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The incidence and level of contamination of British fresh sausages and ingredients with salmonellas.

A five-tube most probable number (MPN) method, with the pre-enrichment and enrichment stages, was used in a study of the incidence of salmonella contamination of British fresh sausages and the ingredients used in their manufacture. All samples were taken from a large factory in the course of routine production. There was an incidence of 65% contamination of pork (n = 20) and 55% (n = 20) in pork and beef sausages. The incidences of contamination of uncooked ingredients varied from 95% for mechanically recovered meat (n = 20) to 10% for another type of meat. Cooked and/or dried ingredients were rarely contaminated with these organisms, and when contamination occurred, coliforms were also important. The numbers of salmonellas isolated ranged from 7-40 for pork sausages, from 8-24 for beef and pork sausages and from 0.8-378 organisms/g for ingredients. The following salmonella serotypes were isolated (ranked in descending order or incidence): S. derby, S. dublin, S. newport, S. stanley. S. typhimurium, S. heidelberg, S. infantis and S. agona.

Animals↗

Experimental studies on environmental contamination with infected blood during haemodialysis.

To assess the relative importance of different postulated modes of spread of hepatitis B in dialysis units, blood charged with various tracer organisms was used in simulated haemodialysis runs in four laboratories, and the resulting contamination of equipment and environment was measured semi-quantitatively. Some airborne spread of the tracer organism occurred when tubing containing contaminated blood was needled as the "patient" went on and came off the dialyser. Virtually no small airborne particles could be demonstrated however in simulated emergencies in which a blood line was disconnected, or even when bottles of blood were dropped on to a hard floor from a height of 2 metres. Bacillus globigii spores from contaminated blood leaked in small numbers into the dialysing fluid through apparently intact coils. T3 phage, with a particle size of the same order as hepatitis B virus, passed in small quantities through the membrane of a Kiil dialyser from blood to dialysing fluid and also in the reverse direction when added to the header tank. A number of other dialysers were also permeable to phage. Visual assessment of the appropriate moment for inserting the venous line into the "patient" at the onset of dialysis was shown to be unreliable, as the displaced fluid from the end of the venous line was already contaminated before it contained visible red blood cells. Considerable contamination of exposed surfaces and of the buttons on the proportionating unit cabinet occurred. Minor visible splashing of blood was a common-place of the laboratory experiments and was shown to be also a common event during routine haemodialysis in two of the dialysis units taking part in the studies.

Air Microbiology↗

Observations on environmental contamination in a microbiological laboratory.

Contamination of a laboratory environment with pathogenic or non-pathogenic micro-organisms may be relevant to safety of technicians and quality of technical performance. Two widely separated incidents in 1968 and 1974 initiated a study of aspects of the laboratory environment. Water-baths, water of syneresis and portions of salmonella cultures spurting out of the sterilizing flame were examined. The water of water-baths was shown to be contaminated from the fluid cultures incubated in them. This raised questions of potential cross-contamination and reporting of false positives. Water of syneresis was sometimes contaminated with salmonellas. A few quantitative counts were made. The range of counts varied between 16 salmonellas per ml. and 13,000,000 salmonellas per ml. Five hundred portions of salmonella cultures and 571 portions of Shigella sonnei cultures which had spurted from the sterilizing flame were examined. All these samples failed to grow salmonellas or shigellas. Precautions necessary to avoid environmental contamination are briefly discussed.

Bacteria↗

Pseudobacteremia traced to cross-contamination by an automated blood culture analyzer.

Twenty-three episodes of pseudobacteremia occurring over a seven-month period were traced to cross-contamination by the automated blood culture analyzer (BACTEC 460) used in the microbiology laboratory. An epidemiologic investigation of an unusual cluster of three patients with pseudobacteremia caused by oxacillin-resistant Staphylococcus aureus led to the identification of the problem. Mock trials of the blood culture procedure confirmed that the blood culture analyzer was the source of contamination. After the needle sterilizer was replaced by the manufacturer, the problem of cross-contamination abated. Contamination of sterile blood cultures by an instrument intended to identify bacteremia rapidly may lead to incorrect diagnosis, unnecessary administration of antibiotics and prolonged hospitalization. Because of the widespread use of automated blood culture analyzers in the US, physicians, microbiologists, and infection control personnel should be alert to the possibility of cross-contamination and the subtle way in which it may present.

