ATP Bioluminescence Method Research Articles

Assessment of ATP-Bioluminescence and Dipslide Sampling to Determine the Efficacy of Slaughterhouse Cleaning and Disinfection Compared with Total Aerobic and Enterobacterales Counts

Inadequate cleaning and disinfection (C&D) in slaughterhouses can cause bacterial contamination of meat, resulting in foodborne disease and reduced meat quality. Different methods for monitoring the efficacy of C&D procedures are available, but few studies have assessed their reliability. This study examined C&D efficacy in slaughterhouses and evaluated the diagnostic performance of methods for measuring surface hygiene.One red meat and one poultry slaughterhouse in Sweden were each visited on six occasions before and six occasions after C&D. Sampling points were sampled with: swabbing and plating for total aerobic bacteria (TAB) and Enterobacterales (EB); dipslides for total viable count; and ATP-bioluminescence tests. To evaluate the diagnostic performance of the dipslide and ATP-bioluminescence methods, the results were compared with (TAB) as a reference.In total, 626 samples were collected. For the majority of samples, TAB was lower after than before C&D and EB were mainly detected before C&D, indicating C&D efficacy. Greater reductions in mean TAB were observed in processing areas (2.2 and 2.8 log CFU/100 cm2 in red meat and poultry slaughterhouse, respectively) than in slaughter areas (1.3 log CFU/100 cm2 in both slaughterhouses). Approximately half of all samples were assessed as non acceptably clean (52% for red meat and 46% for poultry slaughterhouse) according to previously published thresholds. Critical food contact surfaces that were insufficiently cleaned and disinfected were plucking fingers, shackles, and a post-dehairing table. Cleaning and disinfection of drains and floors were inadequate.The ATP-bioluminescence method showed low specificity compared with the reference (TAB) in both the red meat (0.30) and poultry slaughterhouses (0.64). The sensitivity of dipslides was low (0.26) in the red meat slaughterhouse compared with TAB. A combination of ATP-bioluminescence and dipslides could provide more accurate estimates of C&D efficacy.

ПРИМЕНЕНИЕ МЕТОДА АТФ-БИОЛЮМИНЕСЦЕНЦИИ ДЛЯ ОЦЕНКИ КАЧЕСТВА САНАЦИИ ДОИЛЬНОГО ОБОРУДОВАНИЯ

The article presents the results of determining the relationship between the total microbial number and microbial ATP on the surface of milking equipment before and after treatment with neutral anolyte ANK-SUPER. The possibility of using the ATP-bioluminescence method to control the quality of sanitation of milking equipment on dairy farms is shown. Laboratory studies revealed a proportional relationship between the level of bacterial ATP and the number of bacteria on the surface of milking equipment before and after sanitation.

Evaluation of ATP bioluminescence for monitoring surface hygiene in a hospital pharmacy cleanroom

The usefulness of the ATP bioluminescence method for monitoring surface hygiene was evaluated in a hospital pharmacy cleanroom. The sensitivity of the method was found to be appropriate for assessing the efficiency of cleaning and disinfection. ATP bioluminescence was superior to the traditional microbiological culture-based method for detecting unclean surfaces (p < .05).

Evaluation of different detector types in measurement of ATP bioluminescence compared to colony counting method for measuring bacterial burden of hospital surfaces

The ATP bioluminescence method has been increasingly employed as a rapid, on-site detection method in nosocomial infections control. In this study, we used a paired design of monitoring methods, the colony counting method (C) and the ATP bioluminescence method, to evaluate environmental surfaces after disinfection. The ATP bioluminescence method included three detector types (B, P, and N). Every surface after disinfection was performed by combining two types of monitoring methods or detectors. There was no statistically significant difference in theATP content per surface siteamong samples from intensive care units (ICUs)and internal medicine wards using B (p = 0.435) and P (p = 0.260). According to the Spearman’s rank correlation coefficients, with the exception of the correlation between the ATP content values detected by B and P, the correlation between the values generated by the remaining methods/detectors was weak or lacking, whereasthe differences between the detectors were statistically significant. Therefore, there are differences between the ATP bioluminescence method and the colony counting method, also between different detectors.

