An Improved Membrane Filtration Method for Enumeration of Faecal Coliforms and E. Coli by a Fluorogenic β-Glucuronidase Assay

1993 ◽  
Vol 27 (3-4) ◽  
pp. 267-270 ◽  
Author(s):  
M. T. Augoustinos ◽  
N. A. Grabow ◽  
B. Genthe ◽  
R. Kfir

A fluorogenic β-glucuronidase assay comprising membrane filtration followed by selective enumeration on m-FC agar at 44.5°C and further confirmation using tlie 4-metliylumbelliferyl-β-D-glucuronide (MUG) containing medium was evaluated for the detection of Escherichia coli in water. A total of 200 typical blue and non-typical blue colonies were isolated from sea and fresh water samples using initial selective enumeration on m-FC agar. Pure cultures of the selected colonies were further tested using the MUG assay and identified using the API 20E method. Of the colonies tested which were shown to be positive using the MUG assay 99.4% were Escherichia coli. The results of this study indicate the combination of the m-FC method followed by the MUG assay to be highly efficient for the selection and confirmation of E. coli from a wide range of environmental waters.

1998 ◽  
Vol 61 (4) ◽  
pp. 402-408 ◽  
Author(s):  
CHRISTINE A. POWER ◽  
SCOTT A. MCEWEN ◽  
ROGER P. JOHNSON ◽  
MOHAMED M. SHOUKRI ◽  
KRIS RAHN ◽  
...  

The Petrifilm™ HEC test (3M Canada Inc., London, Ontario), a quantitative microbiological test for Escherichia coli O157:H7, was evaluated for its performance as a beef-carcass monitoring test. Test repeatability and agreement with an E. coli O157:H7 detection method using a hydrophobic grid membrane filter (HGMF) overlaid onto cefixime–tellurite–sorbitol MacConkey agar (CT-SMAC) followed by a latex agglutination test for the O157 antigen were determined by using pure cultures of E. coli O157:H7, beef samples experimentally contaminated with bovine feces containing E. coli O157:H7, and naturally contaminated beef carcasses of unknown E. coli O157:H7 status from a local abattoir. The Petrifilm™ HEC test showed excellent repeatability and excellent agreement with the HGMF–CT-SMAC method when test samples were obtained from pure cultures and experimentally contaminated meat. All 125 naturally contaminated beef carcasses surveyed were negative for E. coli O157:H7 with both microbial methods. The Petrifilm™ HEC test, however, demonstrated a significantly lower proportion of cross-reactive organisms (false-positive reactions) than the HGMF–CT-SMAC method. Given the performance of this test coupled with its ease of use and compact size, it shows considerable promise for carcass testing where abattoir laboratory facilities are limited and as a substitute for more complex laboratory testing methods used in established laboratories.


1991 ◽  
Vol 106 (2) ◽  
pp. 345-354 ◽  
Author(s):  
W. H. S. Cheung ◽  
D. K. K. Ha ◽  
K. Y. Yeung ◽  
R. P. S. Hung

SUMMARYThe standard membrane filtration method of the UK has been modified in order to improve its specificity for enumeratingEscherichia coliin the subtropical waters of Hong Kong. This involves incorporating into the membrane lauryl sulphate (mLS) method either anin situurease test (the mLS-UA method), or anin situβ-glucuronidase test (the mLS-GUD method). The false-positive errors of the mLS-UA and mLS-GUD methods are low, ranging from 3–5%. A comparison between the membrane filtration (mLS-UA) method and the multiple tube technique in testingE. coliin subtropical beach-waters has demonstrated that the former can give much more precise counts, and is the method of choice for such a purpose. The mLS-GUD method, for which automated counting ofE. colicolonies is possible, is a good alternative to mLS-UA in routine enumeration of this bacterial indicator in environmental waters.


