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Cross-contamination of lettuce with Escherichia coli O157:H7.

Contamination of produce by bacterial pathogens is an increasingly recognized problem. In March 1999, 72 patrons of a Nebraska restaurant were infected with enterohemorrhagic Escherichia coli (EHEC) O157:H7, and shredded iceberg lettuce was implicated as the food source. We simulated the restaurant's lettuce preparation procedure to determine the extent of possible EHEC cross-contamination and growth during handling. EHEC inoculation experiments were conducted to simulate the restaurant's cutting procedure and the subsequent storage of shredded lettuce in water in the refrigerator. All lettuce pieces were contaminated after 24 h of storage in inoculated water (2 x 10(9) CFU of EHEC per 3 liters of water) at room temperature or at 4 degrees C; EHEC levels associated with lettuce increased by > 1.5 logs on the second day of storage at 4 degrees C. All lettuce pieces were contaminated after 24 h of storage in water containing one inoculated lettuce piece (approximately 10(5) CFU of EHEC per lettuce piece) at both temperatures. The mixing of one inoculated dry lettuce piece with a large volume of dry lettuce, followed by storage at 4 degrees C or 25 degrees C for 20 h resulted in 100% contamination of the leaves tested. Microcolonies were observed on lettuce stored at 25 degrees C, while only single cells were seen on leaves stored at 4 degrees C, suggesting that bacterial growth had occurred at room temperature. Three water washes did not significantly decrease the number of contaminated leaves. Washing with 2,000 mg of calcium hypochlorite per liter significantly reduced the number of contaminated pieces but did not eliminate contamination on large numbers of leaves. Temperature abuse during storage at 25 degrees C for 20 h decreased the effectiveness of the calcium hypochlorite treatment, most likely because of bacterial growth during the storage period. These data indicate that storage of cut lettuce in water is not advisable and that strict attention must be paid to temperature control during the storage of cut lettuce.

Calcium Compounds↗

Visible contamination on animals and carcasses and the microbiological condition of meat.

It is generally assumed that preventing visible contamination of or removing visible contamination from carcasses will enhance the microbiological safety of meat. Visible contamination of carcasses can be reduced by washing or otherwise cleaning animals before slaughter, by dehairing hides before carcasses are skinned or dressed with the skin on, or by performing skinning and eviscerating operations in manners that avoid the transfer of filth from the hide to the meat or the spillage of gut contents. Visible contamination can be removed by washing, trimming, or vacuuming carcasses. The available data appear to indicate that, of the various actions that can be taken to obtain carcasses that are free of visible contamination, only minimizing the visible contamination of meat during skinning and eviscerating operations may also ensure a degree of control over the microbiological contamination of meat. It might be preferable for visible contamination to be controlled largely by superior skinning and eviscerating practices rather than by animal or carcass cleaning treatments, which may not prevent the depositing of bacteria on or the removal of substantial numbers of bacteria from carcasses.

Abattoirs↗

Persistence of enterohemorrhagic Escherichia coli O157:H7 in soil and on leaf lettuce and parsley grown in fields treated with contaminated manure composts or irrigation water.

