Free-living pathogenic and nonpathogenic amoebae in Maryland soils.

Tests for potentially pathogenic amoebae were carried out on soil samples from the following sites: (i) farmlands fertilized with municipal sewage wastes, (ii) a stream receiving sewage effluent from a sludge lagoon, (iii) a ravine receiving storm runoff from a cattle farm, (iv) farmlands not fertilized with sewage wastes, and (v) a vegetated shoreline of a waterfront estate not used for farming or livestock production. Study sites were located on the eastern shore of Maryland, bordered to the north by Delaware and to the south by Virginia. Twenty-four species of soil amoebae, including five potentially pathogenic Acanthamoeba species (members of the family Acanthamoebidae), were identified. All of the sites yielded two or more of the potential pathogens.     

Application of flow cytometry to studies of pathogenic free-living amoebae

Species of small, free-living amoebae of the genera Naegleria and Acanthamoeba can cause fatal amoebic meningoencephalitis. Previous investigations have shown that pathogenic amoebae are associated with thermally altered water. Flow cytometric techniques for identifying species of pathogenic and nonpathogenic amoebae from such water have been developed, using immunofluorescence and fluorescein-bound concanavalin A. Flow cytometry is accomplished with a cytofluorograph, in which cells are dispersed in a suspended carrier liquid and passed in front of a focused argon ion laser beam. Cells are then distinguished by the degree of scattered light (size) or fluorescence. Flow cytometry techniques have proven efficient for environmental samples, as indicated by the identification of pathogenic Naegleria fowleri and nonpathogenic Naegleri gruberi and Acanthamoeba castellanii isolated from the Savannah River Plant in South Carolina. Cytofluorographic analysis of environmental samples has several advantages over the current methods of isolation and classification of free-living amoebae. With this system, it is possible to rapidly identify species and quantitate mixtures of pathogenic amoebae in environmental samples. Cytofluorographic analysis of amoebic isolates reduces the time presently required to screen environmental sites for pathogenic amoebae. The cytofluorograph permits detection and species identification of nonthermophilic Naegleria spp. and Acanthamoeba spp. that could not easily be isolated for species identification by conventional methods. Other advantages of flow cytometry over fluorescent microscopy include a high degree of statistical precision due to the large numbers measured, high immunofluorescent titers, and elimination of subjectivity and fluorescence fading.     

Ecology of cycloheximide-resistant fungi in field soils receiving raw city wastewater or normal irrigation water

The effect of raw city wastewater irrigation on biodiversity and population densities of a cycloheximide-resistant (CH) fungal community was studied in 13 field soils receiving either raw city wastewater or normal irrigation, and in raw city wastewater in the Nablus area, using the hair baiting technique (HBT) and a surface soil dilution plating (SSDP) technique. Three of these fields [one had been receiving raw city wastewater for more than ten years and was designated a heavily polluted field, and the other 2 were cultivated for the first time and were either irrigated with raw city wastewater (newly polluted field) or normal irrigation water (nonpolluted)], were sampled 4–7 times over a 9-month period. The other ten fields, which had been under raw city wastewater irrigation for more than 10 years, were sampled only once. Fifty-seven CH-resistant species belonging to 18 genera were recovered, of which 49 species were recovered from soil habitats and 28 species from raw city wastewater. The HBT had shown to be more efficient in the isolation of pathogenic and potentially pathogenic fungi including dermatophytes. A higher percentage of this group of fungi was recovered from the three main field soils studied using HBT (70% of all isolates), than the SSDP (35.5%); no dermatophytes were recovered by the SSDP method. Two dermatophytes (Microsporum gypseum, and Trichophyton ajelloi), and five more fungi (Arthroderma cuniculi, A. curreyi, Chrysosporium keratinophilum, C. tropicum, and C. pannorum), were recovered from these habitats. Wastewater irrigation seemed to have affected the fungal population densities, with the highest population densities being found in the heavily polluted field soil, while lower population densities were found in the nonpolluted field soil. Increases in organic matter were also observed as a result of sewage effluent irrigation. However, basic similarities in the biodiversity of CH-resistant fungal communities existed in nonpolluted and polluted field soils, and raw city wastewater. Comparable numbers of fungal species were recovered from the three main field soils. The species most commonly found in those habitats included: Alternaria alternata, Aspergillus candidus, Geotrichum candidum, and Paecilomyces lilacinus. Field soils receiving either raw city wastewater or normal irrigation water, were found to be rich in pathogenic and potentially pathogenic CH-resistant fungi, including dermatophytes, with raw city wastewater yielding the highest percentage (81%), followed by the newly wastewater irrigated field (77.7%), the nonpolluted field (67%), and the heavily polluted field (63.4%). Hygienic measures should therefore be taken to control the spread of these fungi in the environment of human communities, and to avoid mycotic infections among farmers. This revised version was published online in August 2006 with corrections to the Cover Date.     

