Inactivation of Escherichia coli O157:H7 and Salmonella enteritidis in Liquid Egg White Using Pulsed Electric Field

ABSTRACT: The effects of temperature and pulsed electric field (PEF) intensity on inactivation of pathogens such as Escherichia coli O157:H7 and Salmonella enteritidis in egg white was investigated. Liquid egg white inoculated with 10⁸ colony-forming units (CFU)/mL of each pathogen was treated with up to 60 pulses (each of 2 JAS width) at electric field intensities of 20 and 30 kV/cm. The processing temperatures were 10°C, 20°C, and 30°C. After treatment, uninjured and total viable cells were enumerated in selective and nonselective agars, respectively. Maximum inactivations of 3.7 and 2.9 log units were obtained for S. enteritidis and E. coli O157:H7, respectively, while injured cells accounted for 0.5 and 0.9 logs for E. coli O157:H7 and S. enteritidis , respectively. For both bacteria, increasing treatment temperature tended to increase the inactivation rate. There was synergy between electric field intensity and processing temperature. The inactivation rate constant k _T values for E. coli O157:H7 on both selective and nonselective agars were 8.2 × 10^-3 and 6.6 × 10^-3/μS, whereas the values for S. enteritidis were 16.2 × 10^-3 and 12.6 × 10^-3/μS, respectively. The results suggest that E. coli O157:H7 was more resistant to heat-PEF treatment compared with S. enteritidis.     

EXAMINATION OF THE MICROBIAL STATUS OF SELECTED INDIGENOUS FERMENTED FOODS IN NIGERIA

Four Nigerian traditionally fermented foods (wara, nono, ogi and kununzaki) were evaluated for the presence of some microorganisms of public health concern. Among the dairy foods , Staphylococcus aureus and Klebsiella sp. were isolated from wara while Escherichia coli, Salmonella sp. and Klebsiella sp. were isolated from nono. The cereal-based fermented foods (ogi and kunu-zaki) contained Bacillus subtilis, E. coli, S. aureus, Klebsiella sp. and Enterococcus faecalis. The mesophilic aerobic counts were: 5 × 10⁵ for wara; nono, 1.53 × 10⁷; ogi, 3.6 × 10⁶ and kunu-zaki, 2.6 × 10⁶ cfu/mL. The enterobacteriaceae counts on nono, wara, ogi and kunu-zaki were 1.79 × 10⁷, 4.5 × 10⁵, 4.0 × 10⁵ and 1.2 × 10⁶ cfu/mL, respectively. No Vibrio count (detection limit: <10 cfu/mL) was recorded in all the food samples considered. The yeast and mold counts ranged from 1.0 × 10⁵– 3.31 × 10⁷ among the food products. The antimicrobial susceptibility patterns of the organisms isolated from dairy products (nono and wara) revealed that they were resistant to ampicillin (100%) and sensitive to gentamicin (100%) and nalidixic acid (100%). Most isolates from cereal based products (ogi and kunu-zaki) were 100% resistant to penicillin, ampicillin and chloramphenicol. This work highlights the need to maintain hygienic standards in the preparation of our locally fermented cereal and dairy foods.     

CONCURRENT DETECTION OF ESCHERICHIA COLI O157:H7 AND SALMONELLA FROM A SINGLE ENRICHMENT IN 24 H

The objective of this study was to determine if a single assay protocol could result in the concurrent detection of Escherichia coli O157:H7 and Salmonella from a single sample grown in a single enrichment in 24 h. Twenty-five and 375 g of ground beef nonfat dry milk, and dry pet food samples were seeded with low (10 cfu/sample) and high (100 cfu/sample) levels ofE. coli O157:H7 and Salmonella cultures and incubated at 35 and 41C for 18 h for nonselective preenrichment. Incubated samples were analyzed by immunomagnetic separation (IMS) following a 6 h incubation for selective enrichment at 37C using M-broth and enzyme linked immumosorbent assay (ELISA). Depending on the food samples and the inoculation level, the minimum concurrent detection level of E. coli O157:H7 and Salmonella was <1 cfu/g in the samples at the competitor flora level of 10⁵ cfu/g or less in ground beef samples, but in other cases of higher competitor loads and low target inoculations E. coli O157:H7 could not be detected in the presence of the Salmonella.     