Outbreaks of enterohemorrhagic Escherichia coli O157:H7 infections associated with lettuce and other leaf crops have occurred with increasing frequency in recent years. Contaminated manure and polluted irrigation water are probable vehicles for the pathogen in many outbreaks. In this study, the occurrence and persistence of E. coli O157:H7 in soil fertilized with contaminated poultry or bovine manure composts or treated with contaminated irrigation water and on lettuce and parsley grown on these soils under natural environmental conditions was determined. Twenty-five plots, each 1.8 by 4.6 m, were used for each crop, with five treatments (one without compost, three with each of the three composts, and one without compost but treated with contaminated water) and five replication plots for each treatment. Three different types of compost, PM-5 (poultry manure compost), 338 (dairy manure compost), and NVIRO-4 (alkaline-stabilized dairy manure compost), and irrigation water were inoculated with an avirulent strain of E. coli O157:H7. Pathogen concentrations were 10(7) CFU/g of compost and 10(5) CFU/ml of water. Contaminated compost was applied to soil in the field as a strip at 4.5 metric tons per hectare on the day before lettuce and parsley seedlings were transplanted in late October 2002. Contaminated irrigation water was applied only once on the plants as a treatment in five plots for each crop at the rate of 2 liters per plot 3 weeks after the seedlings were transplanted. E. coli O157:H7 persisted for 154 to 217 days in soils amended with contaminated composts and was detected on lettuce and parsley for up to 77 and 177 days, respectively, after seedlings were planted. Very little difference was observed in E. coli O157:H7 persistence based on compost type alone. E. coli O157:H7 persisted longer (by > 60 days) in soil covered with parsley plants than in soil from lettuce plots, which were bare after lettuce was harvested. In all cases, E. coli O157:H7 in soil, regardless of source or crop type, persisted for > 5 months after application of contaminated compost or irrigation water.

Disease Outbreaks↗

[Bacterial contamination of multi-dose ocular solutions. A prospective study at the Grenoble Teaching Hospital].

The bacterial contamination rate of multidose ocular solutions used by hospitalized patients was evaluated by culturing vial dropper tips and residual solution in vials. Bacterial colonies were counted and identified. Overall 39 (23.5%) selected vials were contaminated. Contamination rates were 17.7% (20/113) for vials used by ophthalmology ward patients and 35.8% (19/53) for vials used by internal medicine and gerontology patients (P < 0.02). The most commonly identified organisms were part of the normal commensal flora. Three ophthalmology patients were using vials contaminated with Pseudomonas aeruginosa. A significant (P < 0.01) positive correlation was found between vial contamination rate and duration of vial use. Vials containing an antimicrobial agent were less likely to be contaminated than vials without antimicrobials (P < 0.01). No clinical consequences of vial contamination were identified. However, ocular solution vial contamination carries a risk of infection. Our data are evidence of inadequate efficacy of preservatives present in ocular solutions. The standard practice of using ocular solution vials for seven days in health care facilities may need to be reappraised. Care should be taken to ensure that health care providers and patients understand the rules for ocular solution use. Unit-dose presentations may be preferable over multi-dose presentations for in hospital treatment.

Anti-Bacterial Agents↗

Expansion of contaminated amalgams assessed by photoelastic resin.

OBJECTIVE: Eight amalgam alloys, 6 high-copper and 2 low-copper, were assessed for expansion via photoelastic resin. METHOD AND MATERIALS: The photoelastic resin sheet was sectioned into 20 x 25-mm blocks, and two 4 x 8-mm holes were drilled into the resin. The amalgam alloys were hand condensed into the prepared holes. Test conditions for each alloy were (1) uncontaminated, (2) contaminated with 5 microL of Ringer's solution, and (3) contaminated with 5 microL of cell culture medium. RESULTS: Contaminated high-copper amalgam alloys may exhibit expansion to varying degrees but do not show the classic delayed expansion. One spherical, nonzinc high-copper alloy showed no expansion, whether it was uncontaminated or contaminated, and 3 high-copper alloys with different amounts of zinc (0% to 1%) showed slight expansion when contaminated. One low-copper alloy displayed delayed expansion; within 17 days after initial expansion in the photoelastic resin, it had exceeded the greatest stress observed in any contaminated, high-copper zinc-containing alloy at 3 months. CONCLUSION: Contamination of dental amalgam is to be avoided, but a zinc-containing high-copper amalgam will not exhibit classic delayed expansion even if contaminated.

Birefringence↗

[The existence of contamination in enzymes used for the isolation of Aspergillus and Candida species DNA].