Determinants of community organization of a South African mesic grassland

Abstract. Question: What is the long‐term influence of nutrient availability, productivity and soil pH on grassland community organization? Location: Ukulinga research farm, KwaZulu‐Natal, South Africa. Methods: The influence of fertilization on soil pH, nitrogen (N) and phosphorus (P) on variation in plant traits, community composition and species richness were examined in a 50‐year grassland fertilization experiment. Results: Averaged over 30 years, above‐ground net primary production (ANPP) was 337, 428 and 518 g.m^‐2 in sites not fertilized, fertilized with N, and fertilized with N plus P respectively. ANPP depended directly on N‐fertilization but not on P‐fertilization or liming, and responded positively to the interaction of N (first limiting nutrient) and P (second limiting nutrient). Short narrow‐leaved grass species —Themeda triandra, Tristachya leucothrix and Setaria nigrirostris— dominated sites of lowest ANPP where N was limiting (unfertilized, P‐fertilized or limed sites). A tall narrow‐leaved species, Eragrostis curvula, dominated sites of intermediate ANPP where P was limiting (N‐fertilized sites). By contrast, a tall broad‐leaved species, Panicum maximum, dominated the most productive sites where neither N nor P were limiting (N‐ and P‐fertilized sites). Certain species responded to liming and type of N‐fertilizer apparently because of their effects on soil pH. N‐fertilization reduced the density of herbaceous dicots (forbs) from 14 (unfertilized) to two (high N, no P, no lime) and five species per m² (high N, no P, limed). This effect was attributed to increased ANPP and a decrease in soil pH from 4.6 (KCl) in unfertilized sites to 3.49 (high N, no lime) and 4.65 (high N and lime). Soil acidification had no effect on grass species richness but influenced the abundance of certain species. Conclusions: Grassland community organization is determined not only by the influence of N availability, but also by the hierarchical interaction of N and P availability, in part through their compounded effect on ANPP, and by individualistic species responses to soil pH.     

PCR-based identification of thermotolerant free-living amoebae in Italian hot springs

Several thermal areas, also used for leisure purposes, may represent suitable habitats for free-living amoebae (FLAs), but few studies have been carried out in search for these organisms. The aim of this study was to assess the presence and distribution of FLAs by culture detection and molecular identification, over a one year-round sampling of two sites in Central Italy. Two geothermal springs (Site A and Site B) were investigated for a total of 36 water samples. Four sets of primers were used to amplify FLA DNA from all cultures positive for amoebic growth at both 37 °C and 45 °C. Overall, 33 (91.6%) water samples produced PCR amplification. Eleven taxa were identified. The array of identified species varied over the sampling period, and differed between the two hot springs, Site A harbouring 11 taxa compared to 5 of site B. However, both sites were characterized by the most common species Vermamoeba vermiformis and Naegleria australiensis. Acanthamoeba genotypes T4 and T15 were found at low frequency. Differences in the composition between the two sites could reflect environmental changes in biotic and chemical/physical parameters. From a public health perspective, the detection of potentially pathogenic amoebae could unveil a potential risk for humans.     