Decontamination Efficacy of Combined Chlorine Dioxide with Ultrasonication on Apples and Lettuce

ABSTRACT:  The bacterial reduction of Salmonella and Escherichia coli O157:H7-inoculated apples and lettuce by ClO₂ at 0, 5, 10, 20, and 40 ppm with and without 170-kHz ultrasonic treatments for 3, 6, and 10 min, respectively, have been studied. The treatments of ClO₂ at 20 and 40 ppm for 3, 6, and 10 min or at 5 and 10 ppm for 6 and 10 min with 170-kHz ultrasonication caused 3.115 to 4.253 log reductions in Salmonella and 2.235 to 3.865 log reduction in E. coli O157:H7 on inoculated apples. Using combined ClO₂ and ultrasonication to treat 4.48 × 10⁴ CFU/g Salmonella and 1.07 × 10⁵ CFU/g E. coli O157:H7-inoculated lettuce, the bacterial reductions were 2.257 to 2.972 and 1.357 to 2.264 log, respectively. The residual ClO₂ decreased with increasing treatment times, over 80% of ClO₂ was detected after the 3-min treatment, and more than 70% remained after the 10-min treatment time. No bacteria were recovered from the posttreatment solutions of ClO₂ or ClO₂ combined with ultrasonication. The temperature of the ClO₂ treatment was 20.1 °C, and it increased to 40.1, 44.9, and 50.3 °C, with 170-kHz ultrasonic treatments for 3, 6, and 10 min, respectively, on apples.     

Empirical Distribution Models for Escherichia coli 57:H7 in Ground Beef Produced by a Mid-size Commercial Grinder

ABSTRACT: The purpose of this research was to develop empirical models that describe the amount and distribution of ground beef contaminated with Escherichia coli O157:H7 when a contaminated beef trim is introduced into a batch of uncontaminated beef before processing in a mid-size commercial grinder (34 g/s). A beef trim was inoculated with a rifampacin-resistant strain of E. coli O157:H7 and added to a batch of noncontaminated trims at the grinding step. To study the distribution of the E. coli O157:H7^rif in the ground beef, 6 treatments with different inoculum levels (1 to 6 log₁₀ colony-forming units [CFU]) were tested. Removal or pick up of the residual contamination with E. coli O157:H7^rif left in the grinder was evaluated. E. coli O157:H7^rif was detected in 9% to 86% of the total ground beef for the 1 to 6 log₁₀ CFU inoculum levels, respectively. E. coli O157:H7^rif contamination was detected in the collar that fixes the grinder's die and blade to the hub. An exponential algorithm described the relationship between the quantities of ground beef containing E. coli O157:H7^rif and the inoculum level ( R ²= 0.82). Distribution models based on a Chi-squared algorithm were developed for each inoculum level describing the contamination level as a function of the batch fraction processed ( R ²= 0.81 to 0.99). The results of this study corroborate that when beef processors test for pathogenic contamination in a mid-scale grinder, they should test the beef residues in the collar that fixes the grinder's die and blade to the hub.     

Piezoelectric Flow Injection Analysis Biosensor for the Detection of Salmonella Typhimurium

ABSTRACT: Piezoelectric biosensors have the potential to provide direct detection of food contaminants, such as pathogens. In this study, Protein A antibody immobilization was used for the activation of the piezoelectric biosensor to detect Salmonella typhimurium. The overall system consisted of a new design for a flow cell and flow injection analysis system. The flow cell made possible a baseline stability of ± 1 Hz out of 5 MHz for hours. The sensor had responses of 5 to 65 Hz in 30 min with R²= 0.95 for S. typhimurium concentrations of 10⁷to 10⁹ CFU/ml under continuous flow, and 3 to 75 Hz in 40 min with R²= 0.96 for S. typhimurium concentrations of 10⁶ to 10¹⁰ CFU/ ml under stop flow. Cross-reactivity tests were performed with nonpathogenic Escherichia coli, Escherichia coli O157:H7, Listeria monocytogenes , and Vibrio parahaemolyticus and showed less than 10% response.     