Modern Trends in Detection of Microbial Spoilage of Muscle Foods - A Review

Meat spoilage and detection is very important for meat technologists, quality control agencies, and the meat industry. Thus the microbial spoilage of meat and detection methods is widely studied. For decades microbial metabolites have been used as indicators of organoleptic spoilage of meat. Detection of spoilage by screening for microbial metabolites without identifying specific bacteria is the common approach. The ability to correlate biochemical change with microbial biomass is a complex problem. Current methods for the detection of spoilage in meats are inadequate, time consuming and labour intensive. ATP bioluminescence and fluorescent methods are already in use and are popular due to quick results. Molecular methods like multiplex PCR to detect a group of spoilage bacteria have already been tried. The concept of electronic nose using the odour sensors, conducting organic polymers and metal oxide conductors are promising method. The modified spectroscopy widely studied for detection of meat spoilage. Fourier transform infrared spectroscopy, electrical impedance spectroscopy, near infrared spectroscopy is some of those methods. The use of laser especially the laser speckle imaging is already put into use in this field. Latest technology introduced in the field of meat spoilage detection is the use of smart phone with different attachments. This paper briefly reviews the microbial spoilage of meat and microbial metabolites, then discusses about the currents methods for detection of spoilage and the modern upcoming methods with their potential.

EVALUATING THE PERFORMANCE OF ATP BIOLUMINESCENCE METHOD BY COMPARISON WITH CLASSICAL CULTURAL METHOD

Verification of the performance of the sanitation procedure is as important as the actual application. The methods that are going to be used for such verification needs to be user friendly and rapid resulting to apply the corrective actions promptly. Additionally, these methods should also be reliable besides their fast resulting. ATP bioluminescence method is recommended to be a solution for such procedures for many years. In this present study, our aim was to evaluate the performance of ATP bioluminescence method by comparison with golden standard classical cultural method. For this we performed parallel testing on surfaces which were experimentally contaminated with different types of microorganisms. The results showed that ATP bioluminescence gives more reliable results in surfaces which do not have any physical dirt and are contaminated with high level of microorganisms. The most reliable results were obtained from Salmonella contaminated surfaces.

Evaluation of an Amplified ATP Bioluminescence Method for Rapid Sterility Testing of Large Volume Parenteral

The sterility test described in pharmacopeial compendia requires a 14-day incubation period to obtain a valid analytical result. Therefore, the use of alternative methods to evaluate the sterility of pharmaceuticals, such as the Celsis AKuScreen™ Advance™ system, is particularly interesting because it allows a reduced incubation period and higher efficiency. The present study was aimed to evaluate and compare the performance of Celsis AKuScreen™ Advance™ system with the pharmacopeial sterility test. There was no significant difference between the ability of detection of microbial contamination observed within pharmacopeial method and test method. The Celsis AKuScreen™ Advance™ system allowed a faster detection of the challenge microorganisms, which indicates that the system is a viable alternative for assessing the sterility of injectable products.

Study on the Detection of Total Colony in Medical and Health Environment Based on ATP Bioluminescence

In this paper, the application of ATP fluorescence in the detection of colonies in the health environment of hospitals was studied. Firstly, the principle of ATP bioluminescence method was described. Then, ATP bioluminescence and plate count method were used to test the density of the surface of the objects in selected area, taking the time points 2 hours after disinfection as the time nodes. The results showed that the difference between the qualified rate of ATP bioluminescence assay and the plate count method was statistically significant {P&lt;0.01}. Therefore, ATP bioluminescence method was highly correlated with bacterial culture method. The correlation coefficient of pass rate of the two methods was 0.782, which indicated that there was a positive correlation between the two test results. Besides, the detection results showed that ATP bioluminescence method had higher sensitivity than plate counting method. Therefore, ATP bioluminescence method was more suitable for the rapid detection of the colony of hospital health environment, and helps the hospital to better manage its environmental hygiene conditions.

