Detection Of Coliform Bacteria Research Articles

Detection of Coliform Bacteria in Raw Milk Samples Collected from Industrial Cities of Pakistan

Antibiotics and heavy metals-resistant bacteria in livestock environments can result in economic losses and raise public health and environmental problems. There is a crisis in the world's access to and pipeline for antibiotics. Objective: To screen raw milk samples collected from three different industrial cities Gujranwala, Lahore, and Sheikhupura, situated in the province of Punjab, Pakistan. Methods: In this regard, a total of 26 samples were having 84 coliform strains. Separated coliform colonies were processed for Gram's staining, catalase, indole production, and Simmon's citrate and motility tests. Results: Metal resistance of bacterial strains was also checked and 39.5% and 45.23% of bacteria were found to be resistant to ZnCl2 1% and 0.5%. 69.045% and 77.38% bacteria were found to be resistant to CuSo4 salt solution1% and 0.5%. 17.85% and 27% bacteria were found to be resistant to Na2CrO4 salt solution1% and 0.5% respectively. 80% of bacteria were found to be resistant to Cefuroxime, 26.19% to Cephradine, 84.52% to Aztroeonam 41.67% to Erythromycin, 91.667% to Trimethoprim 89.28% to Lincomycins. Conclusions: The raw milk samples were not only contaminated with coliforms but the bacteria were also resistant to heavy metals and certain antibiotics which might be considered indicative of industrial and anthropogenic pollution. Cephradine, 84.52% to Aztroeonam 41.67% to Erythromycin, 91.667% to Trimethoprim 89.28% to Lincomycins. Conclusions: The raw milk samples were not only contaminated with coliforms but the bacteria were also resistant to heavy metals and certain antibiotics which might be considered indicative of industrial and anthropogenic pollution.

Cefsulodin and Vancomycin: A Supplement for Chromogenic Coliform Agar for Detection of Escherichia coli and Coliform Bacteria from Different Water Sources.

Background microorganism growth on Chromogenic Coliform Agar (CCA) can be challenging. For this reason, a new alternative method with a Cefsulodin/Vancomycin (CV)-supplemented CCA should be developed in this study. CCA supplemented with CV was validated according to ÖNORM EN ISO 16140-4:2021 using water from natural sources in Styria, Austria. Results show that the alternative method using the supplemented CCA has similar values in relation to sensitivity (82.2%), specificity (98.6%) and higher selectivity (59%) compared to the reference method. Repeatability and reproducibility were acceptable for the alternative method and showed similar results with the reference method. The alternative method shows a very low false positive rate and a low false negative rate paired with good performance regarding the inclusion study. The exclusion study shows the advantage of our method by suppressing background microorganisms and facilitating the process of enumeration of Escherichia coli and other coliform bacteria on CCA plates. Aeromonas hydrophila and Pseudomonas aeruginosa growth was inhibited using the supplement. To conclude, the coliform CV selective supplement combined with CCA is an appropriate tool for coliform bacteria detection in water samples.

Validation of the Soleris® Coliform Method for Detection of Coliform Bacteria in Dried Cannabis Flower: AOAC Performance Tested MethodSM 010302.

The Soleris® Coliform Vial is a growth-based, automated method for detection of coliform bacteria in foods and other consumer products such as nutraceuticals and cosmetics. The method was granted AOAC Performance Tested MethodSM certification for select foods after successful completion of a validation study. The objective of the current study was to validate the Soleris coliform method for use with dried cannabis flower (>0.3% delta 9-tetrahydrocannabinol). A comparative matrix study was performed in which naturally contaminated dried cannabis flower was tested with the Soleris coliform method and with the U.S. Food and Drug Administration Bacteriological Analytical Manual solid medium method. Multiple lots of dried cannabis flower were obtained, pre-screened for coliforms, and blended to produce test materials at four different contamination levels ranging from 4.5 to 1600 CFU/g. Each material was tested at three different Soleris detection threshold levels determined by the dilution used to inoculate the Soleris vials. Probability of detection analysis was performed to determine if differences in the number of positive results obtained with the two methods were significant. For all four dried cannabis flower materials, at all three Soleris test thresholds, there were no significant differences in the number of positive results obtained with the Soleris and cultural plating methods as determined by probability of detection analysis at P < 0.05. The Soleris coliform test is an accurate method for detection of coliform bacteria in dried cannabis flower. The Soleris coliform method provides cannabis industry QC personnel with an effective method for analysis of dried cannabis flower and produces results in 18-24 h.