Viability of probiotic micro-organisms (Lactobacillus casei Shirota and Bifidobacterium animalis subspp. lactis) in a milk-based dessert with cranberry sauce

The objective of this study was to evaluate the survival of two probiotic micro-organisms, Lactobacillus casei Shirota and Bifidobacterium animalis subspp . lactis , in a milk-based dessert (2.7% fat) with cranberry sauce added. B. lactis had a final concentration of 1.99 × 10⁶ cfu/g after 21 days of study, with a logarithmic decrease of 8.87%. On other hand, L. casei Shirota had a final concentration of 2.05 × 10⁷ cfu/g at the end of the same period, a logarithmic decrease of 8.41%. Statistical analysis showed that significant differences existed between both micro-organisms and over various storage times, the more viable micro-organism being L. casei Shirota , which decreased less than one logarithmic cycle after 21 days.     

Immunoliposomes Sandwich Fluorometric Assay (ILSF) for Detection of Escherichia coli O157:H7

ABSTRACT: We report the development of automated flourometric immunoassay for the detection of Escherichia coli O157:H7, using antibody-directed liposomes (immunoliposomes) encapsulating fluorophore as an analytical reagent. Thiolated antibodies (anti- E. coli O157:H7) were coupled to malemide-tagged liposomes encapsulating dye. To automate the assay, a fluorescence plate reader was included in the assay system to detect fluorophore released from lysed liposomes in a microplate. The detection limit of the current assay with pure cultures of the serotype was about 10⁴ colony-forming units (CFU)/mL. The assay can detect E. coli O157 in ground beef samples inoculated with as few as 0.8 CFU/mL after a 12-h enrichment. These results demonstrate the feasibility of using fluorophore-encapsulated immunoliposomes in a microtiter plate for the rapid and automated detection of molecules with multivalent antigenic sites.     

APPLICATION OF CHLORINE TO REDUCE POPULATIONS OF ESCHERICHIA COLI ON BEEF

The effects of chlorine against 2 strains of E. coli attached to the surface of beef carcass tissue (BCT) were examined using a model carcass washer. Lean and adipose BCT with approximately 5 log ₁₀ CFU/cm ² E. coli bacteria were spray-treated with water and sodium hypochlorite (NaOCl) to give chlorine concentrations of 50, 100, 250, 500, or 800ppm, incubated for 24 h, 4C, and E. coli populations enumerated. Spray treatments with water did significantly (P < 0.05) reduce the bacterial populations of either organism attached to lean or adipose BCT, as compared to populations of controls; however, reductions were less than 0.60 log ₁₀ CFU/cm ². Treatments with 500 and 800 ppm chlorine against E. coli ATCC 25922 attached to BCT resulted in the greatest reductions of 1.22 and 1.28 log ₁₀ CFU/cm ², respectively. At 800 ppm chlorine , E. coli O157:H7 ATCC 43895 attached to BCT was reduced by 1.04 log ₁₀ CFU/cm ², whereas spray treatments with 50, 100, 250, and 500 ppm chlorine resulted in reductions of < 1 log ₁₀ CFU/cm ². Spray treatments with chlorine from sodium hypochlorite solutions reduced populations of E. coli, however, these reductions were not sufficient to completely inactivate the bacteria attached to red meat .     

The distribution of Enterobacteriaceae and some pathogenic micro-organisms in nonbranded white cheese samples sold in Ankara city

In this research, 75 samples of nonbranded white cheese, presented for sale in small markets from various regions and bazaars in Ankara, were studied. Eighty-six isolates, 71 of which are Gram-negative isolates and 15 of which are Gram-positive isolates, were obtained. These isolates were identified by using bioMérieux API20E and classical methods. Total mesophilic bacteria counts and total coliform bacteria counts were carried out for each white cheese sample. The total average mesophilic bacteria of 75 white cheese samples was 15.5 × 10⁵ cfu/g and the total average coliform bacteria were 13.6 × 10⁵ cfu/g.     

