Fecal Coliform Methods for Examination of Sea Water: Interlaboratory Evaluation of Split Sample Analysis

1978 ◽  
Vol 61 (4) ◽  
pp. 772-778
Author(s):  
John J Miescier ◽  
Virgil E Carr ◽  
John F Musselman ◽  
Santo A Furfari

Abstract An interlaboratory study was conducted to compare the effectiveness of the following 3 multiple-tube fermentation methods for determining the most probable number (MPN) of Escherichia coli in a split artificial sea water sample: ( 1 ) the 72-hr standard methods procedure of the American Public Health Association, (2) a 24-hr elevated-temperature test using A-l medium, and (3) a 24-hr elevated temperature test modified to include an initial 3-hr resuscitation period using A-l medium. The capability of the laboratories to perform the 3 test procedures was also compared. Split sample replicates with low, medium, and high levels of E. coli were examined in 18 laboratories in the United States and Canada. Data indicate that the laboratories performed each test with equal capability, and all 3 procedures were equally effective in enumerating the strain of E. coli used in this investigation. By virtue of its homogeneity and stability, the split sample served as an appropriate specimen for this study and could probably be used as a proficiency test specimen for evaluating laboratory analyst performance in the bacteriological examination of sea water.

2010 ◽  
Vol 93 (2) ◽  
pp. 576-586 ◽  
Author(s):  
Erin Crowley ◽  
Patrick Bird ◽  
Marianne Torontali ◽  
Katherine Goetz ◽  
James Agin ◽  
...  

Abstract The automated method for enumeration of Escherichia coli, TEMPO<sup/> EC, in foods uses a dehydrated culture medium and enumeration card containing 48 wells across three different dilutions for the automatic determination of the most probable number (MPN). The alternative method was compared in a multilaboratory collaborative study to AOAC Official MethodSM 966.24. Six food types were artificially contaminated with E. coli: raw ground beef, bagged lettuce, cooked chicken, pasteurized crabmeat, frozen green beans, and pasteurized whole milk. All foods were analyzed for E. coli counts by 11 collaborating laboratories throughout the United States. Test portions from the six food types each contaminated at four different contamination levels were evaluated. The study demonstrated that the TEMPO EC method is a reliable, automated assay for the enumeration of E. coli in foods.


2018 ◽  
Vol 81 (3) ◽  
pp. 400-411 ◽  
Author(s):  
Guodong Zhang ◽  
Yi Chen ◽  
Lijun Hu ◽  
David Melka ◽  
Hua Wang ◽  
...  

ABSTRACT The objective of this research was to assess the microbiological status of leafy greens, sprouts, and melons from U.S. markets. A total of 14,183 samples of leafy greens, 2,652 samples of sprouts, and 3,411 samples of melons were collected throughout the United States from 2009 to 2014. The samples were analyzed for aerobic plate counts, total coliform counts, Escherichia coli counts, and the presence and levels of Salmonella, Shigella, Listeria monocytogenes, and Shiga toxin–producing E. coli (STEC), depending on the year and type of produce. Among the leafy greens, no E. coli O157:H7 or non-O157 STEC were detected from iceberg lettuce samples. The overall prevalences of Salmonella, E. coli O157:H7, non-O157 STEC, and L. monocytogenes in the 14,183 samples of leafy greens were 0.05, 0.01, 0.07, and 0.11%, respectively. Among sprout samples, no Salmonella or E. coli O157:H7 was detected, and the overall prevalences of non-O157 STEC and L. monocytogenes were 0.04 and 0.11%, respectively. Among melon samples, no Salmonella was detected from cucumbers, no L. monocytogenes was detected from cantaloupes, and the overall prevalences of Salmonella and L. monocytogenes were 0.12 and 0.23%, respectively. L. monocytogenes levels were 0.4 to 1,470 most probable number (MPN)/g in leafy greens, 0.36 to 1,100 MPN/g in sprouts, and <0.03 to 150 MPN/g in melons, and most positive samples had low levels of these pathogens. The isolates from these foods were very diverse genetically. Foodborne pathogens, including Salmonella, STEC, and L. monocytogenes, had relatively low prevalences in the produce surveyed. Because these foods are usually consumed raw, measures should be taken to significantly minimize the presence and levels of human pathogens.


