Longitudinal Study of Microbial Diversity and Seasonality in the Mexico City Metropolitan Area Water Supply System

In the Mexico City metropolitan area (MCMA), 70% of the water for 18 million inhabitants is derived from the Basin of Mexico regional aquifer. To provide an overview of the quality of the groundwater, a longitudinal study was conducted, in which 30 sites were randomly selected from 1,575 registered extraction wells. Samples were taken before and after chlorine disinfection during both the rainy and dry seasons (2000-2001). Microbiological parameters (total coliforms, fecal coliforms, streptococci, and Vibrio spp.), the presence of Helicobacter pylori, and physicochemical parameters, including the amount of trihalomethanes (THMs), were determined. Although microorganisms and inorganic and organic compounds were evident, they did not exceed current permissible limits. Chlorine levels were low, and the bacterial counts were not affected by chlorine disinfection. Eighty-four bacterial species from nine genera normally associated with fecal contamination were identified in water samples. H. pylori was detected in at least 10% of the studied samples. About 40% of the samples surpassed the THM concentration allowed by Mexican and U.S. regulations, with levels of chloroform being high. The quality of the water distributed to the MCMA varied between the rainy and dry seasons, with higher levels of pH, nitrates, chloroform, bromodichloromethane, total organic carbon, and fecal streptococci during the dry season. This study showed that the groundwater distribution system is susceptible to contamination and that there is a need for a strict, year-round disinfection strategy to ensure adequate drinking-water quality. This situation in one of the world's megacities may reflect what is happening in large urban centers in developing countries which rely on a groundwater supply.     

Microbiological implications of periurban agriculture and water reuse in Mexico City

Background

Recycled treated or untreated wastewater represents an important health challenge in developing countries due to potential water related microbiological exposure. Our aim was to assess water quality and health implications in a Mexico City periurban agricultural area.

Methodology/Principal Findings

A longitudinal study in the Xochimilco wetland area was conducted, and 42 sites were randomly selected from 211, including irrigation water canals and effluents of treatment plants. Sample collection took place during rainy and dry seasons (2000–2001). Microbiological parameters (total coliforms, fecal coliforms, streptococci/enterococci, and bacteria other than Vibrio grown on TCBS), Helicobacter pylori, and physicochemical parameters including trihalomethanes (THM) were determined. Fecal coliforms and fecal streptococci are appropriate indicators of human or animal fecal contamination. Fecal coliform counts surpass Mexican and World Health Organization irrigation water guidelines. Identified microorganisms associated with various pathologies in humans and domestic animals comprise Escherichia coli, Klebsiella spp., Salmonella spp., Enterobacter spp., Enterococcus spp., and Pseudomonas spp; H. pylori was also present in the water. An environmental characteristic of the canal system showed high Total Organic Carbon content and relatively low dissolved oxygen concentration; residual chlorine as a disinfection control is not efficient, but THMs do not represent a problem. During the rainy season, temperature and conductivity were higher; in contrast, pH, dissolved oxygen, ammonia, and residual chlorine were lower. This is related with the continuous load of feces from human and animal sources, and to the aquatic systems, which vary seasonally and exhibit evidence of lower water quality in effluents from treatment plants.

Conclusions/Significance

There is a need for improvement of wastewater treatment systems, as well as more efficient monitoring, regulation, and enforcement procedures for wastewater disposal into bodies of water.     

Microcystin in plants that treat water for human consumption in a tropical environment: the Metropolitan Area of Costa Rica

We measured microcystin levels in water of the Metropolitan Area of Costa Rica by competitive inhibition ELISA and we quantified total coliforms, fecal coliforms, Escherichia coli (by a Most Probable Number method) and aerobic count. We wanted to identify any cyanotoxin correlation with these parameters, as a public health risk. We sampled in the rainy season of 2003 (April-October) and in the dry season of 2004 (February-March) (30 samples/season). We sampled pre-treated, semi-treated and treated water. Microcystin levels < 0.5 ppb were found in the rainy season (and > 0.5 ppb in the dry season). Dry season levels exceeded World Health Organization limits (1.0 ppb). Cyanotoxins occurred in the Tres Rios plant. We did not find a correlation between these microbiologic parameters of water quality and microcystin levels in water.     