MICROBIOLOGICAL QUALITY OF TRADITIONAL MARKET CURED FISH IN TANZANIA

The microbiological quality of six varieties of retail market traditionally cured fish in Morogoro, Tanzania was investigated over a five-month period. The fish were contaminated with bacteria and molds at levels of: total aerobes, 10⁶ - 1.7 × 10⁷ c.f.u/g; faecal coliforms, 1.1 × 10¹ - 2.5 × 10³ MPN/g; faecal streptococci, 1.4 × 10¹ - 1.3 × 10³ MPN/g; Staphylococcus aureus, 1.3 × 10³ - 8.6 × 10³ c.f.u/g; Aspergillus flavus group, 2.1 × 10¹ - 2 × 10² c.f.u/g of fish. Of faecal coliform, 45% of the isolates were Escherichia coli. Twenty-five percent of the S. aureus isolates were coagulase positive. Sixteen percent of A. flavus isolates were aflatoxigenic. Aflatoxin contamination ranged from O to 18.5 μg/kg of fish. Insect infestation by Dermestes spp. and mites was observed. The results of this preliminary study emphasize the importance of proper processing and handling offish in the tropics in order to safeguard public health .     

RADIATION STERILIZATION OF SPICES FOR HOSPITAL FOOD SERVICES AND PATIENT CARE

A survey of the commercial spices used by food services in a typical hospital environment revealed high contamination with microorganisms, i.e., 10⁴ to 10⁷ counts per gram. The predominant microorganisms were as followed (in colony counts/gram): (1) heat-resistant bacterial spores in black pepper, 1 × 10⁷; thyme, 2 × 10⁶; anise, 7 × 10⁴; curry powder, 4 × 10⁵; poultry seasoning, 8 × 10⁴; pickling spice, cardamom, and cumin, 1.5–3 × 10⁴; (2) mixed populations of vegetative cells and bacterial spores in cumin, 1 × 10⁶; (3) molds in cream of tartar, 2 × 10⁴. Sterility of food may be important in a hospital setting, especially in the care of immunocompromised patients. To eliminate the organisms, we recommend radiation treatment, accompanied by appropriate microbiological quality control. On the basis of radiation survival data, the composite natural flora would be reduced to the level of "commercial sterility" (defined as less than 10 organisms per gram((Kiss 1982) by the following minimum radiation doses (in kGy): black pepper, 13; thyme, 13; cumin, 12; anise, 10; curry, 7.3; pickling spice, 7; poultry seasoning, 6; cardamom, 9.4; cream of tartar, 4. For practical purposes, two dose levels can be recommended for treatment of spices in the hospital environment, low = 6–10 kGy and high = 10–15 kGy.     

Antimicrobial effect of electrolyzed oxidizing water against Escherichia coli O157:H7 and Listeria monocytogenes on fresh strawberries (Fragaria x ananassa)

ABSTRACT:  Antibacterial activity of electrolyzed oxidizing (EO) water prepared from 0.05% or 0.10% (w/v) sodium chloride (NaCl) solutions against indigenous bacteria associated with fresh strawberries ( Fragaria × ananassa ) was evaluated. The efficacy of EO water and sodium hypochlorite (NaOCl) solution in eliminating and controlling the growth of Listeria monocytogenes and Escherichia coli O157:H7 inoculated onto strawberries stored at 4 ± 1 °C up to 15 d was investigated at exposure time of 1, 5, or 10 min. Posttreatment neutralization of fruit surfaces was also determined. More than 2 log₁₀ CFU/g reductions of aerobic mesophiles were obtained in fruits washed for 10 or 15 min in EO water prepared from 0.10% (w/v) NaCl solution. Bactericidal activity of the disinfectants against L. monocytogenes and E. coli O157:H7 was not affected by posttreatment neutralization, and increasing exposure time did not significantly increase the antibacterial efficacy against both pathogens. While washing fruit surfaces with distilled water resulted in 1.90 and 1.27 log₁₀ CFU/mL of rinse fluid reduction of L. monocytogenes and E. coli O157:H7, respectively, ≥ 2.60 log₁₀ CFU/mL of rinse fluid reduction of L. monocytogenes and up to 2.35 and 3.12 log₁₀ CFU/mL of rinse fluid reduction of E. coli O157:H7 were observed on fruit surfaces washed with EO water and NaOCl solution, respectively. Listeria monocytogenes and E. coli O157:H7 populations decreased over storage regardless of prior treatment. However, EO water and aqueous NaOCl did not show higher antimicrobial potential than water treatment during refrigeration storage.     