1999 ◽  
Vol 122 (2) ◽  
pp. 185-192 ◽  
Author(s):  
J. TUTTLE ◽  
T. GOMEZ ◽  
M. P. DOYLE ◽  
J. G. WELLS ◽  
T. ZHAO ◽  
...  

Between November 1992 and February 1993, a large outbreak of Escherichia coli O157[ratio ]H7 infections occurred in the western USA and was associated with eating ground beef patties at restaurants of one fast-food chain. Restaurants that were epidemiologically linked with cases served patties produced on two consecutive dates; cultures of recalled ground beef patties produced on those dates yielded E. coli O157[ratio ]H7 strains indistinguishable from those isolated from patients, confirming the vehicle of illness. Seventy-six ground beef patty samples were cultured quantitatively for E. coli O157[ratio ]H7. The median most probable number of organisms was 1·5 per gram (range, <0·3–15) or 67·5 organisms per patty (range, <13·5–675). Correlation of the presence of E. coli O157[ratio ]H7 with other bacterial indicators yielded a significant association between coliform count and the presence of E. coli O157[ratio ]H7 (P=0·04). A meat traceback to investigate possible sources of contamination revealed cattle were probably initially colonized with E. coli O157[ratio ]H7, and that their slaughter caused surface contamination of meat, which once combined with meat from other sources, resulted in a large number of contaminated ground beef patties. Microbiological testing of meat from lots consumed by persons who became ill was suggestive of an infectious dose for E. coli O157[ratio ]H7 of fewer than 700 organisms. These findings present a strong argument for enforcing zero tolerance for this organism in processed food and for markedly decreasing contamination of raw ground beef. Process controls that incorporate microbiological testing of meat may assist these efforts.


2009 ◽  
Vol 75 (23) ◽  
pp. 7417-7425 ◽  
Author(s):  
H. N. Chinivasagam ◽  
T. Tran ◽  
L. Maddock ◽  
A. Gale ◽  
P. J. Blackall

ABSTRACT This study assessed the levels of two key pathogens, Salmonella and Campylobacter, along with the indicator organism Escherichia coli in aerosols within and outside poultry sheds. The study ranged over a 3-year period on four poultry farms and consisted of six trials across the boiler production cycle of around 55 days. Weekly testing of litter and aerosols was carried out through the cycle. A key point that emerged is that the levels of airborne bacteria are linked to the levels of these bacteria in litter. This hypothesis was demonstrated by E. coli. The typical levels of E. coli in litter were ∼108 CFU g−1 and, as a consequence, were in the range of 102 to 104 CFU m−3 in aerosols, both inside and outside the shed. The external levels were always lower than the internal levels. Salmonella was only present intermittently in litter and at lower levels (103 to 105 most probable number [MPN] g−1) and consequently present only intermittently and at low levels in air inside (range of 0.65 to 4.4 MPN m−3) and once outside (2.3 MPN m−3). The Salmonella serovars isolated in litter were generally also isolated from aerosols and dust, with the Salmonella serovars Chester and Sofia being the dominant serovars across these interfaces. Campylobacter was detected late in the production cycle, in litter at levels of around 107 MPN g−1. Campylobacter was detected only once inside the shed and then at low levels of 2.2 MPN m−3. Thus, the public health risk from these organisms in poultry environments via the aerosol pathway is minimal.


1998 ◽  
Vol 61 (7) ◽  
pp. 903-906 ◽  
Author(s):  
STEVEN PAO ◽  
G. ELDON BROWN

Citrus fruit surface microbial populations were evaluated following various packingline processes of seven Florida commercial packinghouses. At each packinghouse, six fruits (oranges or tangerines) were collected at each of four sampling points. The sampling was conducted in duplicate; thus, 336 fruit were evaluated during this survey. Average aerobic plate counts and yeast and mold counts on fruit surfaces before washing were about 4.0 log CFU/cm2 and 3.3 log CFU/cm2, respectively, and were reduced to 2.1 log CFU/cm2 and 1.3 log CFU/cm2, respectively, by packinghouse processing. Waxing alone reduced the average fruit surface aerobic plate counts and coliform counts from 3.7 log CFU/cm2 and 35.2 most probable number (MPN)/cm2, respectively, to 2.6 log CFU/cm2 and 1.4 MPN/cm2. No Escherichia coli was recovered from fruit at the end of packinghouse processing, and no salmonellae were found on fruit during the entire processing. In an inoculation study to test the effect of packinghouse processes, test organism E. coli was applied to fruit to achieve a high level (4.8 log CFU/cm2) of contamination. The average E. coli count was reduced about 2.4 log cycles by washing and rinsing with potable water (40 psi, 25 °C) for about 30 s. The combination of washing and waxing significantly reduced the inoculated level of E. coli from 4.8 to 1.4 log CFU/cm2.