Distribution and Significance of Fecal Indicator Organisms in the Upper Chesapeake Bay

Total viable aerobic, heterotrophic bacteria, total coliforms, fecal coliforms, and fecal streptococci were enumerated in samples collected at five stations located in the Upper Chesapeake Bay, December 1973 through December 1974. Significant levels of pollution indicator organisms were detected at all of the stations sampled. Highest counts were observed in samples collected at the confluence of the Susquehanna River and the Chesapeake Bay. The indicator organisms examined were observed to be quantitatively distributed independently of temperature and salinity. Counts were not found to be correlated with concentration of suspended sediment. However, significant proportions of both the total viable bacteria (53%) and fecal indicator organisms (>80%) were directly associated with suspended sediments. Correlation coefficients (r) for the indicator organisms examined in this study ranged from r = 0.80 to r = 0.99 for bottom water and suspended sediment, respectively. Prolonged survival of fecal streptococci in most of the sediment samples was observed, with concomitant reduction of the correlation coefficient from r = 0.99, fecal streptococci to total coliforms in water, to r = 0.01, fecal streptococci to fecal coliforms in sediments. The results of this study compared favorably with fecal coliforms: fecal streptococci ratios for the various sample types. Characterization of organisms beyond the confirmed most-probable-number procedure provided good correlation between bacterial indicator groups.     

Comparison of bacterial indicators and sampling programs for drinking water systems

A total of 48 French villages were selected for a survey of water quality from February 1983 to June 1984 as part of an epidemiological study conducted in the rural area of the Rh?ne-Alpes region. Water samples were collected and analyzed on a weekly basis in each village. Bacteriological analysis of each water sample included enumeration of standard plate count bacteria, total and thermotolerant coliforms, and fecal streptococci. The water quality regulations are examined as to the analysis frequency, the volume of samples, and the relationship between the various bacterial indicators. Analyzing 300-ml, instead of 100-ml, samples tends to generate better information on single water samples. However, if many samples are analyzed over time from the same community, the value of using large volumes of water samples is diminished. The comparisons between bacterial indicators showed that the information obtained from the various indicators was very similar. However, fecal streptococci had a better predictive value of a negative test than coliforms with respect to the French standards.     

Comparison of bacterial indicators and sampling programs for drinking water systems.

A total of 48 French villages were selected for a survey of water quality from February 1983 to June 1984 as part of an epidemiological study conducted in the rural area of the Rhône-Alpes region. Water samples were collected and analyzed on a weekly basis in each village. Bacteriological analysis of each water sample included enumeration of standard plate count bacteria, total and thermotolerant coliforms, and fecal streptococci. The water quality regulations are examined as to the analysis frequency, the volume of samples, and the relationship between the various bacterial indicators. Analyzing 300-ml, instead of 100-ml, samples tends to generate better information on single water samples. However, if many samples are analyzed over time from the same community, the value of using large volumes of water samples is diminished. The comparisons between bacterial indicators showed that the information obtained from the various indicators was very similar. However, fecal streptococci had a better predictive value of a negative test than coliforms with respect to the French standards.     

Virus occurrence in municipal groundwater sources in Quebec, Canada

A 1 year study was undertaken on groundwater that was a source of drinking water in the province of Quebec, Canada. Twelve municipal wells (raw water) were sampled monthly during a 1 year period, for a total of 160 samples. Using historic data, the 12 sites were categorized into 3 groups: group A (no known contamination), group B (sporadically contaminated by total coliforms), and group C (historic and continuous contamination by total coliforms and (or) fecal coliforms). Bacterial indicators (total coliform, Escherichia coli, enteroccoci), viral indicators (somatic and male-specific coliphages), total culturable human enteric viruses, and noroviruses were analyzed at every sampling site. Total coliforms were the best indicator of microbial degradation, and coliform bacteria were always present at the same time as human enteric viruses. Two samples contained human enteric viruses but no fecal pollution indicators (E. coli, enterococci, or coliphages), suggesting the limited value of these microorganisms in predicting the presence of human enteric viruses in groundwater. Our results underline the value of historic data in assessing the vulnerability of a well on the basis of raw water quality and in detecting degradation of the source. This project allowed us to characterize the microbiologic and virologic quality of groundwater used as municipal drinking water sources in Quebec.     