Combination of Carbon Dioxide and Cinnamon to Inactivate Escherichia coli O157:H7 in Apple Juice

ABSTRACT: Pasteurized apple juice with CO2 (0, 1, and 4%) and cinnamon (0 and 0.3%) was inoculated with Escherichia coli O157:H7 at 104 CFU/mL, and stored at 5 and 20 °C. Counts on nonselective and selective media, and thin agar layer (TAL; selective medium overlaid with nonselective medium) were determined at 1 h and 1, 3, 7, and 14 d. Inactivation was greater at 20 °C. Samples with 1 and 4% CO₂, alone and combined with cinnamon, presented < 0.7 log CFU/mL in 3 d. Counts in apple juice inoculated at 10² CFU/mL, a low-level E. coli O157:H7 contamination, were nondetectable at 3 d. The TAL method was as effective as nonselective medium to recover injured cells.     

Effect of technological parameters on the characteristics of kasseri cheese made from raw or pasteurized ewes' milk

Two groups of kasseri cheese (pasta filata type) were manufactured from raw or pasteurized ewes' milk, without starter cultures. Cheeses of each group were divided into two subgroups: the first was ripened and stored at 4°C and packaged in plastic film; the second ripened and stored at 15°C and coated with paraffin wax. Milk pasteurization and technological parameters had a significant effect on the pH ( P  < 0.05), while only technological parameters had an effect on the total solids content. At day 120, the range of mean cfu/g counts for the mesophilic aerobic flora was 9.5 × 10⁷−1.4 × 10⁸; for the thermophilic streptococci, the range was 2.6 × 10⁷−7.6 × 10⁷; and for the thermophilic bacilli, 9.8 × 10⁶−1.7 × 10⁷. Changes in the N fractions became significant after 30 days of ripening. For mature 120-day-old cheeses, the percentage of total N soluble at pH 4.6 was 22.7%–22.9% in raw milk cheeses and 19.0%–21.7% in pasteurized milk cheeses. The percentage of total N soluble at 12% TCA was 10.1%–12.2% in raw milk cheeses and 7.3%–11.5% in pasteurized milk cheeses; the percentages of total N soluble at 5% PTA were 3.1%–4.0% and 2.6%–3.6%, respectively. The residual α_s-casein percentages at day 120 ranged between 63% and 78% of the respective area at day 1; the residual β-casein ranged between 67% and 75%. There were some characteristic differences in the reverse phase-HPLC peptide profiles of the four cheeses. In general, the effect of the different ripening conditions was more pronounced in cheeses made from pasteurized milk.     

Edible Apple Film Wraps Containing Plant Antimicrobials Inactivate Foodborne Pathogens on Meat and Poultry Products

ABSTRACT:  Apple-based edible films containing plant antimicrobials were evaluated for their activity against pathogenic bacteria on meat and poultry products.  Salmonella enterica  or  E. coli  O157:H7 (10⁷ CFU/g) cultures were surface inoculated on chicken breasts and  Listeria monocytogenes  (10⁶ CFU/g) on ham. The inoculated products were then wrapped with edible films containing 3 concentrations (0.5%, 1.5%, and 3%) of cinnamaldehyde or carvacrol. Following incubation at either 23 or 4 °C for 72 h, samples were stomached in buffered peptone water, diluted, and plated for enumeration of survivors. The antimicrobial films exhibited concentration-dependent activities against the pathogens tested. At 23 °C on chicken breasts, films with 3% antimicrobials showed the highest reductions (4.3 to 6.8 log CFU/g) of both  S. enterica  and  E. coli  O157:H7. Films with 1.5% and 0.5% antimicrobials showed 2.4 to 4.3 and 1.6 to 2.8 log reductions, respectively. At 4 °C, carvacrol exhibited greater activity than did cinnamaldehyde. Films with 3%, 1.5%, and 0.5% carvacrol reduced the bacterial populations by about 3, 1.6 to 3, and 0.8 to 1 logs, respectively. Films with 3% and 1.5% cinnamaldehyde induced 1.2 to 2.8 and 1.2 to 1.3 log reductions, respectively. For  L. monocytogenes  on ham, carvacrol films induced greater reductions than did cinnamaldehyde films at all concentrations tested. In general, the reduction of  L. monocytogenes  on ham at 23 °C was greater than at 4 °C. Added antimicrobials had minor effects on physical properties of the films. The results suggest that the food industry and consumers could use these films as wrappings to control surface contamination by foodborne pathogenic microorganisms.     