Microbial Contamination Survey of Environmental Fresh and Saltwater Resources of Upolu Island, Samoa

Analysis of microbe diversity in freshwater resources and nearshore seawater samples of Upolu Island was performed to investigate the distribution of harmful bacteria. For this, 124 samples were collected from 23 river systems, two volcanic lakes, and 45 locations inside and outside the barrier reef of Upolu Island, Samoa. Physicochemical parameters for general water quality, detection of coliform bacteria and 16S rRNA amplicon sequencing were performed on all samples. Fecal indicator bacteria (FIB) testing indicated a wide distribution of coliform bacteria in all sampled freshwater sites with evidence of fecal coliform in most locations. Importantly, evidence of coliform bacteria was found in most seawater samples inside and along the reef, apart from those samples taken ~20 km offshore. Illumina 16S rRNA amplicon sequencing of the V4 hypervariable region confirmed the presence of various types of harmful bacterial species, namely from the Enterobacteriaceae, Enterococcae, Streptococcaceae, and Vibrionacea families. By combining the sensitivity of FIB testing and next-generation sequencing, we were able to show the extent of potential contaminations in fresh and seawater samples and simultaneously identify the potential pathogenic bacterial genera present. The wide distribution of potential harmful bacteria from river runoff or direct sewage dumping has an impact on human health, leading to many skin and intestinal diseases, and is potentially detrimental to coral reef community health.

Detection of coliform bacteria, total plate count and pH value in chicken eggs from Maros traditional market

Indonesian consumers generally buy chicken eggs in traditional markets. Mostly, the traditional markets were less clean places, making it easier for bacterial contamination to the commodity. The purpose of this study was to determine the presence of Coliform bacteria, Total Plate Count (TPC) and pH value of chicken eggs from the traders in traditional markets in Maros regency. Chicken egg samples were obtained from 3 traditional markets in Maros Regency, each from 3 sellers and 3 replications. The number of coliform bacteria was determined using Brilliance Agar media while the TPC was determined using Plate Count Agar. The pH value was measured using a pH meter. The research method used was a pour plate to count Coliform and TPC bacteria then analyzed descriptively and presented in tabular form. The pH value was analyzed using analysis of variance and further test of the least significant difference. Based on SNI-3926-2008 regarding the microbiological quality requirements of fresh eggs, all egg samples met the Coliform and TPC bacteria standard, with the number of 1.1×101 and 2.3×104, respectively. The pH of eggs was still in the normal range with an average of 7.1 although the type of market affects the pH of the eggs. All egg samples studied based on Coliform bacteria, TPC and pH were categorized as safe for consumption.

The single-tube quantitative rapid detection of coliform bacteria based on enzyme-specific technology.

To detect coliform bacteria (CB) with simplicity and portability, a novel, single-tube quantitative rapid detection method was developed based on CB-specific enzymatic technology. This technology involved a simple multi-channel spectrometer which could detect 7 cfu mL-1 CB specifically, and the approach was applied to various food and water samples.

Development of Fluorescence In Situ Hybridization as a Rapid, Accurate Method for Detecting Coliforms in Water Samples.