1980 ◽  
Vol 85 (2) ◽  
pp. 181-191 ◽  
Author(s):  

SUMMARYIn a multi-laboratory trial with the mombrane filtration technique, three surfactants – Teepol 610 (T610), Tergitol 7 (T7) and sodium lauryl sulphate (LS) – were compared in media for the enumeration of coliform organisms and Escherichia coli in water. A total of 170 samples of water (87 raw and 92 marginally chlorinated) were examined for colony counts of coliform organisms, and 185 water samples (94 raw and 91 marginally chlorinated) for E. coli. Slight differences in the confirmed colony counts between the three media were noted, but few of these were observed consistently in every laboratory. In most laboratories, T7 gave slightly higher counts of E. coli than LS with chlorinated waters; a higher incidence of false-positive results for E. coli at 44 °C was also noted with T7. As there were no outstanding differences in the trial, sodium lauryl sulphate, which is chemically defined, cheap and readily available, is therefore recommended for use at a concentration of 0·1% instead of Teepol 610 in the standard medium for the enumeration of coliform organisms and E. coli in water by the membrane filtration technique.


1982 ◽  
Vol 88 (2) ◽  
pp. 265-273 ◽  
Author(s):  
R. C. Wright

SummaryA selective membrane filtration medium is described for use in the rapid assessment of water quality in tropical countries where the incidence of faecal coliforms other than E. coli presents problems in the interpretation of results. The medium gives comparable results to MPN values obtained in the multiple tube dilution test using modified Gray's glutamate medium, and to membrane filtration counts obtained using M-FC broth and membrane-enriched Teepol broth, whilst differentiation of E. coli is enhanced.


2006 ◽  
Vol 69 (7) ◽  
pp. 1508-1513 ◽  
Author(s):  
MUEEN ASLAM ◽  
CARA SERVICE

The objective of this study was to investigate the extent of antimicrobial resistance and to genetically characterize resistant Escherichia coli recovered from a commercial beef packing plant. E. coli isolates were recovered by a hydrophobic grid membrane filtration method by direct plating on SD-39 medium. A total of 284 isolates comprising 71, 36, 55, 52, and 70 isolates from animal hides, washed carcasses, conveyers, beef trimmings, and ground beef, respectively, were analyzed. The susceptibility of E. coli isolates to 15 antimicrobial agents was evaluated with an automated broth microdilution system, and the genetic characterization of these isolates was performed by the random amplified polymorphic DNA (RAPD) method. Of the 284 E. coli isolates, 56% were sensitive to all 15 antimicrobial agents. Resistance to tetracycline, ampicillin, and streptomycin was observed in 38, 9, and 6% of the isolates, respectively. Resistance to one or more antimicrobial agents was observed in 51% of the E. coli isolates recovered from the hides but in only 25% of the E. coli from the washed carcasses. Resistance to one or more antimicrobial agents was observed in 49, 50, and 37% of the isolates recovered from conveyers, beef trimmings, and ground beef, respectively. The RAPD pattern data showed that the majority of resistant E. coli isolates were genetically diverse. Only a few RAPD types of resistant strains were shared among various sample sources. The results of this study suggest that antimicrobial-resistant E. coli isolates were prevalent during all stages of commercial beef processing and that considerably higher numbers of resistant E. coli were present on conveyers, beef trimmings, and ground beef than on dressed carcasses. This stresses the need for improving hygienic conditions during all stages of commercial beef processing and meatpacking to avoid the risks of transfer of antimicrobial-resistant bacteria to humans.


2018 ◽  
Vol 46 (1) ◽  
pp. 6
Author(s):  
Carolina Maciel Malgarin ◽  
Karine Ludwig Takeuti ◽  
Anne Caroline de Lara ◽  
David Emilio Santos Neves de Barcellos

