Evaluation of m-T7 agar as a fecal coliform medium

m-T7 agar, designed to improve recoveries of injured total coliforms, was evaluated for its effectiveness as a fecal coliform medium. The time and temperature of preincubation were found to be crucial to the optimal recovery of fetal coliforms. Isolation rates for fecal coliforms on m-T7 agar from sewage effluents were the highest when plates were preincubated at 37 degrees C for 8 h before transfer to 44.5 degrees C for 12 h. The medium was found to produce consistently higher fecal coliform counts than all the other methods tested. Recoveries were 3.1 times greater than the standard m-FC method and 1.7 times greater than the two-layer enrichment, temperature acclimation procedure. Verification rates for fecal coliforms isolated on m-T7 agar averaged 89.0%, whereas verification rates for m-FC agar averaged only 82.8%. Both media isolated similar fecal coliform populations. The advantages of a single medium, highly effective for the isolation of both total and fecal coliforms, are discussed.     

Comparison of fecal coliform agar and violet red bile lactose agar for fecal coliform enumeration in foods

A 24-h direct plating method for fecal coliform enumeration with a resuscitation step (preincubation for 2 h at 37 +/- 1 degrees C and transfer to 44 +/- 1 degrees C for 22 h) using fecal coliform agar (FCA) was compared with the 24-h standardized violet red bile lactose agar (VRBL) method. FCA and VRBL have equivalent specificities and sensitivities, except for lactose-positive non-fecal coliforms such as Hafnia alvei, which could form typical colonies on FCA and VRBL. Recovery of cold-stressed Escherichia coli in mashed potatoes on FCA was about 1 log unit lower than that with VRBL. When the FCA method was compared with standard VRBL for enumeration of fecal coliforms, based on counting carried out on 170 different food samples, results were not significantly different (P > 0.05). Based on 203 typical identified colonies selected as found on VRBL and FCA, the latter medium appears to allow the enumeration of more true fecal coliforms and has higher performance in certain ways (specificity, sensitivity, and negative and positive predictive values) than VRBL. Most colonies clearly identified on both media were E. coli and H. alvei, a non-fecal coliform. Therefore, the replacement of fecal coliform enumeration by E. coli enumeration to estimate food sanitary quality should be recommended.     

Rapid seven-hour fecal coliform test.

A rapid 7-h fecal coliform (FC) test for the detection of FC in water has been developed. This membrane filter test utilizes a lightly buffered lactose-based medium (m-7-h FC medium) combined with a sensitive pH indicator system. FC colonies appeared yellow against a light purple background after incubation at 41.5 degrees C for 7 to 7.25 h. Comparison of FC test results showed that the mean verified FC count ratio (7-h FC count/24-h FC count) for surface water samples was 1.08. The mean FC count ratio (7-h FC count/24-h FC count) for unchlorinater wastewater ranged from 1.95 to 5.05. Verification of yellow FC colonies from m-7-h FC medium averaged 97%. Data from field tests on Lake Michigan bathing beach water samples showed that unverified 7-h FC counts averaged 96% of the 24-h FC counts. The 7-h FC test was found to be suitable for the examination of surface waters and unchlorinated sewage and could serve as an emergency test for detection of sewage or fecal contamination of potable water.     

Rapid seven-hour fecal coliform test.

A rapid 7-h fecal coliform (FC) test for the detection of FC in water has been developed. This membrane filter test utilizes a lightly buffered lactose-based medium (m-7-h FC medium) combined with a sensitive pH indicator system. FC colonies appeared yellow against a light purple background after incubation at 41.5 degrees C for 7 to 7.25 h. Comparison of FC test results showed that the mean verified FC count ratio (7-h FC count/24-h FC count) for surface water samples was 1.08. The mean FC count ratio (7-h FC count/24-h FC count) for unchlorinater wastewater ranged from 1.95 to 5.05. Verification of yellow FC colonies from m-7-h FC medium averaged 97%. Data from field tests on Lake Michigan bathing beach water samples showed that unverified 7-h FC counts averaged 96% of the 24-h FC counts. The 7-h FC test was found to be suitable for the examination of surface waters and unchlorinated sewage and could serve as an emergency test for detection of sewage or fecal contamination of potable water.     

A-1 medium: alternative technique for fecal coliform organism enumeration in chlorinated wastewaters.

A 24-h most-probable-number technique using A-1 medium for detecting fecal coliforms in chlorinated wastewaters was evaluated. The A-1 medium technique, using 3 h of preincubation at 35 degrees C, gave results statistically equivalent to those obtained with the American Public Health Association Standard Methods two-step most-probable-number technique.     