Coliform bacteria are indicators of water quality; however, most detection methods for coliform bacteria are time-consuming and nonspecific. Here, we developed a fluorescence in situ hybridization (FISH) approach to detect four types of coliform bacteria, including Escherichia coli, Klebsiella pneumoniae, Enterobacter aerogenes, and Citrobacter freundii, simultaneously in water samples using specific probes for 16S rRNA. This FISH method was applied to detect coliform bacteria in simulated water and domestic wastewater samples and compared with traditional detection methods (e.g., plate counting, multiple-tube fermentation (MTF) technique, and membrane filter (MF) technique). Optimal FISH conditions for detecting the four types of coliforms were found to be fixation in 3% paraformaldehyde at 4 °C for 2 h and hybridization at 50 °C for 1.5 h. By comparing FISH with plate counting, MTF, MF, and a commercial detection kit, we found that FISH had the shortest detection time and highest accuracy for the identification of coliform bacteria in simulated water and domestic wastewater samples. Moreover, the developed method could simultaneously detect individual species and concentrations of coliform bacteria. Overall, our findings indicated that FISH could be used as a rapid, accurate biosensor system for simultaneously detecting four types of coliform bacteria to ensure water safety.

Analisis Total Coliform di Air dan Sedimen di Pesisir Jabon, Sidoarjo Jawa Timur

Penelitian ini bertujuan : (1) menghitung Total Coliform air dan sedimen; (2)membandingkan data parameter fisika,kimia dan Total Coliform dengan Keputusan Menteri Lingkungan Hidup No.51 Tahun 2004 Lampiran 2 dan 3; (3) mengetahui perbedaan Total Coliform air dan sedimen serta (4) mengelompokkan kesamaan data berdasarkan parameter lingkungan dan Total Coliform menggunakan clusterring analysis. Metode penghitungan Total Coliform menggunakan metode MPN dan uji coliform menggunakan metode tabung fermentasi ganda. Berdasarkan uji deteksi bakteri coliform, didapatkan Total Coliform air sebesar 21-70 MPN/100ml dan Total Coliform sedimen 110-9.000 MPN/100 ml. Hal ini menunjukkan bahwa Pesisir Jabon mengalami penurunan kualitas lingkungan perairan laut berdasarkan bakteri coliform.

Predicting coliform presence in private wells as a function of well characteristics, parcel size and leachfield soil rating

Public water systems must be tested frequently for coliform bacteria to determine whether other pathogens may be present, yet no testing or disinfection is required for private wells. In this paper, we identify whether well age, type of well, well depth, parcel size, and soil ratings for a leachfield can predict the probability of detecting coliform bacteria in private wells using a multivariate logistic regression model. Samples from 1163 wells were analyzed for the presence of coliform bacteria between October 2017 and October 2019 across Gaston County, North Carolina, USA. The maximum well age was 30 years, and bored wells (median age = 24 years) were older than drilled wells (median age = 19 years). Bored wells were shallower (mean depth = 18 m) compared to drilled wells (mean depth = 79 m). We found coliform bacteria in 329 samples, including 290 of 1091 drilled wells and 39 of 72 bored wells. The model results showed bored wells were 4.76 times more likely to contain bacteria compared to drilled wells. We found that the likelihood of coliform bacteria significantly increased with well age, suggesting that those constructed before well standards were enforced in 1989 may be at a higher risk. We found no significant association between poorly rated soils for a leachfield, well depth, parcel size and the likelihood of having coliform in wells. These findings can be leveraged to determine areas of concern to encourage well users to take action to reduce their risk of drinking possible pathogens in well water.

ANGKA PALING MUNGKIN DAN DETEKSI COLIFORM PADA SAMPEL LALAPAN DAUN KEMANGI (Ocimum bacilicum) DI KOTA PONTIANAK

Ocimum basilicum is one type of vegetables that can be consumed without having to be processed. Food consumed must have high nutritional value, clean and free of microbes. Coliform is one of the bacteria that can cause disease in humans. This study aims to determine the value of MPN on samples Ocimum basilicum leaves from warung lamongan in Pontianak and determine the existence of coliform bacteria on Ocimum basilicum leaf samples from warung lamongan in Pontianak. Detection of coliform bacteria using the MPN method. Detection of E. coli was carried out using EMBA media. The results showed that all samples had interval values between 210 MPN/g to &gt; 1100 MPN/g. It showed that all samples excced the threshold determined by BPOM (2016), which was the maximum limit of coliform bacteria contamination in ready-toconsume vegetables was &lt; 3 APM/g. The results of the identification of the bacterial genus coliform grouphad the members of genera Escherichia and Citrobacter

Evaluation of Microbiological and Free-Living Protozoa Contamination in Dental Unit Waterlines.