Background: One of the most frequent health problems in the swine industry is the post-weaning diarrhea in nursery pigs, which leads to significant losses due to weight loss, dehydration, cost of medication and mortality. Escherichia coli  (E. coli) is one of the main bacterial agents of the post-weaning diarrhea. To investigate the possibility of enterotoxigenic  E. coli (ETEC) transmission through drinking water to nursery piglets, the objective of this study was to isolate, characterize by virulence factors, and compare the antimicrobial resistance profiles of E. coli from drinking water samples in nurseries and from rectal swabs of their piglets presenting post-weaning colibacillosis.Materials, Methods & Results: Fifteen rectal swabs from diarrheic piglets in their first three weeks after weaning and one water sample were collected from each of ten nurseries located in Rio Grande do Sul State, south of Brazil. After enrichment with a commercial broth medium, water samples were cultured in blood agar, as well as the rectal swab samples, and the characteristic colonies were identified by standard biochemical analysis. Following isolation and identification of E. coli, the colonies from water samples and their corresponding piglets’ samples were characterized by multiplex PCR in order to determine specific ETEC fimbria and toxin genes. Finally, all E. coli isolates were submitted to antimicrobial susceptibility testing. Virulence factors and antimicrobial sensitivity could then be compared between water and piglets’ samples. The difference in the antimicrobial resistance frequency for each of the sample groups were compared using the multi comparison test. E. coli was isolated in four out of the ten water samples, although none of the water samples presented ETEC virulence factors. From 60 rectal swab samples (15 from each of the four positive farms with E. coli isolated from water samples), 21 E. coli were isolated and seven demonstrated characteristic ETEC virulence factors. The fimbriae exhibited in higher frequency were F18 (62.5%) and F4 (25%) and the toxins were STb (100%) and STaP (75%). E. coli isolated from water samples presented higher resistance to the antimicrobials apramycin, florfenicol, lincomycin, lincomycin+spectinomycin, oxytetracycline, and sulfamethoxazole+trimethoprim; it did not present resistance to colistin and fosfomycin. The seven ETEC from rectal swab samples presented a higher resistance to lincomycin, and lower resistance frequency to fosfomycin. The other 14 E. coli non-ETEC from rectal swab samples presented a higher resistance to florfenicol and no resistance to colistin.Discussion: Enterotoxigenic E. coli is an important agent causing post-weaning colibacillosis, although, differently from other studies, this experiment did not find the agent in most of the sampled animals. In contrast to other authors, ETEC was not found in water, as the development of its virulence factors may depend on conditions presented exclusively in the animal. By the results we can conclude that, although E. coli was isolated from the drinking water, it was not a significant mechanism for nursery piglets’ infection with ETEC in this experiment. The samples analyzed presented a wide range of resistance to different antimicrobials, including multi-resistance. In some cases, E. coli found in water presented different antimicrobial profile from the bacterium found in the rectal swab samples. Enterotoxigenic E. coli was susceptible to fosfomycin and its use may represent a prudent antimicrobial choice to the swine industry.


1991 ◽  
Vol 37 (12) ◽  
pp. 908-911 ◽  
Author(s):  
Lois C. Shadix ◽  
Eugene W. Rice

The new United States Drinking Water Regulations state that water systems must analyze for Escherichia coli or fecal coliforms on any routine or repeat sample that is positive for total coliforms. The proposed methods for the detection of E. coli are based on β-glucuronidase activity, using the fluorogenic substrate 4-methylumbelliferyl β-D-glucuronide (MUG). This study was conducted to determine whether β-glucuronidase negative E. coli were present in significant numbers in environmental waters. Two hundred and forty E. coli cultures were isolated from 12 water samples collected from different environmental sources. β-glucuronidase activity was determined using lauryl tryptose broth with MUG, EC broth with MUG, and the Autoanalysis Colilert (AC) procedure. The isolates were also evaluated by the standard EC broth gas fermentation method for fecal coliforms. The results confirm that assaying for the enzyme β-glucuronidase utilizing the MUG substrate is an accurate method for the detection of E. coli in environmental waters. Key words: Escherichia coli, β-glucuronidase, 4-methylumbelliferyl β-D-glucuronide, water.


2017 ◽  
Vol 80 (10) ◽  
pp. 1670-1675 ◽  
Author(s):  
J. Pérez-Garza ◽  
S. García ◽  
N. Heredia

ABSTRACT Foodhandlers are important sources of contamination in the agricultural environment. This study was conducted (i) to evaluate the activity of antimicrobial soaps against Escherichia coli and Enterococcus faecalis using a hand washing model with soiled hands and (ii) to determine the survival and persistence of these bacteria in rinsates. Sterilized agricultural soil from tomato and pepper farms was inoculated with E. coli or E. faecalis at 103 or 106 CFU/g. Decontaminated hands were placed in contact with contaminated soil for 2 min and were then washed with soaps with or without antimicrobial compounds (citric extracts, chloroxylenol, triclosan, or chlorhexidine gluconate). As the control, hands were washed with sterile distilled water. The levels of bacteria remaining on the hands and recovered from the rinsates were determined using a membrane filtration method and selective media. Antimicrobial soaps removed levels of E. coli similar to those removed by distilled water and nonantimicrobial soap on hands contaminated with E. coli at 103 CFU/g. However, when hands were contaminated with E. coli at 106 CFU/g, more E. coli was removed with the chlorhexidine gluconate soap. When hands were contaminated with E. faecalis at 103 CFU/g, bacteria were removed more effectively with soaps containing chloroxylenol or chlorhexidine gluconate. When hands were contaminated with E. faecalis at 106 CFU/g, all of the antimicrobial soaps were more effective for removing the bacteria than were distilled water and nonantimicrobial soap. E. coli grew in all of the hand washing rinsates except that containing triclosan, whereas E. faecalis from the 106 CFU/g treatments grew in rinsates containing chlorhexidine gluconate and in the distilled water rinsates. Washing with antimicrobial soap was more effective for reducing bacteria on soiled hands than was washing with water or nonantimicrobial soap. However, persistence or growth of bacteria in these rinsates poses health risks.