Structure of bacterial communities in diverse freshwater habitats

The structures and dynamics of bacterial communities from raw source water, groundwater, and drinking water before and after filtration were studied in four seasons of a year, with culture-independent methods. Genomic DNA from water samples was analyzed by the polymerase chain reaction?- denaturing gradient gel electrophoresis system and by cloning of the 16S rRNA gene. Water samples exhibited complex denaturing gradient gel electrophoresis genetic profiles composed of many bands, corresponding to a great variety of bacterial taxa. The bacterial communities of different seasons from the four sampling sites clustered into two major groups: (i) water before and after filtration, and (ii) source water and groundwater. Phylogenetic analyses of the clones from the autumn sampling revealed 13 phyla, 19 classes, and 155 operational taxonomic units. Of the clones, 66% showed less than 97% similarities to known bacterial species. Representatives of the phyla Proteobacteria, Bacteroidetes, and Actinobacteria were found at all four sampling sites. Species belonging to the phylum Firmicutes were an important component of the microbial community in filtered water. Representatives of Enterobacteriaceae were not detected, indicating the absence of fecal pollution in the drinking water. Differences were found in the bacterial populations that were sampled from the same sites in different seasons. Each water habitat had a unique bacterial profile. Drinking water harbors diverse and dynamic microbial communities, part of which may be active and resilient to chlorine disinfection. This study provides, for the first time, basic data for uncultivable drinking water bacteria in Israel.     

Microbial contamination of vegetable crop and soil profile in arid regions under controlled application of domestic wastewater

Increasing lack of potable water in arid countries leads to the use of treated wastewater for crop production. However, the use of inappropriate irrigation practices could result in a serious contamination risk to plants, soils, and groundwater with sewage water. This research was initiated in view to the increasing danger of vegetable crops and groundwater contamination with pathogenic bacteria due to wastewater land application. The research was designed to study: (1) the effect of treated wastewater irrigation on the yield and microbial contamination of the radish plant under field conditions; (2) contamination of the agricultural soil profile with fecal coliform bacteria. Effluent from a domestic wastewater treatment plant (100%) in Jeddah city, Saudi Arabia, was diluted to 80% and 40% with the groundwater of the experimental site constituting three different water qualities plus groundwater as control. Radish plant was grown in two consecutive seasons under two drip irrigation systems and four irrigation water qualities. Upon harvesting, plant weight per ha, total bacterial, fecal coliform, fecal streptococci were detected per 100 g of dry matter and compared with the control. The soil profile was also sampled at an equal distance of 3 cm from soil surface for fecal coliform detection. The results indicated that the yield increased significantly under the subsurface irrigation system and the control water quality compared to surface irrigation system and other water qualities. There was a considerable drop in the count of all bacteria species under the subsurface irrigation system compared to surface irrigation. The bacterial count/g of the plant shoot system increased as the percentage of wastewater in the irrigation water increased. Most of the fecal coliform bacteria were deposited in the first few centimeters below the column inlet and the profile exponentially decreased with increasing depth.     

Validity of fecal coliforms, total coliforms, and fecal streptococci as indicators of viruses in chlorinated primary sewage effluents.

Quantities of combined chlorine that usually destroyed more than 99.999% of the indigenous fecal coliforms, total coliforms, and fecal streptococci in primary sewage effluents destroyed only 85 to 99% of the indigenous viruses present. Viruses were recovered from five of eight chlorinated primary effluents from which fecal coliforms were not recovered by standard most-probable-number procedures. The limited volumes of such chlorinated effluents that can be tested for indicator bacteria with currently available multiple-tube and membrane filter techniques restrict the value of fecal coliforms, fecal streptococci, and even total coliforms as indicators of viruses in these effluents. Although fecal coliforms and fecal streptococci are useful indicators of viruses in effluents from which these bacteria are recovered, the absence of these bacteria and even total coliforms from disinfected effluents (in standard tests) does not assure that viruses are also absent.     

Validity of fecal coliforms, total coliforms, and fecal streptococci as indicators of viruses in chlorinated primary sewage effluents.