25S rDNA-based molecular monitoring of glomalean fungi in sewage sludge-treated field plots

Recycling of sewage wastes in agriculture is likely to affect the biological activity of soils through contamination of ecosystems by pathogens and metallic or organic micropollutants. The impact of sewage sludge spreading under field conditions on arbuscular mycorrhiza (AM) formation by a community of glomalean fungi was evaluated using a nested polymerase chain reaction (PCR) and discriminating primers based on 25S rDNA polymorphisms to detect different fungal species within root systems. Medicago truncatula was grown in soil of field plots amended or not with a composted sewage sludge, spiked or not with organic or metallic micropollutants. Overall AM development in roots decreased with sewage sludge application, and the relative abundance of five AM fungal morphotypes in root fragments was modified by the input of composted sludges. Sewage sludge spiked or not with organic pollutants had a generally positive effect on the relative diversity of AM fungal populations in planta , whereas after spreading of the sludge spiked with metallic pollutants, no variation was observed in the abundance of different species.     

Ecological risk assessment of toxic elements,microbial loads,parasites and antinutritional factors in Telfairia occidentalis grown on sewage contaminated soil

The aim of this study was to evaluate the ecological health risk assessment of toxic elements, microbial load, and anti-nutrient constituent build-up in vegetables, Talfairia occidentalis, grown on sewage contaminated soil. Samples of soil and vegetables were taken from farms near the dumpsites (sites A). As a control, samples were taken from an area with no dump sites (sites B). Atomic Absorption Spectrophotometry was used to determine the potentially toxic elements. The basic titrimetric method was used to estimate Phytate and Oxalate in the vegetables in order to assess anti-nutrient constituents. The content of cyanogenic glycosides, tannins, and alkaloids in vegetable samples was determine using a standard method. Total heterotrophic bacteria, E. coli, total coliform, faecal coliform, Staphylococcus aureus, Salmonella, and intestinal parasites were all determined using APHA standard methods. For all samples analyzed, the concentrations of metals – Cr, Zn, Pb, Cu, and Ni in site A were higher than those in site B. The soil and vegetable samples differed from the controls by a significant amount (P <0.05). Vegetable samples from site A were mainly infected with faecal coliform TO (9.7 × 10⁵cfu/g) and other bacteria at levels higher than the recommended levels. In contrast to samples from site B, which were not infected with human intestinal helminth parasites, Ascaris lumbricoides ova and Entamoeba histolytica cryst were present in vegetables in site A samples. The majority of the values obtained for the sewage dumpsite were significantly higher than the values recorded by the World Health Organization (WHO). However, sewage wastes, anthropogenic sources, and atmospheric depositions cause higher concentrations of heavy metals, anti-nutrients, and microbial load around dumpsites, which bioaccumulate in vegetables through uptake from the soil and eventual entry into the food chain. The overall evaluation found that consumption of vegetables grown on sewage soil poses a risk of heavy metals, microbial loads, and anti-nutrients adversely affecting human health.     

Effects of elevated nitrogen and exotic plant invasion on soil seed bank composition in Joshua Tree National Park

We investigated the effects of exotic species invasion and 3?years of nitrogen (N) fertilization on the soil seed bank in Joshua Tree National Park, California, USA at four sites along an N deposition gradient. We compared seed bank composition and density in control (no N added) and fertilized (30?kg?N?ha^?1?year^?1) plots to determine if the seed bank would reflect aboveground changes due to N fertilization. Soil samples were collected and germinated in a greenhouse over 2?years. In the field, invasive species cover responded positively to N fertilization. However, we did not observe increased seed density of exotic invasive species in fertilized plots. While no significant differences were detected between treatments within sites, exotic invasive grass seeds overwhelmed the seed bank at all sites. Significant differences between sites were found, which may be due to differences in level of invasion, historic N deposition, and soil surface roughness. Sites experiencing low N deposition had the highest seed bank species richness for both control and fertilized treatments. Aboveground plant density did not correlate well with seed bank density, possibly due to the inherent patchiness of soil seed banks and differential ability of species to form seed banks. This seed bank study provided insight into site-specific impacts on native versus invasive species composition of soil seed banks, as well as magnitude of invasion and restoration potential at invaded sites.     