Studies conducted over the last 40 years have demonstrated that the water output from dental unit waterlines (DUWLs) is often contaminated with high densities of microorganisms. It has been monitored the microbiological quality of the water in 30 public dental facilities in northern Italy in order to assess the health risk for patients and dental staff. In each facility, samples of water both from taps and from DUWLs were analyzed in order to evaluate heterotrophic plate counts (HPCs) at 22 °C and 36 °C, and to detect coliform bacteria, Pseudomonas aeruginosa, Legionella pneumophila and amoebae. In 100% of the samples taken from the DUWLs, the concentration of HPCs was above the threshold as determined by the Ministère de la Santé et des Solidarités (2007). The concentration of P. aeruginosa was greater than the indicated threshold in 16.67% of the hand-pieces analyzed. A total of 78.33% of samples were contaminated by L. pneumophila, while in the samples taken from the DUWLs alone, this percentage rose to 86.67%. Amoebae were detected in 60% of the samples taken from hand-pieces; all belonging to the species V. vermiformis. This study documented the presence of various microorganisms, including Legionella spp., at considerably higher concentrations in water samples from DUWLs than in samples of tap water in the same facilities, confirming the role of the internal DUWLs in increasing microbial contamination, especially in the absence of proper management of waterborne health risks.

Rapid detection of coliform bacteria using a lateral flow test strip assay

Coliform bacteria in foods are enumerated at food processing plants and are used as sanitary and quality indicators. To detect coliform bacteria rapidly, seven Lateral Flow Test Strips (LFTSs) that can detect the genera Aeromonas, Citrobacter, Enterobacter, Hafnia, Klebsiella/Raoultella, Pantoea and Serratia were developed. For 55 tested food isolates, the detection rate of each individual LFTS assay was only 38% to 76%, but the detection rate of the 7 combined assays was 100%. For 38 culture collection strains, including clinical isolates, each individual LFTS assay had a detection rate of only 18% to 76%, but the 7 assays in combination had a detection rate of 89%. A feasibility study conducted on 20 types of meat (beef, chicken and pork) indicated that the LFTS assays detected coliform bacteria from 3 types of meat without incubation and from all other meats after 8 h of incubation. LFTS assays showed a positive signal when the meat was spoiled by more than 4.9 log10 (cfu/g) coliform bacteria. A longer incubation time led to increased bacterial counts, more positive LFTSs (1.8 at 8 h and 4.6 at 24 h) and a greater maximal signal intensity (1, 366 at 8 h and 2, 678 at 24 h). Thus, LFTSs of coliform bacteria have great potential for the rapid determination of food freshness as well as food sanitation status.

Detection in raw cow's milk of coliform bacteria - reservoir of antibiotic resistance

Antibiotic resistance is one of the main threat for people, especially since food products are more frequently identified as a reservoir of antibiotic resistance genes. Thus, the aim of the study was to analyse the Polish raw cow's milk in terms of the presence of coliform bacteria resistant to antibiotic as well as to identify volatile compounds (VC) characteristic for such bacteria. Further, the identified profile of VC could be used in development of coliform bacteria detection method directly in the product. The obtained data showed that among coliform bacteria strain isolated from Polish raw cow's milk 88% were resistant to penicillin, 39% to kanamycin, 43% to streptomycin, 78% to chloramphenicol and 55% to tetracycline. Moreover, tetM gene, frequently connected with mobile genetic elements, were detected in 42% of isolated coliform strains. Due to widespread of coliform bacteria resistant to antibiotic in raw milk, simple methods of detection of coliform bacteria in such product are required. Performed analysis suggests that lack of methional might be used as an indicator of raw cow's milk contamination with coliform bacteria.

The Likelihood of Coliform Bacteria in NJ Domestic Wells Based on Precipitation and Other Factors.