Blood↗

Determinants of Legionella pneumophila contamination of water distribution systems: 15-hospital prospective study.

We conducted a prospective environmental study for Legionella pneumophila in 15 hospitals in Pennsylvania. Hot water tanks, cold water sites, faucets, and showerheads were surveyed four times over a one-year period. Sixty percent (9/15) of hospitals surveyed were contaminated with L pneumophila. Although contamination could not be linked to a specific municipal water supplier, most of the contaminated supplies came from rivers. Parameters found to be significantly associated with contamination included elevated hot water temperature, vertical configuration of the hot water tank, older tanks, and elevated calcium and magnesium concentrations of the water (P less than 0.05). This study suggests that L pneumophila contamination could be predicted based on design of the distribution system, as well as physicochemical characteristics of the water.

Cross Infection↗

Faecal contamination of water and fingertip-rinses as a method for evaluating the effect of low-cost water supply and sanitation activities on faeco-oral disease transmission. I. A case study in rural north-east Thailand.

Most villagers in north-east Thailand carry water to their homes and store it in separate containers depending on its subsequent use. In one village, information on water use was collated with the bacteriological quality of stored water, water sources and fingertip-rinses. Stored water quality was a function of water-related activities rather than quality at source (P less than 0.0001). Specifically water used for toilet, washing dishes and cooking-related activities was much more contaminated with faecal bacteria than that used for drinking and cooking. Salmonella spp. was significantly more common in water used for washing dishes than drinking (P less than 0.05). Escherichia coli contamination of fingertip-rinses was strongly associated with the individual's activity prior to testing (P less than 0.0001); child care, food and water-related activities produced much higher levels of fingertip contamination than others. Dirty utensils used for cooking and eating were usually left to soak and faecal bacterial growth occurred in this grossly contaminated soak-water. Cross-contamination via water handling was the main mechanism of stored water pollution. These results were used to develop a hygiene intervention study presented in a companion paper.

Bacteria↗

Occurrence of multiple antibiotic resistance and R-plasmids in gram-negative bacteria isolated from faecally contaminated fresh-water streams in Hong Kong.

The bacterial populations of six freshwater streams in populated areas of the Hong Kong New Territories were studied. There is considerable faecal contamination of these streams, with coliform counts as high as 10(5) c.f.u./ml and the contaminating organisms show a high prevalence of antibiotic resistance and multiple resistance. With direct plating of water samples onto antibiotic-containing media, an average of 49% of the gram-negative bacteria were ampicillin-resistant, 3% chloramphenicol-resistant and 1% gentamicin-resistant. At individual sites resistance to these drugs was as high as 98%, 8% and 3% respectively. More than 70% of strains were resistant to two or more antibiotics, 29% to five or more and 2% to eight or more. A total of 98 patterns of antibiotic resistance were detected with no one pattern predominating. Twenty-eight gram-negative bacterial species were identified as stream contaminants. Escherichia coli was the commonest bacterial species isolated and other frequent isolates were Enterobacter sp., Klebsiella sp. and Citrobacter sp., but no enteric pathogens were detected. The greatest prevalence of resistance and multiple resistance was associated with the heaviest contamination by E. coli. Analysis of selected stream isolates revealed multiple plasmid bands arranged in many different patterns, but multiple antibiotic resistances were shown to be commonly mediated by single transferable plasmids. Faecally-contaminated freshwater streams in Hong Kong may be reservoirs of antibiotic resistance plasmids for clinically-important bacteria.

Animals↗

Routes for salmonella contamination of poultry meat: epidemiological study from hatchery to slaughterhouse.