A new selective and differential agar medium for Escherichia coli and coliform organisms

W right , R.C. 1984. A new selective and differential agar medium for Escherichia coli and coliform organisms. Journal of Applied Bacteriology 56 , 381–388.
An enriched lauryl sulphate-aniline blue agar medium which is selective for Escherichia coli and coliform organisms is described. From faecal samples, the medium gave higher counts of colonies producing acid from lactose than media containing bile salts. From contaminated water and food samples, the medium gave comparable or higher counts of colonies identified as E. coli than standard media. Colonies of E. coli were more readily differentiated from those of other coliform organisms.     

A new selective and differential agar medium for Escherichia coli and coliform organisms

An enriched lauryl sulphate-aniline blue agar medium which is selective for Escherichia coli and coliform organisms is described. From faecal samples, the medium gave higher counts of colonies producing acid from lactose than media containing bile salts. From contaminated water and food samples, the medium gave comparable or higher counts of colonies identified as E. coli than standard media. Colonies of E. coli were more readily differentiated from those of other coliform organisms.     

Microplate fecal coliform method to monitor stream water pollution.

A study has been carried out on the Moselle River by means of a microtechnique based on the most-probable-number method for fecal coliform enumeration. This microtechnique, in which each serial dilution of a sample is inoculated into all 96 wells of a microplate, was compared with the standard membrane filter method. It showed a marked overestimation of about 14% due, probably, to the lack of absolute specificity of the method. The high precision of the microtechnique (13%, in terms of the coefficient of variation for log most probable number) and its relative independence from the influence of bacterial density allowed the use of analysis of variance to investigate the effects of spatial and temporal bacterial heterogeneity on the estimation of coliforms. Variability among replicate samples, subsamples, handling, and analytical errors were considered as the major sources of variation in bacterial titration. Variances associated with individual components of the sampling procedure were isolated, and optimal replications of each step were determined. Temporal variation was shown to be more influential than the other three components (most probable number, subsample, sample to sample), which were approximately equal in effect. However, the incidence of sample-to-sample variability (16%, in terms of the coefficient of variation for log most probable number) caused by spatial heterogeneity of bacterial populations in the Moselle River is shown and emphasized. Consequently, we recommend that replicate samples be taken on each occasion when conducting a sampling program for a stream pollution survey.     

Comparison of four membrane filter methods for fecal coliform enumeration.

Four membrane filter methods fecal coliform enumeration were evaluated and compared in six laboratories based on determination of accuracy, specificity, upper counting limit, and recovery comparability. Recovery accuracy with pure cultures ranged from 89 to 100% for m-FC, mTEC (a procedure developed for thermotolerant Escherichia coli), and m-FC2 methods (the latter incorporating a 2-h, 35 degrees C resuscitation period), but was less than 60% for the MacConkey membrane broth method. These figures dropped by approximately 40 to 55% when the cultures were subjected to temperature (10 degrees C) stress. Close to 800 colonies were verified to determine specificity. False-positive colonies occurred most frequently with the m-FC2 method (18%), whereas false-negative colonies were most common on MacConkey membrane broth (26%). In counting range experiments using a variety of samples, the highest upper counting limit was 130 colonies per filter with the mTEC procedure. Recovery comparisons were based on over 130 samples including raw surface waters, raw sewage, and chlorinated and unchlorinated sewage effluents. In general, recoveries were significantly higher with the m-FC2 and mTEC methods; however, on m-FC2, growth of nontarget background organisms was also higher in most cases. Highest recoveries from chlorinated sewage effluents were obtained by the mTEC method, and the addition of a similar resuscitation period to the m-FC procedure (m-FC2) improved fecal coliform recovery from such samples. The best overall performance for fecal coliform enumeration was obtained with the mTEC method with high recovery and low levels of background colonies, good specificity and accuracy, and a high upper counting limit. This procedure also offers the advantage of enumerating E. coli within 24 h.     

Evaluation of a new chromogenic agar medium for the identification of urinary tract pathogens

This study evaluated the performance of CPS ID2 (bioMérieux) compared to that of conventionally used selective agar for the identification of bacteria responsible for urinary tract infections. This medium detects bacterial enzymes using chromogenic substrates. Two hundred and nineteen samples of urine were tested in order to evaluate new CPS ID2 in comparison to blood agar and MacConkey agar.
According to our results, the CPS ID2 agar is an easy, rapid and sensitive method for the screening of colonies suspected of being Escherichia coli , reducing the number of biochemical tests needed. Also, enterococci and Proteae can be easily detected. Other micro-organisms require further identification. The greatest value of this medium is the accurate identification of polymicrobial cultures.     

Effect of dredge spoil deposition on fecal coliform counts in sediments at a disposal site.

The most-probable-number of fecal coliforms in sediments was monitored at the New London dump site in Long Island Sound during the deposition of dredge spoil from the Thames River. Although the geometric mean for fecal coliforms at five stations in the river was 14,000/100 ml before dredging commenced, the deposition of this material did not increase the incidence of fecal coliforms at 17 spoil stations and 13 control stations in the disposal and surrounding areas. Fecal coliforms appear to occur only in the surface sediment material and are diluted by the subsurface material during the dredging operation. Fecal coliform analyses of bottom waters during high and low tides indicated that the flow of water from the Thames River played a major role in determining the most-probable-number of fecal coliforms in the sediments at the disposal site.     