Molecular biological methods which are widely used in different fields, have been replaced with conventional diagnostic tests for the early diagnosis of invasive fungal infections, recently. Polymerase chain reaction (PCR) is one of these methods, with high specificity and sensitivity, which is accepted throughout the world. However, the enzymes that are used for the isolation of target DNA, may be contaminated with the gene sequences of some other fungal species in the preparation steps and may affect the PCR results. The studies showed that, the contamination of these enzymes with any type of fungi during their production steps, leads false positive results in PCR tests. So, the additional studies are recommended for minimizing the contamination. The aim of this study was to search whether the enzymes necessary for the isolation of fungal target DNA, used in PCR, are contaminated or not. For this purpose, five different enzymes namely, Zymolase 20T (from Arthrobacter luteus, two examples from different companies), lyticase (from Arthrobacter luteus), lysing enzyme (from Trichoderma harzianum) and proteinase K (from Tritrachium album) have been investigated for the presence of contamination. As a result, both of zymolase 20T enzymes were found to be contaminated from some fungal species with the demonstration of 18S rRNA gene sequences, while the other enzymes were found non-contaminated. By the help of this method, the most suitable enzyme for PCR was chosen and fungal contamination was prevented.

Aspergillus fumigatus↗

[Influence of milking technique, milking hygiene and environmental hygiene parameters on the microbial contamination of milking machines].

It was the aim of this study to investigate the effect of various factors of the milking technique, milking hygiene and environment on microbial contamination of the milking machine. In 31 dairy herds, the degree of bacterial contamination was examined by taking swabs at four locations (teat cup liner, claw, short and long milk tube) before the milking procedure was started using a standardized protocol (DIN ISO 6887-1:1999). Furthermore, the total germ count was determined in the first milk entering the bulk tank as well as in the bulk tank milk following milking. For each farm, the quality of the milking process and the condition of the milking machine as well as of various environmental factors were recorded. A subjective evaluation of the status of the milking cluster or other parts of the milking machine ("good" or "moderate-poor") gave more information about bacterial contamination than the determination of age and type of material used. A temperature of the rinsing water of < 42 degrees C increased the contamination with Pseudomonas spp. and coliform bacteria. Milking clusters kept out of the cluster pick-up between milking had a higher risk of microbial contamination. Various methods of teat cleaning before milking or of postmilking teat disinfection did not affect the contamination of the milking machine and the bulk tank milk with environmental bacteria. Furthermore, type of bedding material affected bacterial contamination of milking clusters and bulk tank milk. In conclusion, our results suggest that the microbial contamination of the milking machine is not only influenced by the sanitation pro-

Animals↗

Relationship between length of time and contamination in open intravenous solutions.

This study investigated rate of contamination in simulated in-use intravenous solutions to obtain data from which to recommend a standard time period for the administration of intravenous solutions. Although the Center for Disease Control and many medical institutions have recommended that all bottles of in-use intravenous solutions be changed every 24 hours, there are no published data to support the 24-hour standard. In this study, samples were drawn from 80 bottles of Ringer's Lactate solution at predetermined time intervals over 48 hours and cultured on blood agar plates to determine rate of contamination. A low incidence of intravenous solution contamination was found at all time periods. Of 430 total samples, only 12 (2.8%) were contaminated. Results of this study did not indicate a significant relationship between time and contamination of simulated in-use intravenous solutions. Therefore, contamination that does occur within 48 hours in intravenous solutions must be introduced from some exogenous source. Further research is required to elucidate exogenous sources of contamination. In the meantime, it can be concluded, the 24-hour standard is adequate to protect against contamination that would occur solely from preparation of the intravenous solution for administration.

Drug Contamination↗

[Contamination and secondary reactions in the processing of dietary fats].

The contamination of dietary fats and oils can have various origins: contamination of the raw materials that could not be eliminated by the industrial process; refining completely eliminates this type of contamination; contamination with one of the processing aids can occur if the process is not properly conducted; contamination with packaging materials will increase with the storage duration of the finished product (in the case of vinyl chloride monomer and styrene contamination); only the strict control and adequate choice of the materials and packaging conditions will allow complete elimination of this contamination type. The presence of small amounts of another fat (in vegetable margarines for instance) should also be mentioned although it is not a real contamination. The secondary reactions that may occur during the processing of dietary fats are primarily due to unwanted chemical reactions leading to the formation of fatty acid or native glyceride isomers (e.g. formation of geometric isomers during deodorization, formation of position isomers during hydrogenation). In the present stage of knowledge, it is fairly easy to avoid secondary reactions during deodorization, those that take place during hydrogenation are not yet under control, however.

