Research note: A scald water surfactant combined with an organic acid carcass dip reduces microbial contaminants on broiler carcasses during processing.

Organic acids are applied to poultry carcasses during processing to reduce foodborne pathogens and spoilage microorganisms. Scald water surfactant agents employed to improve feather removal may enhance the efficacy of organic acids during processing. This study investigated the effects of concurrent application of a scald water surfactant and organic acid dip on microbial contamination of carcasses processed in a small-scale production model. Broilers were reared in litter floor pens to 47 d of age and slaughtered using standard practices. Carcasses were scalded in either control or surfactant scald water initially and dipped in either a 2% organic acid blend or water after feather removal to complete a 2 × 2 factorial arrangement with n = 15 carcasses per treatment group. The commercially available scald water additive was a slightly alkaline surfactant solution labelled as a feather removal aid. The organic acid dip consisting of lactic and citric acid was maintained at pH of 2.5. Approximately 10 g of neck skin was collected 1-min postdipping and placed in buffered peptone water with an added neutralizing agent, sodium thiosulfate. Serial dilutions were performed to determine general coliform (GC), E. coli (EC), and aerobic plate (APC) counts as CFU per gram of skin sample. A significant 0.61, 0.76, and 1.6 log reduction of GC, EC, and APC, respectively, was attributed to use of the organic acid carcass dip (P ≤ 0.01). There were no significant differences in carcass microbial reduction due to surfactant scald water alone. A 0.69, 0.73 (P ≤ 0.05), and 1.96 log reduction of GC, EC, and APC, respectively, was observed in surfactant-scalded, acid-dipped carcasses compared to water-scalded, water-dipped control groups. These data demonstrated that a surfactant scald water additive and an organic acid carcass dip can have beneficial effects of microbial reduction when employed simultaneously during broiler processing.

Isolation of Shiga Toxin-Producing Escherichia coli from the Surfaces of Beef Carcasses in Slaughterhouses in Japan.

Shiga toxin-producing E. coli (STEC) is an important foodborne pathogen worldwide. It is necessary to control and prevent STEC contamination on beef carcasses in slaughterhouses because STEC infection is associated with beef consumption. However, the frequencies of STEC contamination of beef carcasses in various slaughterhouses in Japan are not well known. Herein, we investigated the contamination of beef carcasses with STEC in slaughterhouses to assess the potential risks of STEC. In total, 524 gauze samples were collected from the surfaces of beef carcasses at 12 domestic slaughterhouses from November 2020 to February 2023. The samples were measured for aerobic plate counts and tested for pathogenic genes (stx and eae) and major O-serogroups (O26, O45, O103, O111, O121, O145, and O157) by real-time PCR screening. Subsequently, immunomagnetic separation (IMS) was performed on samples positive for stx, eae, and at least one of the seven O-serogroups of STEC. Isolation process without IMS was performed on samples positive for stx, including those subjected to IMS. STEC O157:H7 and stx-positive E. coli other than serotype O157:H7 were isolated from 0.6% and 4.6% of beef carcass surfaces, respectively. Although the STEC O157:H7 isolation rate was low and stx-positive E. coli other than serotype O157:H7 belonged to minor O-serogroups, the results mean a risk of foodborne illness. Furthermore, a moderate correlation was observed between aerobic plate counts and detection rates of stx-positive samples by real-time PCR screening. The STEC O157:H7 isolated facilities showed higher values on aerobic plate counts and detection rates of stx-positive samples than the mean values of total samples. Therefore, these results suggest that it is important to evaluate hygiene treatments against beef carcasses for the reduction of STEC contamination risk, particularly in facilities with high aerobic plate counts.

Research Note: Reducing food loss in the manufacturing process of chickens by reconditioning dropped raw poultry carcasses with peroxyacetic acid and sodium hypochlorite (chlorine) solution.

Food waste and food loss has been a growing concern in the manufacturing industry with a gap between identifying the problem and implementing a solution. The manufacturing process of chicken is largely automated by conveyor belts and machines in which initial application of either peroxyacetic acid (PAA) or sodium hypochlorite (chlorine) solution is utilized to reduce the microbial load and prevent food borne illnesses on the chicken products as they are processed and packaged for distribution. However, during this automated process whole chickens can drop from the manufacturing line and become contaminated leading to the disposal and waste of the product. A solution to reduce food waste was to analyze a reconditioning procedure within the manufacturing process. The study evaluated the aerobic microbial growth on salvaged marinated deli raw whole chickens without giblets (WOGs) from conveyor belt loss reconditioned in either PAA or sodium hypochlorite (chlorine) solution to undropped chicken WOGs. Chicken rinsate and segmented samples were collected from each parameter and tested for microbial growth using Petrifilm aerobic plate count (APC) plates and converting results into log colony forming units (CFU). A difference (P < 0.05) was observed with the reconditioning of the WOGs in PAA (0.71 log10 CFU/mL) compared to the control (1.45 ± 0.26 log10 CFU/mL), for rinses. Of the segmented samples, the trussing strings displayed a significant decrease in APC counts for both chlorine (2.30 ± 0.49 log10 CFU/g) and PAA (2.3 ± 0.49 log10 CFU/g) reconditioning compared to the control (2.72 ± 0.39 log10 CFU/g). Reconditioning of salvaged deli chicken WOGs in chlorine or PAA is comparable to or better than the conventional process for the reduction of APC, it is an effective strategy to reintroduce dropped marinated deli chicken WOGs to the manufacturing line and can reduce food waste at a manufacturing level.

The Influence of Storage Temperature and Packaging Technology on the Durability of Ready-to-Eat Preservative-Free Meat Bars with Dried Plasma.