ATP-Bioluminescence and Conventional Microbiology for Hygiene Evaluation of Cutting Room Surfaces in Poultry Slaughterhouse

Background: The hygiene procedures in poultry slaughterhouses consist in the use of hot water, detergent and sanitizing, configuring Sanitation Standard Operating Procedure (SSOP). These actions control contamination in food processing environments, especially by pathogenic microorganisms, which cause diseases with impact on public health and economic losses. The microbiological control of aerobic mesophiles, Staphylococcus aureus and Escherichia coli, are used as indica­tors of contamination. The hygienic-sanitary conditions on the surfaces of the poultry slaughterhouse cuttting room were evaluated, before and after cleaning and sanitizing procedures.Materials, Methods &amp; Results: Conventional microbiology (Rodac plates and sponge for quantification of aerobic me­sophiles, Staphylococcus aureus and Escherichia coli) and ATP-Bioluminescence were used to analyze the action of hot water and the active principles peracetic acid, quaternary ammonia and biguanide in the standard pre-operational hygiene procedure in the cutting room of the poultry slaughterhouse under Federal Inspection with slaughter capacity of more than 20.000 birds/h. The evaluations were performed on three lines of chicken thigh cuts at the same time and in a completely randomized manner on stainless steel surfaces, polyurethane belts and polyethylene boards. Samples were made in four replicates at the three surface totaling 108 assay for each microorganism. The samples were collected at the end of the cut­ting process, before and after washing the surfaces with hot water (between 45 and 50ºC) and after sanitization with 0.5% peracetic acid, 2% quaternary ammonia and 1% biguanide. The ATP-Bioluminescence method detected organic matter at all collected points and Rodac plates allowed a better recovery of microorganisms than sponges for quantification of aerobic mesophiles, E. coli and S. aureus. There was a reduction of contamination after the action of hot water and, after using quaternary ammonia and peracetic acid, there was no isolation of E. coli and S. aureus on all evaluated surfaces.Discussion: The use of different methods of analysis for monitoring the hygiene and sanitary status of contact surfaces with chicken cuts allows greater flexibility in relation to hygiene control. The use of the bioluminescent ATP detection method allows detecting in seconds extremely low levels of contamination, allowing a quick determination of the cleaning efficiency on the surfaces and evaluation of the hygiene programs. Conventional microbiology methods, on the other hand, provide indicators of contamination by different microorganisms on food contact surfaces. Both are applicable in SSOP monitor­ing programs and sanitary conditions of the contact surfaces in food producing establishments. The significant reduction of microorganisms on surfaces after cleaning, found in this study, demonstrates the importance of operational hygiene in the maintenance of microbial contamination below the recommended limits, and to reconcile the ATP-Bioluminescence methodologies and Rodac plates can bring benefits to the control of this contamination, and the use of ATP-bioluminescence makes possible taking immediate corrective measures after the evaluation of sanitation procedures.Keywords: ATP-Bioluminescence, Rodac plates, Escherichia coli, mesophiles, Staphylococcus aureus.

ATP-Bioluminescence and Conventional Microbiology for Hygiene Evaluation of Cutting Room Surfaces in Poultry Slaughterhouse.