Chemical Phenomena↗

Contaminated stethoscopes revisited.

BACKGROUND: Because of their universal use by medical professionals, stethoscopes can be a source of nosocomial infections. OBJECTIVE: To determine the frequency of contamination of stethoscopes with bacteria and fungi. METHODS: Cultures were obtained from 200 stethoscopes from four area hospitals and outpatient clinics in Houston, Tex. The frequency of stethoscope contamination in different groups of hospital personnel and medical settings was determined. We also measured the frequency of antimicrobial resistance of the staphylococcal strains that were isolated. RESULTS: One hundred fifty-nine (80%) of the 200 stethoscopes surveyed were contaminated with microorganisms. The majority of organisms that were isolated were gram-positive bacteria, primarily Staphylococcus species. Fifty-eight percent of the Staphylococcus species that were isolated, including four (17%) of 24 Staphylococcus aureus isolates, were resistant to methicillin. Physicians' stethoscopes were contaminated more often than those of other medical personnel groups (P = .02). Stethoscopes used only in designated areas were contaminated less frequently than stethoscopes belonging to individual medical personnel (P = .01). Although stethoscopes were contaminated in all areas, stethoscopes from the pediatric medical setting were contaminated less frequently than those from other hospital areas (P = .009). CONCLUSIONS: Stethoscope use may be important in the spread of infectious agents, including antimicrobial-resistant strains, and strategies to reduce the contamination of stethoscopes should be developed. We recommend disinfection of stethoscopes or regular use of disposable stethoscope covers.

Auscultation↗

Survival of contaminating bacteria in over-the-counter artificial tears.

BACKGROUND: The use of OTC artificial tears is becoming increasingly widespread. Unintentional contamination of artificial tear products potentiates a risk of developing bacterial keratitis or exacerbating an existing eye infection. Presently, no published study has investigated recovery from bacterial contamination of artificial tear products. METHODS: Four different over-the-counter (OTC) artificial tear products were contaminated with two types of bacteria. Staphylococcus aureus and pseudomonas aeruginosa were analyzed for the survivability of the microorganisms at various times following contamination. RESULTS: In this cross-sectional study, it was found that all the preserved brands showed a significant recovery rate from contamination compared to the preservative-free brand. After 9 hours, the preservative-free brand did not completely regain sterility after contamination with either bacteria. CONCLUSIONS: This study indicates that preserved artificial tears recover from bacterial contamination at a significantly faster rate than preservative-free products. It is likely that the recovery from bacterial contamination is not solely attributed to the preservative, but that other variables may contribute.

Colony Count, Microbial↗

Airborne contamination during blow-fill-seal pharmaceutical production.

The routes of airborne contamination, during Blow-Fill-Seal (BFS) production, were studied using tracer gas, particles and bacteria. The prevention of airborne contamination, by the air shower at the point of fill, was effective (> 99.2% efficient). However, microbe-carrying particles could gain access, by deposition or air exchange, when the containers were cut open and before they shuttled under the protection of the air shower. The use of SF6 tracer gas demonstrated that when the air shower was not on, 50% of the air within the containers came from the area round the machine. When the air shower was switched on, only about 5% of the air came from the surroundings. Airborne microbial contamination of containers is in proportion to: the number of airborne microbes around the machine, the time the container is open, the neck area and the amount of air left within the container. The likely microbial contamination rate can be calculated from a model incorporating these variables. Microbial contamination of containers during BFS manufacturing is normally very low, but by increasing the naturally occurring bacteria in the air of the production rooms by about 100-fold, it was possible to verify the accuracy of this model. The contamination model agrees well with the observation that microbial contamination levels of between 1 in 10(5) and in 10(7) will be found when small containers (< 10 ml) are filled in conventionally ventilated rooms. To achieve similar contamination rates when filling of larger bottles, it is likely that unidirectional flow, or barrier technology will be required.