The influence of precipitation on coliform bacteria detection rates in domestic wells was investigated using data collected through the New Jersey Private Well Testing Act. Measured precipitation data from the National Weather Service (NWS) monitoring stations was compared to estimated data from the Multisensor Precipitation Estimate (MPE) in order to determine which source of data to include in the analyses. A strong concordance existed between these two precipitations datasets; therefore, MPE data was utilized as it is geographically more specific to individual wells. Statewide, 10 days of cumulative precipitation prior to testing was found to be an optimal period influencing the likelihood of coliform detections in wells. A logistic regression model was developed to predict the likelihood of coliform occurrence in wells from 10 days of cumulative precipitation data and other predictive variables including geology, season, coliform bacteria analysis method, pH, and nitrate concentration. Total coliform (TC) and fecal coliform or Escherichia coli (FC/EC) were detected more frequently when the preceding 10 days of cumulative precipitation exceeded 34.5 and 54 mm, respectively. Furthermore, the likelihood of coliform detection was highest in wells located in the bedrock region, during summer and autumn, analyzed with the enzyme substrate method, with pH between 5 and 6.99, and (for FC/EC but not TC) nitrate greater than 10 mg/L. Thus, the likelihood of coliform presence in domestic wells can be predicted from readily available environmental factors including timing and magnitude of precipitation, offering outreach opportunities and potential changes to coliform testing recommendations.

Escherichia coli strains from ostriches and their sensitivity to antimicrobial substances.

Ostriches are bred especially for their high-quality meat. There is a lack of knowledge concerning the ostrich's microflora. Escherichia coli is a commensal microorganism of the poultry intestine, ostriches included. However, some strains may become pathogenic. This study was therefore undertaken to detect coliform bacteria in ostrich faeces and to test their antibiotic profile and sensitivity to enterocins. Faeces (n=54, 18 mixture samples from 3 different age groups of 140 ostriches) were sampled to isolate coliform bacteria. The counts of coliform bacteria varied from 5.69 ± 2.4 log10 CFU/g to 5.73 ± 2.4 CFU/g. Pure colonies were identified using MALDI-TOF MS mass spectrometry and confirmed by phenotypization. Seventy-one strains were allotted to the species E. coli. Sixty-four of those 71 strains caused hemolysis. They were mostly polyresistant to antibiotics. Thirty-two poly-resistant strains of E. coli were sensitive to enterocins. These strains were most sensitive to Ent 9296 (26 strains). Moreover, Ent EM41 produced by E. faecium EM41 (isolated from ostrich faeces) inhibited the growth of 20 strains, reaching activity of 100 AU/ml. Our results indicate the possibility of enterocins being used for prevention/reduction of coliforms. Of course, in vivo studies are also being processed.

Fast detection of coliform bacteria by means of gas chromatography-differential mobility spectrometry.

In this study, we demonstrate that the combination of an enzymatic method (based on Colilert-18 medium) and gas chromatography-differential mobility spectrometry (GC-DMS) can reduce the time required for detection of coliform bacteria (including Escherichia coli) from 18 to 2.5h. The presented method includes the incubation (~2.5h) of the sample containing coliform bacteria in Colilert-18 medium. The incubation time of 2.5h is required for the activation of the β-galactosidase enzyme. Produced during the incubation biomarker o-nitrophenol (ONP) can be detected by means of GC-DMS within just 200s. The detection limit for ONP was 45ng (on-column). The method developed in this work provides significantly shorter analysis time compared with standard methods, and can be potentially adapted to the field conditions. Therefore, this method is a promising tool for an early detection of coliform bacteria (including E. coli). Graphical Abstract Fast detection of coliform bacteria by means of GC-DMS.

Prevalence of contagious and environmental mastitis-causing bacteria in bulk tank milk and its relationships with milking practices of dairy cattle herds in São Miguel Island (Azores)