Data were collected on the prevalence of salmonella at different stages during the life cycle of 18 broiler flocks on different farms as well as during slaughter in different poultry slaughterhouses. For the isolation of salmonella, the highest sensitivity (93.9%) was obtained by enrichment in the semi-solid agar Diasalm. The 'overshoe method' utilizing several pairs of overshoes provided the highest sensitivity for determining the salmonella status of the broilers during rearing. A clear decrease of the relative importance of the first production stages was demonstrated for the salmonella contamination of the end product, whereas horizontal transmission of salmonella to broilers during rearing and to broiler carcasses in the slaughterhouse was shown to be the main determinative factor. Ten of the 18 flocks received a salmonella positive status with the highest shedding occurring during the first 2 weeks of rearing. The shedding of the animals was significantly negatively influenced by the use of subtherapeutic or therapeutic doses of antibiotics. The intake of portable material in the broiler house was identified as the most important risk factor for horizontal transmission. Significant associations were found between the contamination level of a flock and hygiene of the broiler house, feed and water in the broiler house and both animal and non-animal material sampled in the environment. No correlation was found between contamination during the rearing period and contamination found after slaughtering. The presence of faecal material in the transport crates and predominantly the identity of the slaughterhouse seemed to be the determining factors for carcass quality. Improved hygiene management during transport of broilers and in some slaughterhouses could significantly reduce the risk of salmonella contamination of poultry meat.

Animal Husbandry↗

Purification of contaminated peptides and proteins on synthetic membrane surfaces for matrix-assisted laser desorption/ionization mass spectrometry.

Effective MALDI-TOF MS of salt-, detergent-, and glycerol-contaminated peptide and protein samples is accomplished by loading samples onto synthetic membranes, washing away contaminants, adding matrix, and desorbing samples directly from the membrane surface. The method easily removes contaminants which prevent effective MALDI of peptides and proteins from stainless steel surfaces, obviating the need for laborious further purification and associated sample losses. Polyethylene, polypropylene, C8, and C18 surfaces all proved effective at removing contaminants. Scanning electron microscope images of sample surfaces reveal that significantly smaller matrix crystals form on polyethylene and polypropylene surfaces than on stainless steel, C8, or C18 surfaces. Desorption from polyethylene and polypropylene surfaces generates consistently reproducible spectra with better mass resolution than observed for samples desorbed from stainless steel. Improved resolution, combined with reduced intensity of product ion spectra, indicate that peptide and protein molecular ions have less internal energy when desorbed from polyethylene and polypropylene surfaces than from stainless steel. MALDI of contaminated samples can be accomplished by straightforward, on-probe purification, resulting in higher resolution spectra than observed in samples desorbed from stainless steel.

Membranes, Artificial↗

Effects of gasoline formulation on methyl tert-butyl ether (MTBE) contamination in private wells near gasoline stations.

UNLABELLED: The extent of methyl tert-butyl ether (MTBE) contamination in private wells near gasoline stations, which lack the relative protection afforded many public waters supplies through zoning and routine testing, was examined. Samples were collected from 74 private wells near 21 randomly selected gasoline stations and from 21 control wells, one per facility. Two hypotheses are tested: (1) private wells downgradient and close (<0.5 mi) to gasoline stations (case wells) are more likely to have MTBE contamination than private wells upgradient and distant (>1.5 mi) (control wells); and (2) private wells near gasoline stations selling oxygenated gasoline are more likely to have MTBE contamination than private wells near gasoline stations selling conventional gasoline. Data on the concurrence of MTBE and other gasoline constituents are presented. RESULTS: MTBE concentrations ranged from <1.0 micro/L (microg/L) to 61 microg/L, with a mean of 12.0 microg/L. MTBE contamination of > or =1 microg/L was detected more frequently in case wells (28%) than control wells (5%) (p = 0.015). MTBE contamination of > or =1 microg/L occurred more frequently in private wells near facilities selling oxygenated gasoline (38%) than private wells near facilities selling conventional gasoline (20%) (p = 0.11). Statistical significance may have been achieved with a larger sample size. Benzene (0.5 microg/L) was found concurrently with MTBE in only one well, which also had the highest concentration of MTBE.