Background: The hygiene procedures in poultry slaughterhouses consist in the use of hot water, detergent and sanitizing, configuring Sanitation Standard Operating Procedure (SSOP). These actions control contamination in food processing environments, especially by pathogenic microorganisms, which cause diseases with impact on public health and economic losses. The microbiological control of aerobic mesophiles, Staphylococcus aureus and Escherichia coli, are used as indicators of contamination. The hygienic-sanitary conditions on the surfaces of the poultry slaughterhouse cuttting room were evaluated, before and after cleaning and sanitizing procedures.Materials, Methods &amp; Results: Conventional microbiology (Rodac plates and sponge for quantification of aerobic mesophiles, Staphylococcus aureus and Escherichia coli) and ATP-Bioluminescence were used to analyze the action of hot water and the active principles peracetic acid, quaternary ammonia and biguanide in the standard pre-operational hygiene procedure in the cutting room of the poultry slaughterhouse under Federal Inspection with slaughter capacity of more than 20.000 birds/h. The evaluations were performed on three lines of chicken thigh cuts at the same time and in a completely randomized manner on stainless steel surfaces, polyurethane belts and polyethylene boards. Samples were made in four replicates at the three surface totaling 108 assay for each microorganism. The samples were collected at the end of the cuttingprocess, before and after washing the surfaces with hot water (between 45 and 50ºC) and after sanitization with 0.5% peracetic acid, 2% quaternary ammonia and 1% biguanide. The ATP-Bioluminescence method detected organic matter at all collected points and Rodac plates allowed a better recovery of microorganisms than sponges for quantification of aerobic mesophiles, E. coli and S. aureus. There was a reduction of contamination after the action of hot water and, after using quaternary ammonia and peracetic acid, there was no isolation of E. coli and S. aureus on all evaluated surfaces.Discussion: The use of different methods of analysis for monitoring the hygiene and sanitary status of contact surfaces with chicken cuts allows greater flexibility in relation to hygiene control. The use of the bioluminescent ATP detection method allows detecting in seconds extremely low levels of contamination, allowing a quick determination of the cleaning efficiency on the surfaces and evaluation of the hygiene programs. Conventional microbiology methods, on the other hand, provideindicators of contamination by different microorganisms on food contact surfaces. Both are applicable in SSOP monitoring programs and sanitary conditions of the contact surfaces in food producing establishments. The significant reduction of microorganisms on surfaces after cleaning, found in this study, demonstrates the importance of operational hygiene inthe maintenance of microbial contamination below the recommended limits, and to reconcile the ATP-Bioluminescence methodologies and Rodac plates can bring benefits to the control of this contamination, and the use of ATP-bioluminescence makes possible taking immediate corrective measures after the evaluation of sanitation procedures.Keywords: ATP-Bioluminescence, Rodac plates, Escherichia coli, mesophiles, Staphylococcus aureus.

Microbial load of white cheese process lines after CIP and COP: A case study in Turkey

Microbial adhesion, contamination and biofilm formation are major problems in the dairy industry, in cheese production, and for consumer health. In our study, the microbial loads of white cheese process lines were examined after cleaning treatments. Bacillus spp. was determined as the most common species on process lines. Drainage, air, and water entry points were found as important contamination points. Biofilm forming capacity of Enterobacteriaceae as revealed using Congo Red agar, was very high. In addition, the standard counting and ATP-Bioluminescence methods were applied and compared for evaluating biofilms. The ATP-Bioluminescence method was found useful for rapidly identifying the biofilm-formable surfaces.

Comparative evaluation of baseline cleanliness of food contact surfaces in fishery based microenterprise units by ATP-Bioluminescence and conventional microbiological methods

The baseline cleanliness of food contact surfaces namely floor, working tables, workers’ hands, utensils and machinery of a fishery-based microenterprise were examined by traditional microbiological swab analysis method and ATP-bioluminescence method. The study showed that swab results, measured in relative light unit which is proportional to total recovered ATP, did not have significant correlation with standard method of microbiological swab in the adjacent sites of equal area. The study also examined the bactericidal efficiency of selected commercially available sanitisers and the effect of treatment was found to be significantly different from each other. Analysis of percentage reduction of bacterial load on food contact surfaces showed that commercially available stabilised hydrogen peroxide was more effective than sodium hypochlorite used in the food industries (p&lt;0.05).

ATP bioluminescence method: tool for rapid screening of organic and microbial contaminants on deteriorated mural paintings

The extent of the microbial contamination of the seventeenth-century wall paintings in the nave of the old Church of the Holy Ascension (Veliki Krčimir, Serbia) was evaluated via newly implemented ATP bioluminescence method, and traditional cultivation-based method, utilising commercially available dip slides. To assess the validity of ATP, as a biomarker for rapid detection of mural surface contamination, obtained zones of cleanliness values, in range from 1.0 to 5.3, were compared to documented total microbial counts, ranging between seven and 247 CFU/cm2. Small coefficients of determination, 0.0106–0.0385, suggest poor correlation between microbial counts and surface ATP levels; however, zones of cleanliness values are of great help in determining the high points of contamination, aka ‘hotspots’, which should be given special attention during sampling and investigation using other methods. In addition, various aspects of the possible implementation of the ATP bioluminescence method in an integrated system of wall painting conservation are discussed.

