scholarly journals Multiplex Fluorogenic Real-Time PCR for Detection and Quantification of Escherichia coli O157:H7 in Dairy Wastewater Wetlands

2002 ◽  
Vol 68 (10) ◽  
pp. 4853-4862 ◽  
Author(s):  
A. Mark Ibekwe ◽  
Pamela M. Watt ◽  
Catherine M. Grieve ◽  
Vijay K. Sharma ◽  
Steven R. Lyons
Keyword(s):  
Escherichia Coli ◽  
Detection Limit ◽  
The United States ◽  
Dairy Wastewater ◽  
Pcr Assay ◽  
E Coli ◽  
Quality Of Water ◽  
Plate Counts

ABSTRACT Surface water and groundwater are continuously used as sources of drinking water in many metropolitan areas of the United States. The quality of water from these sources may be reduced due to increases in contaminants such as Escherichia coli from urban and agricultural runoffs. In this study, a multiplex fluorogenic PCR assay was used to quantify E. coli O157:H7 in soil, manure, cow and calf feces, and dairy wastewater in an artificial wetland. Primers and probes were designed to amplify and quantify the Shiga-like toxin 1 (stx1) and 2 (stx2) genes and the intimin (eae) gene of E. coli O157:H7 in a single reaction. Primer specificity was confirmed with DNA from 33 E. coli O157:H7 and related strains with and without the three genes. A direct correlation was determined between the fluorescence threshold cycle (CT ) and the starting quantity of E. coli O157:H7 DNA. A similar correlation was observed between the CT and number of CFU per milliliter used in the PCR assay. A detection limit of 7.9 × 10−5 pg of E. coli O157:H7 DNA ml−1 equivalent to approximately 6.4 × 103 CFU of E. coli O157:H7 ml−1 based on plate counts was determined. Quantification of E. coli O157:H7 in soil, manure, feces, and wastewater was possible when cell numbers were ≥3.5 × 104 CFU g−1. E. coli O157:H7 levels detected in wetland samples decreased by about 2 logs between wetland influents and effluents. The detection limit of the assay in soil was improved to less than 10 CFU g−1 with a 16-h enrichment. These results indicate that the developed PCR assay is suitable for quantitative determination of E. coli O157:H7 in environmental samples and represents a considerable advancement in pathogen quantification in different ecosystems.

scholarly journals The quantitative bacteriology of some commercial bivalve shellfish entering British markets

1975 ◽  
Vol 74 (3) ◽  
pp. 431-440 ◽  
Author(s):  
P. A. Ayres
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Health Risk ◽  
E Coli ◽  
Accepted Standard ◽  
Plate Counts ◽  
Bivalve Shellfish ◽  
Faecal Streptococci ◽  
Specific Illness

SUMMARYIncidents of non-specific illness associated with the consumption of oysters have highlighted the lack of published information on the bacteriology of shellfish suitable for consumption. Investigations showed that the majority of molluscan shellfish entering English markets conform to the accepted standard of less than 5 Escherichia coli/ml. tissue. The numbers of E. coli were related to the sanitary quality of the growing area but no relation could be established between numbers of E. coli and coliforms, faecal streptococci or Clostridium welchii. The numbers of non-specific bacteria varied considerably but shellfish from sources associated with non-specific illness yielded relatively high counts at 37° C. The results showed that there was no justification for a standard based on total plate counts, which often exceeded 106/g. Such a standard would have to be coupled with spoilage or the incidence of non-specific illness. The relation between the numbers of non-specific bacteria growing at 20 and 37° C. appears to be a useful measure for assessing the likelihood that raw shellfish are a public health risk.