Benzene↗

Comparative analysis of PAH:DNA adducts formed in lung of mice exposed to neat coal tar and soils contaminated with coal tar.

7H-Benzo[c]fluorene (benzo[c]fluorene) is a major DNA adduct forming component of coal tar in lung of mice. The present study evaluated the types of PAH:DNA adducts formed from different neat coal tar samples and soils contaminated with coal tar. Mice were fed diets containing coal tar either neat or as a contaminant in an environmental soil sample for 14 days, and the types of chemical:DNA adducts formed in lung were evaluated using 32P-postlabeling and HPLC analysis. Three major DNA adducts derived respectively from benzo[b]fluoranthene (B[b]F), benzo[a]pyrene (B[a]P), and benzo[c]fluorene were detected in three of the four neat coal tar samples evaluated. In contrast, only a single major DNA adduct derived from benzo[c]fluorene was observed with the remaining tar sample. Ingestion of coal tar contaminated soil resulted in DNA adducts primarily derived from benzo[c]fluorene and B[b]F; a B[a]P derived DNA adduct was not detected. The DNA adducts derived from benzo[c]fluorene and B[b]F but not B[a]P were also observed with animals fed methylene chloride extracts of three of these soils but not the one designated A1000H soil. However, the extract of A1000H resulted in a B[a]P:DNA adduct being detected along with adducts formed from B[b]F and benzo[c]fluorene. The selective formation of the benzo[c]fluorene:DNA adduct with coal tar contaminated soils indicates that the in vivo systemic bioavailability and/or metabolism of benzo[c]fluorene is relatively high when compared to other DNA adducting hydrocarbons within coal tar. Benzo[c]fluorene may play a critical role in the potential of contaminated soil to induce a toxicological response in animals.

Administration, Oral↗

Ecotoxicity of zinc in spiked artificial soils versus contaminated field soils.

Metal bioavailability is influenced by soil characteristics and aging period. In the present study, both factors were modeled by comparing metal bioavailability in spiked artificial soil and historically contaminated field soils. The chronic toxicity of zinc to Folsomia candida in spiked artificial soils could be predicted with a model based on pH, cation exchange capacity, and total zinc concentration. However, this model could not adequately predict chronic zinc toxicity in contaminated field soils. Porewater concentration and water- and calcium chloride-extracted zinc fractions of the contaminated field soils were lower than those predicted using models developed for spiked artificial soils, indicating that the effect of aging on metal bioavailability should be taken into account. The reproduction of F. candida in contaminated field soils was lower than predicted with models developed using zinc concentration in the porewater and the water- and calcium chloride-extracted fractions in spiked artificial soils. This suggests that these fractions are not the only bioavailable zinc fractions and that dietary metal exposure might also be an important route of uptake under environmentally relevant conditions. Aging and dietary uptake should be studied urgently in order to be able to perform effect-based risk assessments of metal contaminated soils.

Animals↗

Comparison of five in vitro digestion models to study the bioaccessibility of soil contaminants.

Soil ingestion can be a major exposure route for humans to many immobile soil contaminants. Exposure to soil contaminants can be overestimated if oral bioavailability is not taken into account. Several in vitro digestion models simulating the human gastrointestinal tract have been developed to assess mobilization of contaminants from soil during digestion, i.e., bioaccessibility. Bioaccessibility is a crucial step in controlling the oral bioavailability for soil contaminants. To what extent in vitro determination of bioaccessibility is method dependent has, until now, not been studied. This paper describes a multi-laboratory comparison and evaluation of five in vitro digestion models. Their experimental design and the results of a round robin evaluation of three soils, each contaminated with arsenic, cadmium, and lead, are presented and discussed. A wide range of bioaccessibility values were found for the three soils: for As 6-95%, 1-19%, and 10-59%; for Cd 7-92%, 5-92%, and 6-99%; and for Pb 4-91%, 1-56%, and 3-90%. Bioaccessibility in many cases is less than 50%, indicating that a reduction of bioavailability can have implications for health risk assessment. Although the experimental designs of the different digestion systems are distinct, the main differences in test results of bioaccessibility can be explained on the basis of the applied gastric pH. High values are typically observed for a simple gastric method, which measures bioaccessibility in the gastric compartment at low pHs of 1.5. Other methods that also apply a low gastric pH, and include intestinal conditions, produce lower bioaccessibility values. The lowest bioaccessibility values are observed for a gastrointestinal method which employs a high gastric pH of 4.0.