Mathematical modeling and cycle time reduction of deposit removal from stainless steel pipeline during cleaning-in-place of milking system with electrolyzed oxidizing water

The safety of raw milk largely depends on using a clean milking system during the milk production. The milking system cleaning process widely used on dairy farms is a highly automated process called cleaning-in-place (CIP), which comprises of four cycles: i) warm water rinse; ii) alkaline wash; iii) acid wash; and iv) sanitizing rinse before the next milking event. Electrolyzed oxidizing (EO) water is an emerging technology, which consists of acidic and alkaline solutions by the electrodialysis of dilute sodium chloride solution. Previous studies in our lab showed that EO water can be an alternative for milking system CIP. Despite the progress made to enhance the CIP performance and evaluate alternative CIPs, the mechanisms behind the cleaning processes were still largely unclear. Therefore, this study was undertaken to evaluate the deposit removal rate during the EO water CIP process using a stainless steel surface evaluation simulator. Deposit removal data from the simulator formed the basis for developing mathematical models to describe the deposit removal process during the CIP process with EO water. Stainless steel straight pipe specimens were placed at the end of undisturbed entrance length along the simulator pipeline. The mass of milk deposits on the inner surfaces of the specimens were measured using a high precision balance after the initial soiling, and then after certain time durations within the warm water rinse, alkaline wash, and acid wash cycles. A unified first order deposit removal rate model dependent on nth power of remaining deposit mass was used for all three cycles. ATP bioluminescence method was also used as a validation approach at the end of each CIP cycle. Experimental results showed that the milk deposit on the inner surfaces of the specimens was removed rapidly by the warm water rinse within 10 s of rinse time. For the alkaline and acid wash cycles, the co-existence of a fast deposit removal at the beginning of the wash cycle and a slow deposit removal throughout the entire wash cycle was inferred. The proposed models matched the experimental data with small root mean square errors (0.23 mg/[(mg) (m2)] and 0.07 mg/[(mg) (m2)] for the upstream and downstream locations, respectively) and satisfactory percent error differences (3.67% and 0.93% for the upstream and downstream locations, respectively). Based on the experimental data and the proposed models, the time duration of the CIP process was shortened by 55% (10 s warm water rinse, 3 min alkaline wash and 6 min acid wash) and validated, which yielded an average deposit of 0.28 mg/[(mg) (m2)] at the end of the CIP as compared with that of 0.29 mg/[(mg) (m2)] at the end of the original CIP, to achieve a satisfactory CIP performance for the simulator.

Urine analysis test paper in the application of equipment cleaning quality evaluation

Objective To observe the urine analysis test paper evaluation method for the determination of the sensitivity of medical equipment cleaning effect, for cleaning quality evaluation method. Methods To perform standard manual processing equipment in the experiments, the first use of ATP monitoring method and the analysis of urine test paper test mid experiment data, calculate its whether there is relationship and ATP test method, and can instruct the cleaning quality results; Second use visual method, ATP bioluminescence method and the analysis of urine test paper test the end of the experiment data, the analysis to determine the urine test paper measurement indicator cleaning qualified evaluation threshold. Results The ATP test paper method and urine analysis had showed positive correlation, 0.30≤r≤0.75, and to complete the standard cleaning process 300 pieces of department of gynaecology attractor visual analysis method, ATP bioluminescence method and the analysis of urine test paper measurement inspection group (the -), cleaning the percent of pass was 95.67%(287/300), 86.33%(259/300) and 83.37%(251/300) respectively, therefore, urine analysis test paper assay to detect the - was the cleaning threshold quality qualification evaluation, urine analysis test paper test the percent of pass was 14.46% lower than the visual observation, χ2= 63.35, P<0.05. Conclusions Urine analysis test paper test results, test method is simple, low cost, and can be widely used in medical equipment cleaning quality evaluation method of semi-quantitative daily monitoring. Key words: Urinalysis; Medical device; Cleaning effect; Evaluation method

A two-hour antibiotic susceptibility test by ATP-bioluminescence

The antibiotic susceptibility test (AST) in Clinical Microbiology laboratories is still time-consuming, and most procedures take 24h to yield results. In this study, a rapid antimicrobial susceptibility test using ATP-bioluminescence has been developed. The design of method was performed using five ATCC collection strains of known susceptibility. This procedure was then validated against standard commercial methods on 10 strains of enterococci, 10 staphylococci, 10 non-fermenting gram negative bacilli, and 13 Enterobacteriaceae from patients. The agreement obtained in the sensitivity between the ATP-bioluminescence method and commercial methods (E-test, MicroScan and VITEK2) was 100%. In summary, the preliminary results obtained in this work show that the ATP-bioluminescence method could provide a fast and reliable AST in two hours.