A Bacteriological Survey of Raw Ground Beef1

1977 ◽  
Vol 40 (11) ◽  
pp. 790-794 ◽  
Author(s):  
JAMES F. FOSTER ◽  
JAMES L. FOWLER ◽  
WARREN C. LADIGES
Keyword(s):  
Escherichia Coli ◽  
Microbiological Quality ◽  
Retail Store ◽  
Most Probable Number ◽  
Probable Number ◽  
E Coli ◽  
Fecal Streptococci ◽  
Plate Counts ◽  
Aerobic And Anaerobic

The microbiological quality of 150 units of raw ground beef obtained from a local retail store was determined. The range of aerobic plate counts was from 6.9 × 104 to 8.3 × 107/g. By using the most probable number method 96.7% of the 150 units were positive for coliforms, 94.7% for Escherichia coli and 61.3% for Staphylococcus aureus. By the plate methods, 99.3% of the units were positive for fecal streptococci and 56% were positive for Clostridium perfringens. No salmonellae were isolated. Aerobic and anaerobic organisms were isolated and identified. E. coli was the most frequently isolated aerobe followed by organisms in the Klebsiella-Enterobacter group. Among the anaerobic isolates, C. perfringens was the organism most frequently encountered.

Microbiological Quality of Frozen Cream-Type Pies Sold in Canada

1983 ◽  
Vol 46 (1) ◽  
pp. 34-40 ◽  
Author(s):  
E. C. D. TODD ◽  
G. A. JARVIS ◽  
K. F. WEISS ◽  
G. W. RIEDEL ◽  
S. CHARBONNEAU
Keyword(s):  
United States ◽  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Microbiological Quality ◽  
The United States ◽  
National Guidelines ◽  
E Coli ◽  
Yeasts And Molds

Ten types of frozen cream-type pies, manufactured in Canada and imported from the United States, were analyzed for aerobic colony counts, yeasts and molds, coliforms, Escherichia coli, Staphylococcus aureus and Salmonella. The variations in counts depended more on the manufacturer than on the type of pie and the ingredients used. Five of the 465 examined pies had an excess of 105 aerobic colony counts/g, whereas the median value for all the pies examined was between 102 and 103 CFU/g. E. coli and S. aureus were present in few pies, mainly made by one manufacturer, but there was no correlation between high aerobic colony counts and these organisms. Salmonella was not found in any of the pies. Percentage distributions of the estimated ‘population’ of pies available nationally at the time of the survey were statistically determined. These were then compared with suggested national guidelines in the form of a three-class acceptance plan based on United States surveys and desirable manufacturing practices. These indicate that pies should contain aerobic colony counts of <50,000/g, yeast and mold counts of <500/g, S. aureus counts of <100/g, coliform counts of <50/g, E. coli counts of <10/g, and no Salmonella. Three of the six manufacturers would have had an estimated 5.4 to 32.6% of lots in excess of the guidelines at the time of the survey.

Bacteriological Survey of Frozen Meat Ravioli Produced at Establishments Under Federal Inspection

1983 ◽  
Vol 46 (8) ◽  
pp. 710-713
Author(s):  
DOUGLAS F. CAMPBELL ◽  
MARTHA Y. WORKMAN ◽  
GEORGE W. KRUMM ◽  
RALPH W. JOHNSTON
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Production Line ◽  
Microbiological Quality ◽  
Slow Freezing ◽  
Geometric Means ◽  
E Coli ◽  
Plate Counts ◽  
Product Sets

During visits to 20 federally inspected establishments producing meat ravioli, 577 production line samples and 480 finished product units were collected for bacteriological analyses. Four types of finished, packaged ravioli were encountered: (a) whole ravioli boiled at least 5 min; (b) raw pasta stuffed with a cooked filling; (c) only the meat component cooked; and (d) uncooked ravioli. The microbiological quality of frozen ravioli was affected more by the filling than the pasta. Slow freezing resulted in increased bacterial levels in the finished product. For the boiled ravioli, 100% of the finished product sets contained less than 50 coliforms per g, four of five sets had less than one Escherichia coli per g, and 100% had fewer than one Staphylococcus aureus per g. Four of five sets of packaged boiled ravioli had aerobic plate counts (APC) of less than 10,000 per g. For the raw pasta with a cooked filling-type ravioli, the geometric means of 9 sets were: coliforms, 47 per g; E. coli, 6.7 per g; S. aureus, 10 per g; and APC, 170,000 per g. For the ravioli with only the meat component cooked, the geometric means of 27 sets were: coliforms, 190 per g; E. coli, 1.8 per g; S. aureus, 3.9 per g; and APC, 300,000 per g. For uncooked ravioli, the geometric means of 7 sets were: coliforms, 490 per g; E. coli, 19 per g; S. aureus, 5.7 per g; and APC, 690,000 per g. Only one finished ravioli unit in 480 contained Salmonella.