This study aimed to assess the degree of contamination of bulk tank milk (BTM) by Staphylococcus spp. and coliform bacteria and to identify major milking practices that help perpetuate them in dairy cattle herds in São Miguel Island. In July 2014, BTM was sampled and a survey concerning local milking practices was conducted on 100 herds. Semi quantitative multiplex polymerase chain reaction detected coagulase-negative staphylococci, Escherichia coli, Staphylococcus aureus, and other coliform bacteria (Klebsiella oxytoca, Klebsiella pneumoniae, and Serratia marcescens) in 100, 75, 59, and 35 % of BTM, respectively. According to multivariable univariate models, on herds not using hot water for cleaning the milking machine and teat liners, there was at least 3.4 more odds (P < 0.01) to have S. aureus or coliform bacteria contamination in BTM. The likelihood of finding S. aureus in BTM was higher (P < 0.001) on herds without high hygiene during milking, when milking mastitic cows at the end, on abrupt cessation of milking at dry-off, and official milk control implementation. The glove use also favored (odds ratio (OR) 5.8; P < 0.01) the detection of coliform bacteria in BTM. Poor milking practices identified in this study should be avoided in order to decrease S. aureus and coliform bacteria contamination of BTM. Other factors associated with milk quality in São Miguel Island also should be further investigated.

Gas Chromatography–Differential Mobility Spectrometry and Gas Chromatography–Mass Spectrometry for the detection of coliform bacteria

The use of miniaturized Gas Chromatography–Differential Mobility Spectrometry (GC–DMS) is shown for the detection and identification of coliform bacteria (including Escherichia coli) grown in five different media: Colilert®-18, glucose broth, M9-medium, tryptophan broth, and tryptic soy broth. After incubation in the different media, headspace containing the volatile compounds was analyzed by the GC–DMS and the results were validated by Gas Chromatography–Mass Spectrometry (GC–MS). Results showed that the GC–DMS and GC–MS were able to detect o-nitrophenol released by coliform bacteria incubated in Colilert®-18. In addition to that, GC–MS was able to detect indole compound released by coliform bacteria grown in all media. Neither GC–DMS nor GC–MS could detect 4-methylumbelliferone from the headspace of E. coli grown in media containing 4-methylumbelliferyl-β-d-glucoronide (MUG) substrate, which was available in Colilert®-18. With the miniaturized GC–DMS being portable and can be operated using ambient pressure, this method offers a potential on-site detection of coliform bacteria.

Sensor node for remote monitoring of waterborne disease-causing bacteria.

A sensor node for sampling water and checking for the presence of harmful bacteria such as E. coli in water sources was developed in this research. A chromogenic enzyme substrate assay method was used to easily detect coliform bacteria by monitoring the color change of the sampled water mixed with a reagent. Live webcam image streaming to the web browser of the end user with a Wi-Fi connected sensor node shows the water color changes in real time. The liquid can be manipulated on the web-based user interface, and also can be observed by webcam feeds. Image streaming and web console servers run on an embedded processor with an expansion board. The UART channel of the expansion board is connected to an external Arduino board and a motor driver to control self-priming water pumps to sample the water, mix the reagent, and remove the water sample after the test is completed. The sensor node can repeat water testing until the test reagent is depleted. The authors anticipate that the use of the sensor node developed in this research can decrease the cost and required labor for testing samples in a factory environment and checking the water quality of local water sources in developing countries.

Repeat Sampling and Coliform Bacteria Detection Rates in New Jersey Domestic Wells

In compliance with the New Jersey Private Well Testing Act, 78,546 wells (93,787 samples, including samples from 13,290 wells that were analyzed more than once) were analyzed for total coliform (TC) bacteria by one or more of 39 laboratories over a 10-year period. Samples containing TC bacteria were further analyzed for the presence of either fecal coliform or E. coli (FC/EC) bacteria. The large population of wells sampled multiple times permitted a systematic study of the effect of repeat sampling on coliform bacteria detection rates. The detection rate increased with the number of times wells were sampled. In bedrock, TC bacteria were detected in 21% of the population of wells analyzed only once, 33% in the population sampled twice, and 43% in the population sampled three times. It was estimated that TC bacteria would be detected in 90% of all wells if each well was analyzed 10 times. For FC/EC bacteria, it was estimated that 21 and 68 samples, respectively, would be required to reach the 50% and 90% population detection rates. In the Coastal Plain (CP), many more samples would be required to achieve the same estimated population detection rates. The population detection rate estimates were also dependent on the type of method used, the pH of the well water, and the geologic formation in which wells were located. A single sample was not sufficient to detect coliform bacteria when present in well water.