Evaluation of Coli-ID and MUG Plus media for recovering Escherichia coli and other coliform bacteria from groundwater samples

2001 ◽  
Vol 43 (12) ◽  
pp. 213-216 ◽  
Author(s):  
R. A. Sueiro ◽  
M. Araujo ◽  
C. J. Santos ◽  
M. J. Gómes ◽  
M. J. Garrido

Several chromogenic media for detecting coliform bacteria in water are commercially available including Coli ID medium (ID) (bioMérieux) and MUG Plus cefsulodin agar (MP) (Laboratorios Microkit, S.L.). Since little is known about the performance of these media, we have evaluated their usefulness for recovering Escherichia coli and other coliform organisms in groundwaters used for direct human consumption. Variance analysis of obtained data showed that no statistically significant differences in counts of E. coli and other coliforms on ID and MP media compared with reference methods. However, the evaluation of sensitivity and recovery efficiency of both media showed that the two chromogenic media were more sensitive and significantly more efficient (P = ļ0.05) than reference medium for detecting coliforms in groundwater. However, the identification of 400 typical and atypical colonies isolated from ID and MP media demonstrated a higher specificity when using ID for coliforms and E. coli. In summary, the two chromogenic media evaluated could be used as alternative methods to reference media for detecting and recovering coliform bacteria in groundwater samples. MP agar was more sensitive and efficient than ID agar whereas the latter was more specific and selective.

2020 ◽  
Vol 103 (2) ◽  
pp. 513-522
Author(s):  
Patrick Bird ◽  
Benjamin Bastin ◽  
Nicole Klass ◽  
Erin Crowley ◽  
James Agin ◽  
...  

Abstract Background The 3M™ Petrifilm™ Rapid E. coli/Coliform Count Plate is a selective and differential sample-ready-culture medium designed for the rapid enumeration of Escherichia coli (E. coli) and coliforms in the food and beverage industries. Objective The 3M Petrifilm Rapid E. coli/Coliform Count Plate was compared to the U.S. Food and Drug Administration Bacteriological Analytical Manual (BAM) Chapter 4 Enumeration of Escherichia coli and the Coliform Bacteria, the International Organization of Standards (ISO) 4832:2006 Microbiology of food and animal feeding stuffs — Horizontal method for the enumeration of coliforms—Colony-count technique, and ISO 16649-2:2017 Microbiology of food and animal feeding stuffs—Horizontal method for the enumeration of beta-glucuronidase-positive Escherichia coli—Part 2 Colony-count technique at 44 degrees C using bromo-4-chloro-3- indolyl beta-D-glucuronide methods for the enumeration of E. coli and coliforms in dry dog kibble. Method The candidate method was evaluated using two diluents, Butterfield's phosphate buffered diluent and peptone salt solution, in a paired study design with each reference method in a multi-laboratory collaborative study following the current AOAC Validation Guidelines. Three target contamination levels and an uninoculated control level were evaluated. Results The candidate and reference methods were not statistically different at each contamination level. Reproducibility values obtained during the collaborative study were similar between the candidate and reference methods. Conclusion These results demonstrate that the candidate method is equivalent to the reference methods. Highlight 3M Petrifilm Rapid E. coli/Coliform Count Plate was recommended for Official First Action status for enumeration of E. coli and coliforms in a broad range of foods and environmental surfaces.


2009 ◽  
Vol 89 (2) ◽  
pp. 285-293 ◽  
Author(s):  
S J Bach ◽  
R P Johnson ◽  
K. Stanford ◽  
T A McAllister

Bacteriophage biocontrol has potential as a means of mitigating the prevalence of Escherichia coli O157:H7 in ruminants. The efficacy of oral administration of bacteriophages for reducing fecal shedding of E. coli O157:H7 by sheep was evaluated using 20 Canadian Arcott rams (50.0 ± 3.0) housed in four rooms (n = 5) in a contained facility. The rams had ad libitum access to drinking water and a pelleted barley-based total mixed ration, delivered once daily. Experimental treatments consisted of administration of E. coli O157:H7 (O157), E. coli O157:H7+bacteriophages (O157+phage), bacteriophages (phage), and control (CON). Oral inoculation of the rams with 109 CFU of a mixture of four nalidixic acid-resistant strains of E. coli O157:H7 was performed on day 0. A mixture of 1010 PFU of bacteriophages P5, P8 and P11 was administered on days -2, -1, 0, 6 and 7. Fecal samples collected on 14 occasions over 21 d were analyzed for E. coli O157:H7, total E. coli, total coliforms and bacteriophages. Sheep in treatment O157+phage shed fewer (P < 0.05) E. coli O157:H7 than did sheep in treatment O157. Populations of total coliforms and total E. coli were similar (P < 0.05) among treatments, implying that bacteriophage lysis of non-target E. coli and coliform bacteria in the gastrointestinal tract did not occur. Bacteriophage numbers declined rapidly over 21 d, which likely reduced the chance of collision between bacteria and bacteriophage. Oral administration of bacteriophages reduced shedding of E. coli O157:H7 by sheep, but a delivery system that would protect bacteriophages during passage through the intestine may increase the effectiveness of this strategy as well as allow phage to be administered in the feed.Key words: Escherichia coli O157:H7, bacteriophage, sheep, environment, coliforms