Biological testing of a digested sewage sludge and derived composts

Aiming to evaluate a possible loss of soil habitat function after amendment with organic wastes, a digested sewage sludge and derived composts produced with green residues, where biologically tested in the laboratory using soil animals (Eisenia andrei and Folsomia candida) and plants (Brassica rapa and Avena sativa). Each waste was tested mimicking a field application of 6ton/ha or 12ton/ha. Avoidance tests did not reveal any impact of sludge and composts to soil biota. Germination and growth tests showed that application of composts were beneficial for both plants. Composts did not affect earthworm's mass increase or reproduction, but the highest sludge amendment revealed negative effects on both parameters. Only the amendment of composts at the highest dose originated an impairment of springtails reproductive output. We suggest that bioassays using different test species may be an additional tool to evaluate effects of amendment of organic wastes in soil. Biological tests are sensitive to pollutants at low concentrations and to interactions undetected by routine chemical analysis.     

Potentially human pathogenic Acanthamoeba isolated from a heated indoor swimming pool in Switzerland

Some free-living amoebae, including some species of the genus Acanthamoeba, can cause infections in humans and animals. These organisms are known to cause granulomatous amebic encephalitis (GAE) in predominantly immune-deficient persons. In the present study, we isolated a potentially human pathogenic Acanthamoeba isolate originating from a public heated indoor swimming pool in Switzerland. The amoebae, thermophilically preselected by culture at 37 °C, subsequently displayed a high thermotolerance, being able to grow at 42 °C, and a marked cytotoxicity, based on a co-culture system using the murine cell line L929. Intranasal infection of Rag2-immunodeficient mice resulted in the death of all animals within 24 days. Histopathology of brains and lungs revealed marked tissue necrosis and hemorrhagic lesions going along with massive proliferation of amoebae. PCR and sequence analysis, based on 18S rDNA, identified the agent as Acanthamoeba lenticulata. In summary, the present study reports on an Acanthamoeba isolate from a heated swimming pool suggestive of being potentially pathogenic to immunocompromised persons.     

Testate amoebae taxonomy and trait diversity are coupled along an openness and wetness gradient in pine-dominated Baltic bogs

Sphagnum peatlands host a high abundance of protists, especially testate amoebae. Here, we designed a study to investigate the functional diversity of testate amoebae in relation to wetness and forest cover in Baltic bogs. We provided new data on the influence of openness/wetness gradient on testate amoebae communities, showing significant differences in selected testate amoebae (TA) traits. Three key messages emerged from our investigations: 1) we recorded an effect of peatland surface openness on testate amoebae functional traits that led us to accept the hypothesis that TA traits differ according to light intensity and hydrology. Mixotrophic species were recorded in high relative abundance in open plots, whereas they were nearly absent in forested sites; 2) we revealed a hydrological threshold for the occurrence of mixotrophic testate amoebae that might be very important in terms of peatland functioning and carbon sink vs. source context; and 3) mixotrophic species with organic tests were nearly absent in forested sites that were dominated by heterotrophic species with agglutinated or idiosomic tests. An important message from this study is that taxonomy of TA rather indicates the hydrological gradient whereas traits of mixotrophs the openness gradient.     