Author(s):  
YOJANA Y. PATIL ◽  
VAISHNVI B. SUTAR ◽  
ARPITA P. TIWARI

Objective: The present study was aimed at the biological synthesis of magnetic iron nanoparticles by using the plant extract of Tridax procumbens and also to study their antimicrobial property against gram-negative bacteria (Escherichia coli). Methods: The synthesis of magnetic iron nanoparticles was carried out by the co-precipitation method using biological methods like plant extract as reducing agent and capping agents are biocompatible and non-hazardous. These nanoparticles were characterized by UV-Visible spectroscopy, XRD (X-Ray Diffraction), and SEM (Scanning Electron Microscope). As well as antibacterial activity of the nanoparticles was carried out by agar well diffusion method and Most Probable Number (MPN) method against gram-negative E. coli (Escherichia coli) bacteria. Results: The average crystallite size of Magnetic Nanoparticles (MNPs) was found to be 72 nm by X-ray diffraction. The optical absorption band at wavelengths of 240 nm and 402 nm was obtained from the UV Visible spectrum. Spherical shape morphology was observed in SEM studies. The antibacterial assay clearly expressed that E. coli showed a maximum zone of inhibition (15±0.15 mm) at 2 mg/ml and 1 mg/ml concentration was found for Magnetic Nanoparticles. In the Most Probable Number (MPN) test it is seen that the bacterial count is reduced after adding synthesized NPs into the water sample. Conclusion: The results of the present study conclude that the Magnetic Nanoparticles synthesized using Tridax procumbens leaf extracts is found to be stable and show good antibacterial activity against gram-negative (Escherichia coli) bacteria.


2019 ◽  
Vol 2 (2) ◽  
pp. a13-19
Author(s):  
ELEXSON NILLIAN ◽  
AMIZA NUR ◽  
DIYANA NUR ◽  
AMIRAH ZAKIRAH ◽  
GRACE BEBEY

Contamination of drinks with E. coli O157:H7 served in food premises such as restaurants can cause haemorrhagic colitis and haemolytic uremic syndrome to humans. The presence or absence of faecal pathogen was demonstrated using coliform group as indicator microorganisms. Therefore, this study was conducted to detect the presence of E. coli O157:H7 in drinking water from food restaurant premise in Kota Samarahan and Kuching to ensure safe and potable drinking water is served to the consumer. A total of thirty (n=30) drink samples including six types of each of the samples are cold plain water, iced tea, iced milo, syrup and iced milk tea. Most Probable Number (MPN) procedure was used in this study to enumerate the MPN values of coliform bacteria in each drink collected. A total of 53.33% (16/30) of the drink samples showed positive E. coli detection. Then, the PCR assay showed 6.25% (one out of 16 isolates) samples were positive and carried stx1 gene produced by E. coli O157:H7 in iced milo sample types. This study showed the drinks collected from food premises was contaminated with faecal contamination, which was not safe to drink by the consumer. Therefore, preventive actions should be taken to prevent foodborne illness outbreak in future


2013 ◽  
Vol 64 (5) ◽  
pp. 447 ◽  
Author(s):  
Robert J. Wilcock ◽  
Ross M. Monaghan ◽  
Richard W. McDowell ◽  
Piet Verburg ◽  
Jonny Horrox ◽  
...  