Incidence of R factors in coliform, fecal coliform, and Salmonella populations of the Red River in Canada.

Coliforms, fecal coliforms, and Salmonella were isolated from the Red River, Manitoba, Canada, and identified. These organisms were then examined for resistance to 12 antibiotics. Some fecal coliforms were resistant to all 12 antibiotics, and 18% of the Salmonella isolates were resistant to one or more antibiotics. A total of 52.9% of the fecal coliforms resistant to three or more antibiotics were able to transfer single or multiple resistance (R) determinants to the Salmonella recipient, and 40.7% could transfer R determinants to the Escherichia coli recipient. Of the resistant Salmonella, 57% transferred one or two determinants to the Salmonella recipient, and 39% transferred one or two determinants to the E. coli recipient. It was calculated that populations of fecal coliforms containing R factors were as high as 1,400 per 100 ml and that an accidental intake of a few milliliters of water could lead to transient or permanent colonization of the digestive tract. Consideration of data on bacteria with R factors should be made in future water quality deliberations and in discharge regulations.     

Effect of dredge spoil deposition on fecal coliform counts in sediments at a disposal site.

The most-probable-number of fecal coliforms in sediments was monitored at the New London dump site in Long Island Sound during the deposition of dredge spoil from the Thames River. Although the geometric mean for fecal coliforms at five stations in the river was 14,000/100 ml before dredging commenced, the deposition of this material did not increase the incidence of fecal coliforms at 17 spoil stations and 13 control stations in the disposal and surrounding areas. Fecal coliforms appear to occur only in the surface sediment material and are diluted by the subsurface material during the dredging operation. Fecal coliform analyses of bottom waters during high and low tides indicated that the flow of water from the Thames River played a major role in determining the most-probable-number of fecal coliforms in the sediments at the disposal site.     

Phosphorus and carbohydrate limitation of fecal coliform and fecal enterococcus within tidal creek sediments

Aquatic sediments can be a significant reservoir of bacterial indicators of fecal contamination at levels higher than the waters above them. Several environmental factors have been identified that can enhance the role of sediments as a reservoir for enteric pathogens, including carbon and/or phosphorus availability. In order to investigate the influence of these and other environmental factors on sediment fecal bacteria populations, sediment samples were collected from a coastal watershed in southeastern North Carolina and analyzed for fecal coliform and fecal enterococcus using a modified membrane filtration technique. Measurements of sediment phosphorus, sediment carbohydrate, and environmental factors were made and relationships with bacteria concentrations were assessed. These observations were accompanied by an experimental laboratory manipulation of phosphorus and carbohydrate and their effects on sediment-associated fecal coliform and enterococcus. Field results suggested that sediment-associated indicator bacteria were not limited by sediment phosphorus or carbohydrate. Experimental results suggested that sediment-associated fecal bacteria were more frequently limited by bioavailable carbohydrate. Sediment phosphorus was limiting for fecal enterococcus only where sediment P was initially low (<31 μg P g⁻¹). A strong positive response by sediment fecal coliform concentrations to recent (24 h) precipitation was evidence that stormwater runoff delivers fecal bacteria loadings that are only partly measurable by conventional water sampling schemes, and by driving sediment and sediment P-loading plays a significant role in enhancing aquatic sediments as reservoirs for fecal microbes.     

Evaluation of a new chromogenic agar medium for isolation and identification of Group B streptococci

AIMS: To evaluate a new chromogenic agar as a screening medium for the isolation of Group B streptococci from high vaginal swabs from pregnant women. METHODS AND RESULTS: The medium was evaluated with 195 high vaginal swabs referred for antenatal screening and compared with blood agar and Granada medium. The new chromogenic medium showed 100% sensitivity for the detection of Group B streptococci, and also showed a positive predictive value of 100%. Granada medium also showed excellent sensitivity and specificity and both media were superior to blood agar. CONCLUSIONS: The new chromogenic medium showed excellent performance for the detection of Group B streptococci from clinical samples. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first chromogenic medium described for the detection of Group B streptococci. The medium offers an effective and convenient alternative to conventional media, currently used in clinical laboratories.     

Improved method of screening for aflatoxin with a coconut agar medium.

Nine isolates of Aspergillus flavus and Aspergillus parasiticus were screened for aflatoxin production on a coconut extract agar medium. Aflatoxin-producing colonies were detected under long-wave UV light (365 nm) by blue fluorescence on the reverse side after 2 to 5 days of growth. Aflatoxin production was verified by chemical analysis. Several types of shredded coconut available in the United States were tested and found to be satisfactory. No additives were required. Various parameters affecting the test were investigated.