1995 ◽  
Vol 58 (1) ◽  
pp. 13-18 ◽  
Author(s):  
ERROL V. RAGHUBEER ◽  
JIM S. KE ◽  
MICHAEL L. CAMPBELL ◽  
RICHARD S. MEYER

Commercial mayonnaise and refrigerated ranch salad dressing were inoculated at two levels with two strains of Escherichia coli O157:H7, a non-pathogenic E. coli, and the non-fecal coliform Enterobacter aerogenes. Results showed that at the high inoculation level (&gt;106 colony forming units [CFU]/g) in mayonnaise stored at room temperature (ca. 22°C) both strains of O157:H7 were undetected at 96 h. At the high inoculation level, all strains of coliform bacteria tested survived longer in salad dressing stored at 4°C than in mayonnaise stored at 22°C. The O157:H7 strains were still present at low levels after 17 days. The survival time in the low-level inoculum (104CFU/g) study decreased, but the survival pattern in the two products was similar to that observed in the high-level inoculum study. Slight differences in survival among strains were observed. The greater antimicrobial effect of mayonnaise may be attributable to differences in pH, water activity (aw), nutrients, storage temperature, and the presence of lysozyme in the whole eggs used in the production of commercial mayonnaise. Coliform bacteria survived longer in refrigerated salad dressing than in mayonnaise particularly at the high-level inoculum. Both mayonnaise (pH 3.91) and salad dressing (pH 4.51) did not support the growth of any of the microorganisms even though survival was observed.


Author(s):  
J.G.K. Kangumba ◽  
E.H. Venter ◽  
J.A.W. Coetzer

Conventional methods of ensuring the safety and soundness of cows' milk for human consumption, such as pasteurisation, are not always practical in poor socioeconomic conditions or in rural communities that lack modern amenities. Activation of lactoperoxidase (LP) system and souring of milk were investigated as potential alternative methods to sustain the safety of milk by inhibiting certain microorganisms with known pathogenic potential. The activation of the LP-system inhibited the growth of Staphylococcus aureus and Escherichia coli by the order of 2 log values. The inhibition of Brucella abortus was negligible. The replication of Coxiella burnetti in milk was not disturbed even after 17 h of LP-system activation at 20 oC, but the outcome of the LP-system treatment on Mycobacterium bovis could not be determined as the conventional culturing technique used to grow this organism did not allow full recovery. Souring inhibited the growth of S. aureus and E. coli also by the order of 2 log values. From the results obtained in this investigation are concluded that the activation of the LP-system and souring can be used to inhibit the growth of S. aureus and E. coli in cows' milk, thereby increasing its safety.


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


2015 ◽  
Vol 16 (1) ◽  
pp. 31
Author(s):  
Kusdianawati Kusdianawati ◽  
Apon Zaenal Mustopa ◽  
Suharsono Suharsono ◽  
Bugi Ratno Budiarto ◽  
Fatimah Fatimah ◽  
...  

Plantaricin is one of bacteriocins that have the potential to be used as food preservative. Plantaricin is safe for human consumption because it can be easily degraded by proteolytic enzymes. The objective of this study was to express and purify recombinant pre-mature peptide of plantaricin F from <em>Lactobacillus plantarum</em> S34 in <em>Escherichia coli</em>. Plantaricin gene-specific primer was used to obtain pln F structural gene amplicon from L. <em>plantarum</em> S34. This amplicon was cloned in pET32a vector and expressed in E. coli BL21 (DE3) pLysS. Pre-mature plantaricin F peptide was expressed as Histagged-fusion protein and separated by Co2+-chelating affinity chromatography. L. <em>plantarum</em> S34-derived pre-mature plantaricin F peptide fused with thioredoxin-(His)6tag had successfully been expressed in E. <em>coli</em> BL21 (DE3) pLysS using pET32a as an expression vector. The fused recombinant pln F as pre-mature state expressed had a molecular mass of +24 kDa, meanwhile the fused recombinant that contained only the leader peptide of pln F appeared as +20 kDa based on SDS-PAGE separations. The optimal production of fused recombinant pln F as soluble fraction was obtained when culture condition was added with 0.5 mM of IPTG and incubated at 22°C for 5 hours (OD~1). Furthermore, the expression of fused recombinant pln F as its pre-mature peptide pointed out that the pln F’s leader peptide could be proteolytically cleaved by a system in heterologous cells. Overall, heterologous pln F production as pre-mature peptide fused with thioredoxin-(His)6tag had been well established. From this research, we expect plantaricin F can be expressed and purified in E. coli.