A study (2004–11) of a dairy catchment stream entering an oligotrophic lake in an area of very high rainfall (~5 m year–1) yielded median concentrations of total nitrogen (TN), total phosphorus (TP), suspended sediment (SS) and Escherichia coli (E. coli) of 0.584, 0.074 and 3.7 g m–3, and 405/100 mL (most probable number method), respectively. Trend analysis indicated significant (P < 0.01) decreases for TN (–0.08 ± 0.02 g m–3 year–1), TP (–0.01 ± 0.005 g m–3 year–1) and SS (–0.45 ± 0.14 g m–3 year–1) and were partly attributable to improved exclusion of cattle from the stream. Water balance calculations indicated that approximately one-half the rainfall left as deep drainage that by-passed catchment outlet flow recorders. Estimates of catchment yields for TN were improved by taking into account groundwater hydrology and concentrations from well samples. Storm-flow monitoring inflows exceeding the 97.5th percentile contributed ~40% of total loads leaving the catchment so that specific yields for SS, TN and TP augmented by groundwater inputs and storm flows were ~960, 45 and 7 kg ha–1 year–1, respectively. These compared well with modelled results for losses from dairy farms in the catchment of 40–60 kg N ha–1 year–1 and 5–6 kg P ha–1 year–1 and indicated that attenuation losses were relatively small.


2006 ◽  
Vol 4 (3) ◽  
pp. 389-393 ◽  
Author(s):  
Reyneé W. Sampson ◽  
Sarah A. Swiatnicki ◽  
Vicki L. Osinga ◽  
Jamie L. Supita ◽  
Colleen M. McDermott ◽  
...  

A concern for public health officials is the presence of Escherichia coli (E. coli), an indicator of fecal contamination, in monitoring recreational waters. While E. coli is unlikely to cause disease in humans, its presence may indicate other more pathogenic microorganisms. Many factors can lead to changes in the survival of E. coli outside of the animal intestine and may affect the probability of colonizing a new host. Survival of bacteria in recreational water has been linked to water temperature, and most recently to the presence of sand on the beach. This project looked at the survival of an environmental E. coli isolate in lake water. Lake water microcosms were placed at 4, 10, 14, or 25°C for up to 36 d and an enzyme-substrate test (Colisure®, IDEXX Corp.) was used to determine the most probable number (MPN) of E. coli/100 ml water. E. coli numbers at all temperatures declined over the duration of the experiment. The decline was most pronounced at 14°C and was slowest at 4°C. The presence of sand in the microcosm increased the time that E. coli survived, regardless of temperature. From a beach management standpoint, these findings indicate that E. coli may persist in the environment in cooler water longer than in the warmer water encountered in late summer.


Author(s):  
Marie Limoges ◽  
Deborah A. Neher ◽  
Thomas R. Weicht ◽  
Patricia D. Millner ◽  
Manan Sharma ◽  
...  

Composted or heat-treated Biological Soil Amendments of Animal Origin (BSAAO) can be added to soils to provide nutrients for fresh produce. These products lower the risk of pathogen contamination of fresh produce when compared with use of untreated BSAAO; however, meteorological conditions, geographic location, and soil properties can influence the presence of pathogenic bacteria, or their indicators (e.g., generic E. coli) and allow potential for produce contamination. Replicated field plots of loamy or sandy soils were tilled and amended with dairy manure compost (DMC), poultry litter compost (PLC), or no compost (NoC) over two different field seasons, and non-composted heat-treated poultry pellets (HTPP) during the second field season. Plots were inoculated with a three-strain cocktail of rifampicin-resistant E. coli (rE.coli) at levels of 8.7 log CFU/m2. Direct plating and most probable number (MPN) methods measured the persistence of rE.coli and Listeria spp. in plots through 104 days post-inoculation. Greater survival of rE. coli was observed in PLC plots in comparison to DMC plots and NoC plots during year 1 (P &lt; 0.05). Similar trends were observed for year 2, where rE. coli survival was also greater in HTPP amended plots (P &lt; 0.05). Survival of rE. coli was dependent on soil type, where water potential and temperature were significant covariables. Listeria spp. were found in NoC plots, but not in plots amended with HTPP, PLC or DMC. Radish data demonstrate that PLC treatment promoted the greatest level of rE.coli translocation when compared to DMC and NoC treatments (P  &lt; 0.05). These results are consistent with findings from studies conducted in other regions of the US and informs Northeast produce growers that composted and non-composted poultry-based BSAAO supports greater survival of rE. coli in field soils. This result has the potential to impact the food safety risk of edible produce grown in BSAAO amended soils as a result of pathogen contamination.


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