COLIFORM BACTERIA IN SEA WATER AND SHELLFISH: II. THE E. C. CONFIRMATION TEST FOR ESCHERICHIA COLI

1961 ◽  
Vol 7 (5) ◽  
pp. 733-739 ◽  
Author(s):  
A. D. Tennant ◽  
J. E. Reid ◽  
L. J. Rockwell ◽  
E. T. Bynoe
Keyword(s):  
Escherichia Coli ◽  
Sea Water ◽  
The United States ◽  
Coliform Bacteria ◽  
Simple Test ◽  
E Coli ◽  
Confirmation Test ◽  
The Mean ◽  
Test Cultures

Recently there has been considerable interest in Canada and the United States in the development of an effective, simple test for the estimation of Escherichia coli densities in shellfish and shellfish-growing waters as a better indication of "faecal" pollution than that provided by the coliform group. The fidelity of the E.C. confirmation test for the determination of E. coli densities was evaluated in 15 shellfish-growing areas which were classified as "polluted" or "unpolluted" according to generally accepted criteria. Coliform strains were isolated from 2765 E.C. gas positive confirmation test cultures and identified by IMViC tests. The mean fidelity of the test, as an indication of the presence of E. coli, was circa 90 per cent; contrary to expectations, however, the per cent recovery of E. coli was higher in unpolluted areas than in polluted areas, and varied considerably from region to region. Aerobacter aerogenes types I and II were the most common other E.C. gas positive coliform biotypes found. The usefulness of the confirmation test and the sanitary significance of these data are discussed.

scholarly journals Salmonella and Escherichia coli O157:H7 Contamination on Hides and Carcasses of Cull Cattle Presented for Slaughter in the United States: an Evaluation of Prevalence and Bacterial Loads by Immunomagnetic Separation and Direct Plating Methods

2008 ◽  
Vol 74 (20) ◽  
pp. 6289-6297 ◽  
Author(s):  
Dayna M. Brichta-Harhay ◽  
Michael N. Guerini ◽  
Terrance M. Arthur ◽  
Joseph M. Bosilevac ◽  
Norasak Kalchayanand ◽  
...  
Keyword(s):  
United States ◽  
Escherichia Coli ◽  
Geometric Mean ◽  
Immunomagnetic Separation ◽  
The United States ◽  
E Coli ◽  
Processing Plants ◽  
Pathogen Prevalence ◽  
Plate Counts ◽  
Carcass Contamination

ABSTRACT The hide and carcass hygiene of cull cattle at slaughter in four geographically distant regions of the United States was examined from July 2005 to April 2006 by measuring the aerobic plate counts (APC) and the prevalences and loads of Salmonella and Escherichia coli O157:H7. The geometric mean log10 APC CFU/100 cm2 levels on hides and preevisceration and postintervention carcasses ranged from 6.17 to 8.19, 4.24 to 6.47, and 1.46 to 1.96, respectively, and were highest in the summer (P < 0.0001). The average prevalences of Salmonella on hides and preevisceration and postintervention carcasses were 89.6% (95% confidence interval [CI], 85.1 to 94.0), 50.2% (95% CI, 40.9 to 59.5), and 0.8% (95% CI, 0.18 to 1.42), respectively. The prevalences of E. coli O157:H7 were 46.9% (95% CI, 37.3 to 56.6) and 16.7% (95% CI, 9.8 to 23.6) on hides and preevisceration carcasses, respectively. Examination of the concomitant incidence of Salmonella and E. coli O157:H7 showed that, on average, 33.3% (95% CI, 15.9 to 69.8) of cattle hide and 4.1% (95% CI, 0.98 to 17.3) of preevisceration carcass samples were contaminated with both pathogens. The pathogen prevalence on hides and carcasses was not significantly affected by the season; however, significant differences were observed between plants with respect to the incoming pathogen load and the ability to mitigate hide-to-carcass transfer. In spite of these differences, postintervention carcass contamination was significantly reduced (P < 0.001), likely as a result of the use of one or more of the processing interventions employed at each of the four processing plants examined.