Biological Availability↗

Field evidence for a protistan role in an organically-contaminated aquifer.

The association between protists, bacteria, and dissolved organic carbon (DOC) in an oxygen-depleted, 6 km-long wastewater contaminant plume within a sandy aquifer (Cape Cod, MA) was investigated by comparing abundance patterns along longitudinal and vertical transects and at a control site. Strong linear correlations were observed between unattached bacterial abundance and DOC for much of the upgradient-half of the plume (0.1-2.5 km downgradient from the source) that is characterized by quasi-steady state chemistry. However, a logarithmic decrease was observed between the number of protists supported per mg of DOC and the estimated age of the DOC within the plume. The relatively labile dissolved organic contaminants that characterize the groundwater sampled from the plume < or = 0.1 km downgradient from the contaminant source appeared to indirectly support 3-4 times as many protists (per mg of DOC) as the older, more recalcitrant DOC in the alkylbenzene sulfonate (ABS)-contaminated zone at 3 km downgradient (approximately 30 years travel time). Substantive numbers of protists (>10(4)/cm3) were recovered from suboxic zones of the plume. The higher than expected ratios of protists to unattached bacteria (10 to 100:1) observed in much of the plume suggest that protists may be grazing upon both surface-associated and unattached bacterial communities to meet their nutritional requirements. In closed bottle incubation experiments, the presence of protists caused an increase in bacterial growth rate, which became more apparent at higher amendments of labile DOC (3-20 mgC/L). The presence of protists resulted in an increase in the apparent substrate saturation level for the unattached bacterial community, suggesting an important role for protists in the fate of more-labile aquifer organic contaminants.

Animals↗

Reduction of crop contamination by soil resuspension within the 30-km zone of the Chernobyl nuclear power plant.

A field experiment was conducted within the 30-km zone of the Chernobyl Nuclear Power Plant to analyze whether the application of mulching reduced resuspension of 137Cs contaminated soil in oat (Avena sativa) crops. In 1993, we applied a mulch treatment at a dose of 200 g m(-2), and soil resuspension was measured by estimating soil loadings onto plant surfaces from Ti concentrations in plants. In 1994, two mulch doses were applied, 200 and 50 g m(-2), and we estimated the contribution of soil resuspension by using artificial resuspension collection devices (ARC). In the 1993 experiment between 4.6 and 34.4% of the plant's total 137Cs contamination was attributed to external soil contamination. The mean amount of soil-derived 137Cs attached to vegetation was 124.7 Bq kg(-1)(plant) in control plots and 53.7 Bq kg(-1)(plant) in mulched plots. In the 1994 experiment, covering the soil with a mulch layer decreased the radiocesium content in ARC by about 70%. Results obtained in these experiments suggest that soil resuspension was a significant mechanism for plant contamination and that mulching was effective in reducing that contamination.

Agriculture↗

Solid-phase microextraction to predict bioavailability and accumulation of organic micropollutants in terrestrial organisms after exposure to a field-contaminated soil.