Quantities of combined chlorine that usually destroyed more than 99.999% of the indigenous fecal coliforms, total coliforms, and fecal streptococci in primary sewage effluents destroyed only 85 to 99% of the indigenous viruses present. Viruses were recovered from five of eight chlorinated primary effluents from which fecal coliforms were not recovered by standard most-probable-number procedures. The limited volumes of such chlorinated effluents that can be tested for indicator bacteria with currently available multiple-tube and membrane filter techniques restrict the value of fecal coliforms, fecal streptococci, and even total coliforms as indicators of viruses in these effluents. Although fecal coliforms and fecal streptococci are useful indicators of viruses in effluents from which these bacteria are recovered, the absence of these bacteria and even total coliforms from disinfected effluents (in standard tests) does not assure that viruses are also absent.     

Detection of Acinetobacter spp. in rural drinking water supplies.

A bacteriological survey was conducted of untreated, individual groundwater supplies in Preston County, W.Va. Nearly 60% of the water supplies contained total coliforms in excess of the U.S. Environmental Protection Agency maximum contaminant level of 1 CFU/100 ml. Approximately one-third of the water systems contained fecal coliforms and/or fecal streptococci. Acinetobacter spp. were detected in 38% of the groundwater supplies at an arithmetic mean density of 8 CFU/100 ml and were present in 16% of the water supplies in the absence of total coliforms, posing some concern about the usefulness of total coliforms as indicators of the presence of this opportunistic pathogen. Slime production, a virulence factor for A. calcoaceticus, was not significantly different between well water isolates and clinical strains, suggesting some degree of pathogenic potential for strains isolated from groundwater. In addition, several Acinetobacter isolates were able to interfere with sheen production by some coliform bacteria on M-Endo medium, adding further to the possible significance of Acinetobacter spp. in groundwater supplies.     

Detection of Acinetobacter spp. in rural drinking water supplies

A bacteriological survey was conducted of untreated, individual groundwater supplies in Preston County, W.Va. Nearly 60% of the water supplies contained total coliforms in excess of the U.S. Environmental Protection Agency maximum contaminant level of 1 CFU/100 ml. Approximately one-third of the water systems contained fecal coliforms and/or fecal streptococci. Acinetobacter spp. were detected in 38% of the groundwater supplies at an arithmetic mean density of 8 CFU/100 ml and were present in 16% of the water supplies in the absence of total coliforms, posing some concern about the usefulness of total coliforms as indicators of the presence of this opportunistic pathogen. Slime production, a virulence factor for A. calcoaceticus, was not significantly different between well water isolates and clinical strains, suggesting some degree of pathogenic potential for strains isolated from groundwater. In addition, several Acinetobacter isolates were able to interfere with sheen production by some coliform bacteria on M-Endo medium, adding further to the possible significance of Acinetobacter spp. in groundwater supplies.     

Magnitude of pollution indicator organisms in rural potable water.

A total of 460 water samples were randomly drawn from the potable water supply sources of rural communities in three counties of South Carolina. About 10% of the population, not incorporated in municipalities, was sampled. The samples were tested for total coliforms, Escherichia coli, and fecal streptococci. Significant levels of these pollution indicator organisms were detected in almost all the water supplies. Total coliforms were the most common, and only 7.5% of the water supplies were uncontaminated. E. coli, considered a reliable indicator of recent and dangerous pollution, was observed in 43% of the water supplies. Statistical analyses indicated that the bacterial populations, especially E. coli, were associated with the supply source depth and its distance from the septic tank. Total coliform counts were also weakly correlated to the pH of the water.     

Human origin of Bacteroides fragilis bacteriophages present in the environment

Bacteroides fragilis HSP40 phages have been detected in waters with various levels of fecal contamination of human origin. The average numbers of B. fragilis phages present in sewage water reached 5.3 x 10(3) per 100 ml of water. We found a number 1,000 times lower in a river contaminated with domestic sewage only, in which the levels of fecal coliforms and fecal streptococci were 10,000 times lower than those found in raw sewage. In addition, B. fragilis phages were not found in significant numbers in slaughterhouse wastewaters. They were not present in fecal-polluted waters containing fecal contamination from wildlife only. Although the number of B. fragilis phages present in contaminated waters was lower than the number of coliphages, their presence indicated human fecal contamination. It is also shown that Bacteroides phages are only able to multiply under anaerobic conditions in the presence of nutrients, and they cannot multiply in natural waters and sediments.     