2001 ◽  
Vol 64 (11) ◽  
pp. 1756-1760 ◽  
Author(s):  
RAYMOND G. McGUIRE ◽  
ROBERT D. HAGENMAIER

Survival of the coliform bacteria Enterobacter aerogenes and Escherichia coli was monitored in a neutral carboxyme-thylcellulose formulation and in shellac formulations with various pH and concentrations of ethanol and the preservative paraben; populations were subsequently measured from the surface of citrus fruit coated with these formulations. Numbers of the two bacteria increased over 24 h from 106 CFU/ml to approximately 108 CFU/ml in the carboxymethylcellulose solution, but over this time numbers remained little changed in the neutral solution of shellac. The Enterobacter was more tolerant of alcohol over a 3-h period; although its numbers in a shellac solution with 10% ethanol dropped from more than 106 CFU/ml to just over 103 CFU/ml, E. coli and a third species, Klebsiella pneumoniae, declined toward the limit of detection (5 CFU/ml) during this time. The addition of morpholine to increase the formulation pH to 9.0 caused numbers of bacteria to plummet to an undetectable level within 30 to 60 min. On Ruby Red grapefruit and Valencia oranges in storage at 13°C numbers of E. aerogenes and E. coli declined over 2 weeks from 105 CFU/cm2 to less than 2.5 × 101, but most of the loss in numbers occurred within 1 day. Numbers remained significantly less on shellacked fruit compared with those applied in the carboxymethylcellulose coating, and a shellac coating prepared from a pH 9 solution was more toxic to these species than one in which 12% ethanol had been added to the neutral formulation. The addition of the preservative paraben in the basic shellac was further inhibitory.


2017 ◽  
Vol 145 (15) ◽  
pp. 3168-3179 ◽  
Author(s):  
M. K. HENRY ◽  
S. C. TONGUE ◽  
J. EVANS ◽  
C. WEBSTER ◽  
I. J. McKENDRICK ◽  
...  

SUMMARYEscherichia coliO157 are zoonotic bacteria for which cattle are an important reservoir. Prevalence estimates forE. coliO157 in British cattle for human consumption are over 10 years old. A new baseline is needed to inform current human health risk. The BritishE. coliO157 in Cattle Study (BECS) ran between September 2014 and November 2015 on 270 farms across Scotland and England & Wales. This is the first study to be conducted contemporaneously across Great Britain, thus enabling comparison between Scotland and England & Wales. Herd-level prevalence estimates forE. coliO157 did not differ significantly for Scotland (0·236, 95% CI 0·166–0·325) and England & Wales (0·213, 95% CI 0·156–0·283) (P= 0·65). The majority of isolates were verocytotoxin positive. A higher proportion of samples from Scotland were in the super-shedder category, though there was no difference between the surveys in the likelihood of a positive farm having at least one super-shedder sample.E. coliO157 continues to be common in British beef cattle, reaffirming public health policy that contact with cattle and their environments is a potential infection source.


2002 ◽  
Vol 34 (3) ◽  
pp. 227-231 ◽  
Author(s):  
F.M. Schets ◽  
P.J. Nobel ◽  
S. Strating ◽  
K.A. Mooijman ◽  
G.B. Engels ◽  
...  

2006 ◽  
Vol 89 (1) ◽  
pp. 100-114 ◽  
Author(s):  
Hidemasa Kodaka ◽  
Shingo Mizuochi ◽  
Hajime Teramura ◽  
Tadanobu Nirazuka ◽  
David Goins ◽  
...  

Abstract Compact Dry E. coli/Coliform Count (EC) is a ready-to-use test method for the enumeration of Escherichia coli and coliform bacteria in food. The plates are presterilized and contain culture medium and a cold water-soluble gelling agent. The medium should be rehydrated with 1 mL diluted sample inoculated onto the center of the self-diffusible medium, allowing the solution to diffuse by capillary action. The plate can be incubated at 35C for 2024 h and the colonies counted without any further working steps. The Compact Dry EC medium plates were validated as an analysis tool for determining colony-forming units (CFU) of E. coli and coliform bacteria from a variety of raw meats using 5 different types of raw meats. The performance tests were conducted at 35C. In all studies performed, no apparent differences were observed between the Compact Dry ECmethod and theAOAC Official Method 966.24 results. For the accuracy claim (n = 75), a correlation factor of r2 = 0.93 (E. coli) and r2 = 0.93 (coliform bacteria) could be assigned, as stated in the application for Performance-Tested MethodSM.


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