Impact of Fertilizing with Raw or Anaerobically Digested Sewage Sludge on the Abundance of Antibiotic-Resistant Coliforms,Antibiotic Resistance Genes,and Pathogenic Bacteria in Soil and on Vegetables at Harvest

The consumption of crops fertilized with human waste represents a potential route of exposure to antibiotic-resistant fecal bacteria. The present study evaluated the abundance of bacteria and antibiotic resistance genes by using both culture-dependent and molecular methods. Various vegetables (lettuce, carrots, radish, and tomatoes) were sown into field plots fertilized inorganically or with class B biosolids or untreated municipal sewage sludge and harvested when of marketable quality. Analysis of viable pathogenic bacteria or antibiotic-resistant coliform bacteria by plate counts did not reveal significant treatment effects of fertilization with class B biosolids or untreated sewage sludge on the vegetables. Numerous targeted genes associated with antibiotic resistance and mobile genetic elements were detected by PCR in soil and on vegetables at harvest from plots that received no organic amendment. However, in the season of application, vegetables harvested from plots treated with either material carried gene targets not detected in the absence of amendment. Several gene targets evaluated by using quantitative PCR (qPCR) were considerably more abundant on vegetables harvested from sewage sludge-treated plots than on vegetables from control plots in the season of application, whereas vegetables harvested the following year revealed no treatment effect. Overall, the results of the present study suggest that producing vegetable crops in ground fertilized with human waste without appropriate delay or pretreatment will result in an additional burden of antibiotic resistance genes on harvested crops. Managing human exposure to antibiotic resistance genes carried in human waste must be undertaken through judicious agricultural practice.     

The pathogenic amoeboflagellate Naegleria fowleri: environmental isolations, competitors, ecologic interactions, and the flagellate-empty habitat hypothesis

From several surveys of environmental sites, the virulent human pathogen, Naegleria fowleri, was isolated from a pond in Georgia, a sewage treatment plant in Missouri, and from the Potomac and Anacostia rivers near and in Washington, D.C. Widely scattered, sparse populations seemed only a potential threat to human health at the time of sampling. The data support an estimate that the sites sampled contain 10,000 typical, low temperature, bactivorous amoebae for each heat tolerant amoeba able to grow at 45 degrees C. Heat tolerant competitors were much more common than N. fowleri. Naegleria lovaniensis, which is heat tolerant but nonpathogenic, was isolated from and downstream from an open air thermal pollution temperature gradient. Hot piles of composting sewage sludge yielded no amoeboflagellates, many heat tolerant (45-49 degrees C) amoebae, and one thermophilic (52 degrees C) Acanthamoeba. Features of the methods used include two-stage incubation to increase isolation of sparse organisms and distinction of N. fowleri from almost all other amoebae on agar plates. The flagellate-empty habitat hypothesis postulates a general model in which human intervention and/or natural events remove usual competitors and the ability to transform to a motile flagellate confers an advantage in recolonizing.     

Are there any agricultural effects on the capture rates of male euglossine bees (Apidae: Euglossini)?

During 30 days male euglossine bees were bait-sampled at 12 sites, in the central Pacific coast of Colombia (ten days and four sites at each of three adjacent habitats: farmlands, highly disturbed forest and less disturbed forest) and 487 individuals were captured. Most captured individuals belonged to six species, five widely distributed through the American tropics and an endemic species. Two of the frequently captured species presented no different abundances between habitats, while the other four (67.97% of all the captured individuals), all of them big sized or long-tongued, were more frequently captured at the farmlands. A cluster analysis of the data matrix for the 23 captured species and the 12 sampling sites, grouped together the farmland sites, apart from the forest sites. It is proposed that male euglossine bees from generalist, long-tongued or big sized species, forage frequently at the farmlands, where fragrance or nectar resources may be clumped, less diverse, and present an access restricted by deep corollas or by microclimatic conditions of high temperature and low humidity.     