scholarly journals Physicochemical quality of and Escherichia coli resistance profiles in urban surface waters

2020 ◽  
Vol 80 (3) ◽  
pp. 661-668
Author(s):  
I. Malagi ◽  
S. C. Sampaio ◽  
F. G. S. Pinto ◽  
D. M. Rosa ◽  
R. R. dos Reis
Keyword(s):  
Escherichia Coli ◽  
Public Health Problem ◽  
Resistant Bacteria ◽  
Urban Centers ◽  
E Coli ◽  
Quality Of Water ◽  
Microbiological Parameters ◽  
National Environment ◽  
Thermotolerant Coliforms

Abstract Aquatic ecosystems of urban rivers are contaminated through waste disposal, which poses a public health problem. The objective of this research was to evaluate the quality of water used for recreation and public supply of six rivers in the city of Cascavel - Paraná, including Cascavel, Quati, Bezerra, Antas, Clarito and Amambay. Samples were collected every 4 months in 2017, and their physicochemical and microbiological parameters, as well as resistance profiles of strains of Escherichia coli to antimicrobials distributed by pharmacies of the primary healthcare network, were evaluated. Parameters such as water temperature, turbidity, total nitrogen, total phosphorus, total coliforms and thermotolerant coliforms showed significant differences. The allowed limit for thermotolerant coliforms, which was set by National Environment Council, Resolution 357/2005, was exceeded in all of the six analyzed rivers. It was determined that 48.1% of E. coli strains showed resistance to nine antimicrobial tested. The highest levels of resistance were found for ampicillin (27.7%), tetracycline (27.7%) and amoxicillin (24.0%). The results of this study contribute to the understanding of the hazards associated with the contamination of springs in urban centers with wastewater containing resistant bacteria. Therefore, recovery work is necessary in these areas because of the importance of these water sources for the entire western region of Paraná state.

Assessment of Microbiological Safety and Quality of Marinades Used To Treat Beef and That Were Collected over a 12-Month Period from Specialty Retailers Near Raleigh, North Carolina

2018 ◽  
Vol 81 (3) ◽  
pp. 490-496 ◽  
Author(s):  
Yangjin Jung ◽  
Christopher L. Rupert ◽  
Benjamin Chapman ◽  
Anna C. S. Porto Fett ◽  
John B. Luchansky
Keyword(s):  
Escherichia Coli ◽  
North Carolina ◽  
Real Time ◽  
Shiga Toxin ◽  
Real Time Pcr ◽  
Plate Count ◽  
Pcr Assay ◽  
E Coli ◽  
Aerobic Plate Count

ABSTRACT In total, 115 marinade samples (58 fresh marinades and 57 spent marinades) were collected over 12 months from specialty retailers (four individual stores) near Raleigh, NC. These marinades were screened for total mesophilic aerobic plate count (M-APC), total psychrotrophic aerobic plate count (P-APC), and Enterobacteriaceae. These marinades were also screened for the seven regulated serogroups of Shiga toxin–producing Escherichia coli. Stores A and B used immersion to marinade raw beef cuts, whereas stores C-1 and C-2 used vacuum tumbling. In general, marinade temperatures at the stores ranged from 1.8 to 6.6°C, and beef cuts were marinated from a few minutes to up to 3 days. Regardless of the process used to marinade meat, levels of M-APC and P-APC in fresh marinades ranged from 3.4 to 4.7 and 1.4 to 1.8 log CFU/mL, respectively, whereas Enterobacteriaceae were not detected in any fresh marinades, even after enrichment. However, levels of M-APC, P-APC, and Enterobacteriaceae in spent marinades collected from stores C-1 and C-2 (ca. 3.6 to 7.1 log CFU/mL) were significantly higher (P &lt; 0.05) compared with levels of these same types of bacteria enumerated from spent marinades collected at stores A and B (ca. ≤0.7 to 4.9 log CFU/mL). None of the 115 marinade samples tested positive for Shiga toxin–producing E. coli by using a BAX system real-time PCR assay. No significant (P &gt; 0.05) association was observed between microbial levels (i.e., M-APC, P-APC, and Enterobacteriaceae) and the temperature or duration of the marination process. Levels of M-APC, P-APC, and Enterobacteriaceae in spent marinades were significantly affected by the marination method (P &lt; 0.05), with levels, in general, being higher in marinades used for tumbling. Thus, retailers must continue to keep marinade solutions and meat at a safe temperature (i.e., ≤4°C) and to properly and frequently sanitize the equipment and environment in both the processing area and deli case.