Air Microbiology↗

Transplantation of microbially contaminated cadaver kidneys.

Bacterial cultures of graft perfusion fluid were taken on 514 renal perfusions during a ten-year period. A total of 22 positive cultures were found. Two sets of kidneys were discarded because of the bacteriology report. Only two possible wound infections were found that could be attributable to the preoperative contamination even though 19 of 24 patients with "contaminated" kidneys received no antibiotics. No loss of kidneys or life was found. Of the 22 contaminated perfusates, 18 (80%) originated from other institutions although only 144 (28%) of the total kidneys that were perfused came from other institutions. The majority of cultures indicated that the organisms were skin contaminants or organisms of low virulence. Microbial contamination of perfused cadaver kidneys is uncommon (1.2% in local kidneys and 9.3% in shared kidneys). Contaminated kidneys are not a major source of infection in renal transplant recipients. Kidneys that are contaminated with frequently virulent Gram-negative bacteria, such as Escherichia coli, should be discarded. Contamination of kidneys with Gram-positive organisms should not be a contraindication for transplantation, but the patients should probably be given prophylactic antibiotics.

Cadaver↗

NO(x) contamination in laboratory ware and effect of countermeasures.

Contamination of various types of laboratory wares with NO(x) (NO(-)(2) and NO(-)(3)) was assessed systematically and the effect of extensive washing as a countermeasure was evaluated. Mean NO(x) contamination arising from a model procedure for NO(x) determination in plasma was 0.93 microM (range, 0.35-1.49 microM). The major source of contamination included conical tubes (54.8%) and pipette tips used for transfer of solution (12.3-16.3%). Except for soft glassware, most NO(x) contamination could be washed out by pure water. Although NO(x) contamination in respective laboratory wares could be reduced below detection levels by extensive washing, summation of the contamination through the model procedure could not be completely abolished (but the effect of washing persisted at least 10 days). Heavy contamination was noted in glassware (especially soft glass) and ultrafiltration units, which was difficult to remove. Several types of vacuum blood sampling tubes contained various levels of NO(x). Our results indicated that a small but significant amount of contamination remained in laboratory ware even after extensive washing, and that it is advisable to avoid the use of glassware (soft glass), ultrafiltration units, and vacuum blood sampling tubes during the processing of clinical sampling for the measurement of NO(x).

Blood Specimen Collection↗

Effect of saliva contamination on shear bond strength of orthodontic brackets when using a self-etch primer.

The effect of saliva contamination on the shear bond strength of orthodontic brackets, at various stages of the bonding procedure using a new self-etch primer was studied. The samples were divided into 4 groups according to 4 different enamel surface conditions: Group A: dry; Group B: saliva contamination before priming; Group C: saliva contamination after priming, and Group D: saliva contamination before and after priming. Stainless steel brackets were bonded in each test group with a light-cured composite resin (TransbondXT 3M). The shear bond strength was determined in the first 30 min after bonding. The analysis of variance indicated that the shear bond strengths of the 4 groups were significantly different (F = 11.89, P < 0.05). Tukey HSD tests indicated that contamination both before and after the application of the acid-etch primer resulted in a significantly lower (=4.6 +/- 1.7 MPa) shear bond strength than either the control group (=8. 8 +/- 1.9 MPa) or the groups where contamination occurred either before ( = 7.9 +/- 2.0 MPa) or after (= 6. 9 +/- 1.5 MPa) the application of the primer. It was concluded that the new acid-etch primer could maintain adequate shear bond strength if contamination occurred either before or after the application of the primer. On the other hand, contamination both before and after the application of the primer could significantly reduce the shear bond strength of orthodontic brackets.

Acid Etching, Dental↗

Mineralization of clapper rail eggshell from a contaminated salt marsh system.