Isolation and characterization of the microbial population of different South African kefir grains

Kefir, a slightly acidic fermented milk, is produced by adding lactic acid bacteria and yeasts, in the form of grains, to milk. The bacteria and yeasts present in the kefir grains are known to vary widely. Selective growth media and morphological and biochemical characteristics were used for the isolation and identification of the microbes present in the grains from eight different sources in South Africa. The kefir grains were activated in milk for only 24 h to prevent any changes in the microbial population of the grains. The microbial numbers varied between 6.4 × 10⁴ and 8.5 × 10 ⁸  cfu/g on the media selective for the bacterial species and between 1.5 × 10 ⁵ and 3.7 × 10 ⁸   cfu/g on the media selective for the yeast species. The bacterial genera that were identified included Lactobacillus , Leuconostoc and Lactococcus and the yeast genera included Zygosaccharomyces , Candida and Saccharomyces . The distribution frequencies of the microbes in the different grains were determined and most of the grains were dominated by two microbial species. No pediococci, acetic acid bacteria or propionibacteria were detected.     

侧向流动型重组酶聚合酶扩增技术检测食品中大肠埃希氏菌O157

研究内容基于侧向流动型重组酶聚合酶扩增(Lateral flow-based recombinase polymerase amplification,LF-RPA)技术原理,建立大肠埃希氏菌O157(Escherichia coli O157)的LF-RPA快速检测方法。应用该方法对3株大肠埃希氏菌O157和27株非大肠埃希氏菌O157进行检测,结果表明该方法具有良好的特异性。分别以10倍浓度梯度稀释插入靶基因序列的质粒pUC57-rfbE和大肠埃希氏菌O157菌液DNA作为模板进行LF-RPA检测。结果表明LF-RPA方法可检出1.0×10²拷贝/μL的pUC57-rfbE质粒和1.2×10⁴ cfu/mL的大肠埃希氏菌O157菌液。以大肠埃希氏菌O157污染食品样品,增菌4 h后,大肠埃希氏菌O157初始浓度为1.2×10¹ cfu/mL的食品增菌液可以被LF-RPA检出阳性信号。该研究建立的方法可快速、准确、灵敏地检测食品中大肠埃希氏菌O157,无需专业设备,在食品生产企业和基层检测机构具有一定的应用前景。     

Microbiological quality of ice creams commercialized in some cities in the state of Rio de Janeiro, Brazil

The microbiological quality of 60 ice cream samples of three commercial brands (A, B and C) of various flavours, commercialized in some towns in the State of Rio de Janeiro, Brazil, was evaluated. Total bacteria count (TCB), coliforms at 35°C (CT), coliforms at 44°C (CF) and presence of Staphylococcus aureus were performed on all samples. TCB ranged from 2.0 × 10 ² to 6.9 × 10 ⁵ cfu/mL, CT from < 3–≥ 2400 MPN/mL, CF from < 3–1100 MPN/mL and S. aureus from < 10–1.4 × 10 ⁶ cfu/mL. The level of bacterial contamination found in this study reflects the unhygienic conditions prevalent in manufacturing and storage of ice creams. Actions are thus necessary by the Brazilian regulatory agencies to require the ice cream processing plants to adopt quality guarantee systems, such as good manufacturing practices and hazard analysis and critical control points system.     

Postharvest Microbiology of Farm-reared, Tropical Freshwater Prawn (Macrobrachium Rosenbergii)

ABSTRACT: The microbiological quality of farm-reared, tropical freshwater prawn ( Macrobrachium rosenbergii ) stored at 2 different temperatures was studied. The prawn muscle was found to have the initial bacterial load of 10⁴ cfu/g. The lactics and vibrios were in the range of 10² cfu/g, while the E. coli , aeromonads, staphylococci, anaerobes, and molds were in the level of 10¹ cfu/g. Salmonella and Vibrio cholerae were present in the prawn muscle. The prawn muscle held at room temperature (28 ± 2 °C) was organoleptically acceptable up to 8 h, when the bacterial load was more than 10⁶ cfu/g. However, the prawn muscle stored at freezer temperatures (−10 to −15 °C) was found to be in acceptable condition even after 30 d of storage and the bacterial load was fluctuating in the range of 10³ to 10⁴ cfu/g.