Epizootiology of Eustrongylides ignotus in Florida: distribution,density, and natural infections in intermediate hosts

A total of 63,451 fish, representing 39 species, was collected from 176 foraging sites used by ciconiiform wading birds in peninsular Florida (USA) and examined for larvae of Eustrongylides ignotus. Infected fish were identified from 30 (17%) of the sites, all of which had been altered by human disturbance such as removal of sediment to construct ditches and dikes, improve water flow, or increase storage capacity and had a history of receiving anthropogenic nutrients such as sewage effluent, urban runoff, or agricultural runoff. The mosquitofish (Gambusia holbrooki) and several species of sunfish (Centrarchidae) were the most important intermediate hosts. Infected fish were not collected at any of the unaltered sites. A total of 10,508 oligochaetes (representing 36 species) was identified from 22 sites that had fish infected with E. ignotus and 36 sites where no infected fish were collected. None of the oligochaetes was infected with larvae of E. ignotus. Immature tubificids without hair setae (probably Limnodrilus sp.), Dero digitata, and L. hoffmeisteri were the most abundant oligochaetes at sites where infected fish occurred, making up 78% of the total collected. Compared to unaltered sites, altered sites were characterized by higher mean densities of fish and oligochaetes; surface waters with decreased dissolved oxygen and increased total nitrogen, total phosphorus, and chlorophyll-a; sediments with higher soil oxygen demand and total phosphorus; larger grain sizes; and higher percentage emergent vegetation and grasses.     

Is the relation between nutrient supply and biodiversity co‐determined by the type of nutrient limitation?

Correlative studies have shown a ‘hump‐backed’ relation between the vegetation N:P ratio and plant species diversity with the highest diversity at balanced N:P ratios (between 10 and 14). We tested the hypothesis that adding growth‐limiting nutrients to mesotrophic grasslands that were in shortage of either N (N:P ratio<10) or P (N:P ratio>14) would lead to an increase of plant diversity. Thereto, we studied the effects of long‐term (11 yr) experimentally increased N and/or P supply on soil nutrient pools, vegetation nutrient dynamics and biodiversity in a riverine grassland in the Netherlands with a low soil N:P ratio (N shortage) and a peat grassland with a high soil N:P ratio (P shortage), respectively. Eleven years of nutrient addition hardly had any effects on the total stocks of C, N and P in the soils of both sites, due to the large size of the soil nutrient pools already present and to the management at both sites (annual hay‐making and ‐removal). However, in the riverine grassland the treatments increased the cycling of the small pool of labile N and P compounds resulting in large increases in annual fluxes of especially N. In the unfertilised controls, species establishments balanced more or less species losses during an 11 year period, thus leading to a dynamic equilibrium of the species pool. However, contrary to our hypothesis, addition of the growth‐limiting nutrient led at both sites to a reduction of species diversity even when total biomass remained below critical levels. Species diversity and species evenness were strongly determined by N mineralisation and to a lesser extent by total soil N and extractable P, respectively. Total aboveground biomass of the vegetation was determined by total soil N. Our study shows that patterns found in correlative studies of the relation between plant diversity and soil and vegetation N:P ratio can not be translated into successful experimental manipulations to enhance biodiversity. The most likely explanation is that colonization limitation occurred in the fertilized plots and that not sufficient diaspores of potentially new species could reach and/or colonize the plots to compensate for the species extinctions as a result of increased nutrient supply.     

Cocultivation of the amoeba Naegleria fowleri and the amoebicin- producing strain Bacillus licheniformis M-4.

Antagonism between Bacillus licheniformis M-4 and the pathogenic amoeba Naegleria fowleri HB-1 during cocultivation was influenced by the composition of the medium and the initial amoeba/bacterium ratio. While a ratio of 50 caused complete lysis of amoebae in soil extract with 0.3% glucose (SEG) before 72 h, this ratio had to be at least 12-fold lower in order to obtain similar results in Cline medium. Sporulation of B. licheniformis M-4 took place much earlier in SEG. Amoebicin production was stimulated by the presence of amoebae by either shortening the time of production (as in SEG) or increasing the amount of amoebicins released (as in Cline medium). Electron microscopy showed that amoebae cocultivated in the Cline medium contained bacteria enclosed in digestive vacuoles, while amoebae from SEG cocultures did not.     

