communities. Due to this hydrological connection, the aquifer system and the wetlands constitute a unique entity, a unique ecosystem, called the “hydroecosystem”, where microbial communities could play a central ecological role.     

Spring flow,bacterial contamination,and water resources in rural Haiti

Field data from 25 karst springs were collected during the summer of 2008 near Verrettes, Haiti, to gain a better understanding of water quality in fresh water springs used as the primary source of potable water in rural Haiti. Two water samples were taken at each spring for bacterial analysis: one sample was submitted to a local hospital for analysis, while a duplicate sample was cultured using inexpensive and commercially available Coliscan^® Easygel^® Kits. Both capped and uncapped springs were sampled in order to evaluate whether capping improves water quality. Water quality parameters and flow volume estimates were recorded at each spring; however, no correlations between field water parameters and the presence of E. coli or total coliform bacteria were found. Spring flow rates varied from 0.4 to 268.5 L/min. Geologic contacts and regional structures trend in a northwest direction and appear to be exerting a primary control on spring locations. E. coli and total coliform colony counts from the local hospital and Coliscan^® Easygel^® Kits yielded different results for the same springs due to sample handling and analysis complications common in rural Haiti. Water sample analysis and storage problems encountered during this study likely resulted in underprediction, rather than overprediction, of bacterial contamination. Water from 71 to 100% of the springs was unsafe to drink based on the World Health Organization (WHO) drinking water standard. Both capped and uncapped springs had bacterial counts in excess of the WHO standard, suggesting that water treatment from all sources is necessary to ensure clean and safe drinking water. Negatively impacted subterranean microbial ecosystems, poor sanitation practices, shallow karst aquifers with open flow paths, and high spring water temperatures, averaging 26.5°C, may be contributing to the observed bacterial abundance.