The effect of contamination on eggshell mineralization has been studied for clapper rails (Rallus longirostris) inhabiting a contaminated salt marsh in coastal Georgia. To assess the impact of contaminants, the thickness, microstructure (crystal orientation), mineral composition, and chemistry of shell material were analyzed from a contaminated site and a nearby reference site using optical microscopy, X-ray diffraction, inductively coupled plasma mass spectrometry, and gas chromatography with electron capture detector. Eggshells from the contaminated site were generally thinner than those from the reference site. Also, eggshells from the contaminated site were abnormally brittle and contained anomalous microstructural attributes. The combination of reduced shell thickness and anomalous microstructure resulted in weaker eggshells, which in turn could pose a significant threat to the reproductive success of the affected population.PCB concentrations in eggshells were at background levels in both sites. Eggshells from the contaminated site had higher concentrations of heavy metals, specifically mercury, than the reference site. The structural changes observed in eggshells may be related to the concentration of specific metals ( e.g., Mg, Cu, Zn, Pb, and Hg) in shell, however, statistical analyses indicated that metals only explained a small portion of the observed variation in properties ( i.e., thickness, crystal orientation). Further analysis is required to better constrain the factors leading to unusually weak eggshells in the contaminated site.

Animals↗

Contaminant residues in snapping turtle (Chelydra s. serpentina) eggs from the Great Lakes-St. Lawrence River basin (1999 to 2000).

To determine temporal and geographic variations of environmentally persistent pollutants, the concentrations of several contaminants were measured in snapping turtle eggs in 1999 and 2000. Contaminants included polychlorinated biphenyls (PCBs), non- ortho PCBs, polychlorinated dibenzodioxins and furans, organochlorine pesticides, and total mercury. Egg collection sites within the St. Lawrence Area of Concern (AOC) and the Territory of Akwesasne included Turtle Creek, Snye Marsh, and Raquette River. Additional contaminated sites were Gray's Creek (within the St. Lawrence AOC boundary), Coote's Paradise (Hamilton Harbour AOC), and the Territory of Walpole Island (St. Clair AOC). Reference sites were Lake Sasajewun, Algonquin Provincial Park in central Ontario, and in close proximity to the St. Lawrence Area AOC, Hoasic Creek and Cooper Marsh. Eggs from clutches collected from Akwesasne contained the highest concentrations of contaminants. Mean total PCB concentrations for Akwesasne clutches ranged from 1,900 to 61,000 ng/g wet weight (w.w.), which was 360 to 3,100 times higher than the mean concentrations at the reference sites. The high levels of contaminants in eggs, from sites within Akwesasne, were consistent with trends reported in the past and reflect the influence of the proximity of heavy industry. Current contaminant concentrations in clutches from Coote's Paradise are comparable with those recorded in 1989 and 1990 but are 200 to 800 times lower than concentrations reported in 1984 and 1988. Eggs from Lake Sasajewun contained relatively low contaminant levels, which is consistent with atmospheric deposition. Our results support that snapping turtles are good indicators of temporal and geographic contaminant trends.

Animals↗

Increased abundance of terrestrial isopod populations in terrestrial ecosystems contaminated with petrochemical wastes.

Arthropods are integral components of the food chain, and because many reside in close association to the soil and its contaminants, their communities may be sensitive indicators of ecotoxicity. We examined the influence of petrochemical contaminants on the abundance of several taxonomic groups of macroarthropods, with an emphasis on isopods, residing on two reference sites and three petrochemical-contaminated sites at an abandoned oil refinery site in southwestern Oklahoma. Relative densities of surface-dwelling macroarthropod assemblages were significantly greater on contaminated sites than reference sites. Differences in terrestrial isopod populations were particularly remarkable and consistent across study sites, with densities approaching 180-fold greater on contaminated than reference sites. Similarity comparisons indicated that abundances and composition of macroarthropod assemblages sampled on the contaminated sites were more similar to one another than to the communities residing on reference sites. Although preliminary, these initial results support the hypothesis that macroarthropod assemblages, in particular isopods, are sensitive to soil contaminants and could be a valuable approach to evaluating the effects of petrochemical contamination of soils on terrestrial ecosystems.

Animals↗