scholarly journals Quality of water treated with Moringa oleifera Lam seeds

2020 ◽  
Vol 9 (4) ◽  
pp. e164943041
Author(s):  
Semirames do Nascimento Silva ◽  
Eliezer da Cunha Siqueira ◽  
Josivanda Palmeira Gomes ◽  
Roberta de Oliveira Sousa Wanderley ◽  
Polyana Barbosa Silva
Keyword(s):  
Escherichia Coli ◽  
Moringa Oleifera ◽  
E Coli ◽  
Quality Of Water ◽  
Moringa Oleifera Lam

O objetivo deste trabalho foi avaliar a qualidade química e microbiológica da água de cisternas antes e após o tratamento com sementes de Moringa ou leifera Lam. Como sementes de moringa usadas no trabalho foram compostas no município de Cajazeiras. O coagulante da moringa foi usado em meio aquoso. A água filtrada na obra veio de cisternas construídas no assentamento de Santo Antônio, município de Cajazeiras. A água utilizada foi analisada antes e após o tratamento.Uma análise dos parâmetros: pH, condutividade elétrica, alcalinidade total, provável resíduo e total de dissolvidos foi realizada no Laboratório de Água e Solo e no Laboratório de Microbiologia do Instituto Federal de Educação, Ciência e Tecnologia do Campus da Paraíba, análises de coliformes fecais e Escherichia coli foram mantidos. Todas as estatísticas de substâncias químicas mostram diferenças antes e depois do tratamento. Os controles C1 e C5 apresentam presença de Escherichia coli , os demais são causados pela E. coli .

scholarly journals Microbiological quality of seasoned chicken cuts using Escherichia coli and Salmonella spp. as quality indicators.

2020 ◽  
Vol 9 (11) ◽  
pp. e38091110013
Author(s):  
Flávio Barbosa da Silva ◽  
Bruna Ribeiro Arrais ◽  
Marcos Roberto Alves Ferreira ◽  
Iderval da Silva Júnior Sobrinho ◽  
Márcia Dias ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Microbiological Quality ◽  
Poultry Production ◽  
Salmonella Spp ◽  
Production Chain ◽  
E Coli ◽  
Important Health ◽  
Pathogenic Serotypes

Enteropathogens such as Salmonella spp. and Escherichia coli are important health challenges in the poultry production chain, because when installed in the production chain, they impair the safety of food supply. The determination of the microbiological quality of chicken meat, especially when marketed in spiced form, is necessary, given that consumer prefer this type of processed protein. This work aims to evaluate the microbiological quality of the meat of seasoned chicken marketed in the municipality of Rio Verde, Goias, using Salmonella spp. and Escherichia coli as target microorganisms, and considering as variables the type of establishment, validity date and presence of municipal inspection seal. From 80 analyzed samples, 30% (24/80) of samples were positive for Salmonella spp. and 55% (44/80) for E. coli. Regarding the type of establishment, it was observed that 27.45% (14/51) of supermarket samples and 34.48% (10/29) of meat store samples were contaminated by Salmonella spp. Considering E. coli, 49.01% (25/51) of supermarket samples and 65.55% (19/29) of meat store samples were positive for this pathogen. 80% (64/80) of the samples had the municipal inspection seal (MIS), and 83.75% (67/80) contained the expiration date on the label. The legislation in its narrative guarantee’s protection for the consumer with regard to the presence of pathogenic serotypes of these agents, however, it is worth noting that the effective action of health surveillance, and the constant laboratory investigation of the products are necessary.

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