Bacterial pollution indicators in the intestinal tract of freshwater fish

A study was made of the occurrence, distribution, and persistence of coliforms, fecal coliforms, and fecal streptococci in the intestinal tract of freshwater fish. A total of 132 fish representing 14 different species were used in various phases of these experiments. Examination of the intestinal contents of 78 fish from moderately polluted sections of the Little Miami River indicated that fecal coliform densities were lowest in bluegills (less than 20 per gram) and highest in catfish (1,090,000 per gram). Levels of fecal streptococci for these two species were 220 and 240,000 per gram, respectively. The occurrence of fecal coliforms in fish caught in this stream reflected the warm-blooded-animal-pollution level of the water. All fish used in this phase of the study were caught during July, August, and September when the water temperatures were between 13 and 18 C. The fate of fecal coliforms and Streptococcus faecalis in the fish intestine indicated that these organisms can probably survive and multiply when fish and water temperatures are 20 C or higher, but only when the organisms are retained in the gut for periods beyond 24 hr. Based on the biochemical reactions for 3,877 coliform strains isolated from 132 freshwater fish of 14 different species, 91.4% of all strains were composed of five IMViC types. In a similar study of the biochemical reactions of 850 streptococci isolated from the intestinal tract of 55 freshwater fish, the predominant strains included S. faecalis and various closely associated biotypes. No consistently recurring pattern for either coliforms or streptococci could be developed to identify species of fish investigated. The composition of the intestinal flora is, however, related in varying degree to the level of contamination of water and food in the environment.     

Human origin of Bacteroides fragilis bacteriophages present in the environment.

Bacteroides fragilis HSP40 phages have been detected in waters with various levels of fecal contamination of human origin. The average numbers of B. fragilis phages present in sewage water reached 5.3 x 10(3) per 100 ml of water. We found a number 1,000 times lower in a river contaminated with domestic sewage only, in which the levels of fecal coliforms and fecal streptococci were 10,000 times lower than those found in raw sewage. In addition, B. fragilis phages were not found in significant numbers in slaughterhouse wastewaters. They were not present in fecal-polluted waters containing fecal contamination from wildlife only. Although the number of B. fragilis phages present in contaminated waters was lower than the number of coliphages, their presence indicated human fecal contamination. It is also shown that Bacteroides phages are only able to multiply under anaerobic conditions in the presence of nutrients, and they cannot multiply in natural waters and sediments.     

Incidence of bacterial enteropathogens in foods from Mexico

We examined food consumption patterns of U.S. students temporarily living in Guadalajara, Mexico. Consumption of foods prepared in Mexican homes was associated with an increased risk of acquisition of diarrhea. Foods from commercial sources and private Mexican homes in Guadalajara were subsequently examined for contamination with coliforms, fecal coliforms, and bacterial enteropathogens. For comparison, selected restaurant foods were obtained in Houston, Tex. Food obtained from Mexican homes showed generally higher counts of coliforms and fecal coliforms than those obtained from commercial sources in Mexico and Houston. The foods in Mexico, both from homes and commercial sources, commonly contained Escherichia coli and occasionally enterotoxigenic E. coli. Foods in Houston were not contaminated with E. coli or enterotoxigenic E. coli. Salmonella (17 isolates), Shigella (4 isolates), and Aeromonas hydrophila (1 isolate) were found only in the foods obtained from Mexican homes. Enterotoxigenic non-E. coli Enterobacteriaceae was recovered with approximately equal frequency from all food sources.     

Sperm quality differences between the rainy and dry seasons in captive black-handed spider monkeys (Ateles geoffroyi)

The present work provides an assessment of sperm measures (concentration, motility, viability, etc.) of three black-handed spider monkeys (Ateles geoffroyi) during the rainy and dry seasons in Mexico City, as well as an evaluation of the between-subjects variability of sperm quality. Twenty samples obtained by rectal electroejaculation and digested with trypsin were evaluated. The results showed that during the dry season (n = 9) the semen samples were of better quality than those obtained during the rainy season (n = 11). The individual animals showed differences in sperm concentration, although there were no differences in sperm quality.