Evaluation of ATP Bioluminescence method to monitor the quality of reprocessing flexible endoscopes

Evaluation of ATP Bioluminescence method to monitor the quality of reprocessing flexible endoscopes

Effects of process stages on Turkish delight/lokum microbiological quality

SummaryThe sources of microbiological contamination at the lokum production lines of two local confectionaries were determined. Surface samples from the production lines were evaluated by standard and ATP bioluminescence methods. Microbial loads of raw materials were also determined. The trays and cutting knives were found to be the primary contamination sources, and personnel hands were found to be the secondary contamination sources, while all of them were found to be the secondary sources when the standard swab method was applied. Raw materials were not sources of contamination, because of their microbiological loads and the following high‐temperature cooking stages. The microbial loads of lokum also were not high, due to product contents and the processes applied to lokum. There is a need for further studies related to the effects of process steps on the physical and the chemical quality of lokum. Training for workers in food safety at the confectionaries is also needed.

Study on ATP bioluminescence in assaying the cleaning quality of uterine suction

Objective To explore the application value of ATP bioluminescence in assaying the cleaning quality of uterine suction.Methods According to the principle of random,we selected 120 uterine suction tube after use.60 used tubes in A group were given water immersion + high-pressure water jets wash + dry method,60 used tubes in B group were given alkaline cleaning solution soak + ultrasonic + high-pressure water washing + manual scrub + drying method.The quality of suction tube was initially tested with the visual method after cleaning,Straw inside the top cavity,2.5cm at inside straw joint and the front of straw outer wall,10cm at suction cavity wall,before and after cleaning the residues of bacteria were assayed with the ATP bioluminescence and bacteria counting method for parallel comparison.Results After cleaning drying program,the RLU of two groups in suction head internal top,2.5cm at inside straw joint,the front of straw outer wall,10cm at suction cavity wall had significant differences (t =26.81,29.13,7.14,55.74,P ＜ 0.01).The cleaning effect of B group was better than A group..Bacteria counting method was used to detect pathogenic microorganism that was not detected.The qualified rate of cleaning at four parts was as following:group A:70.0％,85.0％,93.3％,93.3％.B group:95.0％,98.3％,100.0％,100.0％,and the qualified rate of group B was significantly higher than group A (x2 =12.99,6.98,4.14,4.14,all P ＜ 0.05).Conclusion ATP bioluminescence method can dynamically rapid detect the bacteria before and after intrauterine suction cleaning residues,can be viewed as an effective method to detect quality of uterine suction cleaning. Key words: Luciferases, bacteria; Surgical instruments; Quality control

Estimation of total bacteria on mango surface by using ATP bioluminescence

This paper presents a rapid ATP assay for monitoring the total bacterial load associated with mango fruit surface. Conventional methods such as total plate count for the determination of total microbial counts are time consuming and prone to contamination resulting in low accuracy. Thus the need for quick and accurate methods for determination of total microbial count has led to development of many rapid techniques such as ATP bioluminescence. In this study the potential of ATP bioluminescence method for the determination of total bacterial count on mango surface has been evaluated using two types of luminescence techniques based on fluorescence spectroscopy and bioluminescence. Various mechanical and chemical extraction processes were employed for the extraction of ATP from bacterial cells and the best extraction was achieved with 0.03M TCA. Minimum level of detection of ATP using fluorescence spectrophotometer and bioluminometer was found to be 10−8M and 10−13M, respectively. This ATP concentration was correlated with the number of bacterial cells present on mango surface. The minimum level of detection of bacterial cells in fluorescence spectroscopy and bioluminescence was found to be 107cfu/ml and 102cfu/ml, respectively.