The risk posed by soil contaminants strongly depends on their bioavailability. In this study, a partition-based sampling method was applied as a tool to estimate bioavailability in soil. The accumulation of organic micropollutants was measured in two earthworm species (Eisenia andrei and Aporrectodea caliginosa) and in 30-microm poly(dimethylsiloxane) (PDMS)-coated solid-phase micro extraction (SPME) fibers after exposure to two field-contaminated soils. Within 10 days, steady state in earthworms was reached, and within 20 days in the SPME fibers. Steady-state concentrations in both earthworm species were linearly related to concentrations in fibers over a 10,000-fold range of concentrations. Measured concentrations in earthworms were compared to levels calculated via equilibrium partitioning theory and total concentrations of contaminants in soil. In addition, freely dissolved concentrations of contaminants in pore water, derived from SPME measurements, were used to calculate concentrations in earthworms. Measured concentrations in earthworms were close to estimated concentrations from the SPME fiber measurements. Freely dissolved concentrations of contaminants in pore water, derived from SPME measurements, were used to calculate bioconcentration factors (BCF) in earthworms. A plot of log BCFs against the octanol-water partition coefficient (log Kow) was linear up to a log Kow of 8. These results show that measuring concentrations of hydrophobic chemicals in a PDMS-coated fiber represents a simple tool to estimate internal concentrations of chemicals in biota exposed to soil.

Animals↗

Kinetic desorption and sorption of U(VI) during reactive transport in a contaminated Hanford sediment.

Column experiments were conducted to investigate U(VI) desorption and sorption kinetics in a sand-textured, U(VI)-contaminated (22.7 micromol kg(-1)) capillary fringe sediment from the U.S. Department of Energy (DOE) Hanford site. Saturated column experiments were performed under mildly alkaline conditions representative of the Hanford site where uranyl-carbonate and calcium-uranyl-carbonate complexes dominate aqueous speciation. A U(VI)-free solution was used to study contaminant U(VI) desorption in columns where different flow rates were applied. Sorbed, contaminant U(VI) was partially labile (11.8%), and extended leaching times and water volumes were required for complete desorption of the labile fraction. Uranium-(VI) sorption was studied after the desorption of labile, contaminant U(VI) using different U(VI) concentrations in the leaching solution. Strong kinetic effects were observed for both U(VI) sorption and desorption, with half-life ranging from 8.5 to 48.5 h for sorption and from 39.3 to 150 h for desorption. Although U(VI) is semi-mobile in mildly alkaline, subsurface environments, we observed substantial U(VI) adsorption, significant retardation during transport, and atypical breakthrough curves with extended tailing. A distributed rate model was applied to describe the effluent data and to allow comparisons between the desorption rate of contaminant U(VI) with the rate of shortterm U(VI) sorption. Desorption was the slower process. We speculate that the kinetic behavior results from transport or chemical phenomena within the phyllosilicate-dominated fine fraction present in the sediment. Our results suggest that U(VI) release and transport in the vadose zone and aquifer system from which the sediment was obtained are kinetically controlled.

Carbonates↗

Determination of microbial carbon sources in petroleum contaminated sediments using molecular 14C analysis.

Understanding microbial carbon sources is fundamental to elucidating the role of microbial communities in carbon cycling and in the biodegradation of organic contaminants. Because the majority of anthropogenic contaminants are either directly or indirectly derived from fossil fuels that are devoid of 14C, radiocarbon can be used as a natural inverse tracer of contaminant carbon in the contemporary environment. Here, 14C analysis of individual microbial phospholipid fatty acids (PLFA) was used to characterize the carbon sources utilized bythe active microbial community in salt marsh sediments contaminated by the Florida oil spill of 1969 in Wild Harbor, West Falmouth, MA. A specific goal was to determine whether this community is actively degrading petroleum residues that persist in these sediments. The delta14C values of microbial PLFA in all sediment horizons (contaminated and noncontaminated) matched the delta14C of the total sedimentary organic carbon after petroleum removal, indicating that no measurable metabolism of petroleum residues was occurring. This result agrees with ancillary data such as the delta13C content and distribution of PLFA, and the residual hydrocarbon composition determined by comprehensive two-dimensional gas chromatography (GCxGC) analysis. We hypothesize that microbes have chosen to respire the natural organic matter rather than the residual petroleum hydrocarbons because the former is more labile. Future efforts directed at determining indices of microbial degradation of petroleum hydrocarbons should consider competition with natural organic matter.

Accidents↗