2022 ◽  
Author(s):  
ADEWALE OLUWASOGO OLALEMI ◽  
OLUWABUSAYOMI MARY OLUYEMI ◽  
MICHAEL TOSIN BAYODE

Abstract Background This study was carried out to monitor the levels of faecal pollution markers in catfish (Clarias gariepinus) and their growing waters in selected earthen and concrete ponds. Water and catfish samples were collected weekly in the months of February, March, April, May, June and July, 2019. The concentrations of enteric bacteria in the water and catfish samples were determined using membrane filtration and pour plate methods, respectively. The rate of bioaccumulation of faecal indicator bacteria was obtained by dividing the log concentration of each organism in catfish by the corresponding log concentration in the growing waters. Result The concentration of faecal coliforms in catfish samples from concrete and earthen ponds ranged from 1.41 to 2.28 log10 CFU/100 ml and 1.3 to 2.47 log10 CFU/100 ml respectively and in growing waters from the concrete and earthen ponds; 1.43 to 2.41 log10 CFU/100 ml and 1.50 to 2.80 log10 CFU/100 ml respectively. Faecal coliforms exhibited positive relationships with alkalinity in water samples from the earthen (r = 0.61) and concrete ponds (r = 0.62). Salmonella and faecal coliforms had the highest and least bioaccumulation in catfish raised in earthen pond, respectively, whereas Salmonella and enterococci had the highest and least bioaccumulation in catfish raised in concrete pond, respectively. Faecal coliforms and E. coli had the highest and least counts in water samples from the earthen pond during the dry and wet months, Salmonella and E. coli had the highest and least counts in water samples from the concrete pond during the dry and wet months. Conclusion There were high levels of bacterial faecal pollution markers in water and C. gariepinus from the earthen and concrete ponds. Physicochemical characteristics of the water and seasonality played major roles in the rate of bioaccumulation of the faecal pollution markers in C. gariepinus raised in the earthen and concrete ponds.


Gut Pathogens ◽  
2019 ◽  
Vol 11 (1) ◽  
Author(s):  
Zahid Hayat Mahmud ◽  
Md Shafiqul Islam ◽  
Khan Mohammad Imran ◽  
Syed Adnan Ibna Hakim ◽  
Martin Worth ◽  
...  

Abstract Background Safe water is essential for life but unsafe for human consumption if it is contaminated with pathogenic microorganisms. An acceptable quality of water supply (adequate, safe and accessible) must be ensured to all human beings for a healthy life. Methods We collected and analyzed a total of 12,650 drinking water samples, for the presence of Escherichia coli and faecal coliforms, from a large habitation of the displaced Rohingya population comprising of about 1.16 million people living within 4 km2. Results We found that 28% (n = 893) water samples derived from tubewells were contaminated with faecal coliforms and 10.5% (n = 333) were contaminated with E. coli; also, 73.96% (n = 4644) samples from stored household sources (at point of use—POU) were found contaminated with faecal coliforms while 34.7% (n = 2179) were contaminated with E. coli. It was observed that a higher percentage of POU samples fall in the highest risk category than that of their corresponding sources. Conclusions From our findings, it appears that secondary contamination could be a function of very high population density and could possibly occur during collection, transportation, and storage of water due to lack of knowledge of personal and domestic hygiene. Hence, awareness campaign is necessary, and the contaminated sources should be replaced. Further, the POU water should be treated by a suitable method.


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