Persistence of poliovirus 1 in soil and on vegetables grown in soil previously flooded with inoculated sewage sludge or effluent.

Land disposal of sewage sludge and effluent is becoming a common practice in the United States. The fertilizer content and humus value of such wastes are useful for agricultural purposes, and the recycling of sewage onto the land eliminates many of our stream pollution problems. The potential exists for crops grown in such irrigated soil to be contaminated by viruses that may be present in the sewage. Studies were initiated to determine viral persistence in soil and on crops grown under natural conditions in field plots that had been flooded to a depth of 1 inch (2.54 cm) with poliovirus 1-inoculated sewage wastes. Lettuce and radishes were planted in sludge- or effluent-flooded soil. In one study, the vegetables were planted 1 day before flooding, and in another they were planted 3 days after the plots were flooded. Survival of poliovirus 1 in soil irrigated with inoculated sewage sludge and effluent was determined during two summer growing seasons and one winter period. The longest period of survival was during the winter, when virus was detected after 96 days. During the summer, the longest survival period was 11 days. Poliovirus 1 was recovered from the mature vegetables 23 days after flooding of the plots had ceased. Lettuce and radishes are usually harvested 3 to 4 weeks after planting.     

Persistence of poliovirus 1 in soil and on vegetables grown in soil previously flooded with inoculated sewage sludge or effluent.

Land disposal of sewage sludge and effluent is becoming a common practice in the United States. The fertilizer content and humus value of such wastes are useful for agricultural purposes, and the recycling of sewage onto the land eliminates many of our stream pollution problems. The potential exists for crops grown in such irrigated soil to be contaminated by viruses that may be present in the sewage. Studies were initiated to determine viral persistence in soil and on crops grown under natural conditions in field plots that had been flooded to a depth of 1 inch (2.54 cm) with poliovirus 1-inoculated sewage wastes. Lettuce and radishes were planted in sludge- or effluent-flooded soil. In one study, the vegetables were planted 1 day before flooding, and in another they were planted 3 days after the plots were flooded. Survival of poliovirus 1 in soil irrigated with inoculated sewage sludge and effluent was determined during two summer growing seasons and one winter period. The longest period of survival was during the winter, when virus was detected after 96 days. During the summer, the longest survival period was 11 days. Poliovirus 1 was recovered from the mature vegetables 23 days after flooding of the plots had ceased. Lettuce and radishes are usually harvested 3 to 4 weeks after planting.     

Effect of chlorination on antibiotic resistance profiles of sewage-related bacteria

A total of 1,900 lactose-fermenting bacteria were isolated from raw sewage influent and chlorinated sewage effluent from a sewage treatment plant, as well as from chlorinated and neutralized dilute sewage, before and after a 24-h regrowth period in the laboratory. Of these isolates, 84% were resistant to one or more antibiotics. Chlorination of influent resulted in an increase in the proportion of bacteria resistant to ampicillin and cephalothin, the increase being most marked after regrowth occurred following chlorination. Of the other nine antibiotics tested, chlorination resulted in an increased proportion of bacteria resistant to some, but a decrease in the proportion resistant to the remainder. Multiple resistance was found for up to nine antibiotics, especially in regrowth populations. Identification of about 5% of the isolates showed that the highest proportion of Escherichia coli fell in untreated sewage. Some rare and potentially pathogenic species were isolated from chlorinated and regrowth samples, including Yersinia enterocolitica, Yersinia pestis, Pasteurella multocida, and Hafnia alvei. Our results indicate that chlorination, while initially lowering the total number of bacteria in sewage, may substantially increase the proportions of antibiotic-resistant, potentially pathogenic organisms.