Food business operators must include the results of shelf life testing in their HACCP plan. Ready-to-eat preservative-free meat products enriched with blood plasma are an unfathomable area of research in food safety. We tested modified atmosphere (80% N2 and 20% CO2) and vacuum packaged RTE preservative-free baked and smoked pork bars with dried blood plasma for Aerobic Plate Count, yeast and mould, lactic acid bacteria, Staphylococcus aureus, Enterobacteriaceae, Escherichia coli, and Campylobacter spp., and the presence of Listeria monocytogenes and Salmonella spp. during storage (temperatures from 4 to 34 °C) up to 35 days after production. The obtained data on the count of individual groups of microorganisms were subjected to analysis of variance (ANOVA) and statistically tested (Student's t-test with the Bonferroni correction); for temperatures at which there were statistically significant differences and high numerical variability, the trend of changes in bacterial counts were visualised using mathematical modelling. The results show that the optimal storage conditions are refrigerated temperatures (up to 8 °C) for two weeks. At higher temperatures, food spoilage occurred due to the growth of aerobic bacteria, lactic acid bacteria, yeast, and mould. The MAP packaging method was more conducive to spoilage of the bars, especially in temperatures over 8 °C.

[Microbial contamination in dried fruit products in China in 2019].

OBJECTIVE: To investigate the microbial contamination in dried fruit products in China. METHODS: In 2019, 2917 samples of dried fruit products on the market were collected, and examined for aerobic bacterial count, coliforms, molds, yeasts, Salmonella and Listeria monocytogenes according to the method specified in GB 4789. RESULTS: A total of 34.42%(1004/2917)of the samples had molds above 50 CFU/g and 9.46%(276/2917)of the samples had yeast above 50 CFU/g. The occurrence of aerobic plate count above 10~4 CFU/g and coliforms above 10~2 CFU/g was 5.01%(146/2917)and 2.98%(87/2917), respectively. The detection rate of Salmonella and Listeria monocytogenes were 0.14%(4/2917) and 0.03%(1/2917), respectively. Microbial contamination in different kinds of dried fruit products varied widely, with dried wolfberries and dried durian having the worst overall hygiene. There were differences in microbial contamination of dried fruit products in different regions. In general, samples collected in South China, Southwest China and Central China had more serious microbial contamination. There was no significant difference in microbial contamination between dried fruit products with different packaging and sampling places. CONCLUSION: The hygienic condition of dried fruit products is generally poor in 2019.

Incubation temperature and culture medium formulation impact the accuracy of pour-plate techniques for the enumeration of industrial Bacillus assemblages.

Aerobic plate counting assays based on the pour-plate technique are frequently used to enumerate microbial products; however, colony swarming and merging at the agar surface can reduce the accuracy of these assays. Some plating methods mitigate this risk through the inclusion of strategies including agar overlays; however, these interventions may be inadequate to mitigate swarming and merging of certain Bacillus colonies. In the present study, we assessed the accuracy of several pour-plate techniques for the enumeration of a mixed-species Bacillus assemblage. Tested modifications included a customized culture medium formulation, agar overlays, decreased incubation times and increased incubation temperature. Methods which produced countable plates were assessed for agreement with a Bacillus-specific plate counting assay and with total cell counts rendered by flow cytometry. While all tested pour-plate methods underestimated Bacillus endospore concentrations relative to flow cytometry and customized spread-plating, our results suggest that increasing incubation temperature and the inclusion of bile salts into culture medium formulations can improve the accuracy of pour-plate techniques when used to enumerate Bacillus assemblages by decreasing the incidence of spreading colonies. As Bacillus endospore preparations become more ubiquitous in the market, familiar enumeration methods such as the pour-plate technique may require methodological modifications to ensure that the cGMP compliance of Bacillus-based microbial products is assessed accurately.

Rapid detection of total bacteria in foods using a poly-l-lysine-based lateral-flow assay.

Food safety and freshness are evaluated according to microbiological load. To analyze this load rapidly, a poly-l-lysine-based lateral-flow assay (PLFA) was developed. A total of 90 strains of bacteria that are often detected in spoiled foods, including Enterobacteriaceae, lactic acid bacteria, Pseudomonas, and Bacillus were detected using the PLFA. A positive signal was obtained when the bacterial concentration was ≥6 log10 (cfu/test). A total of 36 fresh foods (meats, pastries, lettuces, cabbages, radishes, and sprouts) and corresponding spoiled foods were cultured for 0, 3, 6, and 9 h to investigate how many hours were required for microbial detection using PLFA. The higher the number of bacteria in a food, the shorter was the culture time required for PLFA-positive results to be obtained, so the distinction between fresh and spoiled food could be made based on the time taken for the culture to become PLFA-positive. The coefficient of determination of the least squares regression between the time to become PLFA-positive and the initial log10 (cfu/g) bacterial count for the food was 0.9888. The test time for the PLFA, including pretreatment, was approximately 15-30 min. This novel method will enable the detection of total bacteria on the food processing site.

The Microbial Quality and Safety of Blenderised Enteral Nutrition Formula: A Systematic Review.

The use of blenderised enteral nutrition formula (ENF) is on the increase globally. However, concerns remain regarding the microbial quality and safety of blenderised ENF compared with standard recommendations and commercial ENF. AIM: This was a systematic review which sought to compare the microbial quality of blenderised ENF and commercial ENF and to evaluate the effect of storage time on blenderised ENF. METHOD: Four databases (Pubmed, EMBASE, PSYCInfo and Google scholar) were searched for relevant articles based on the Population, Intervention, Comparator, Outcomes framework. RESULTS: Eleven studies which met the criteria were included in the systematic review. Two major areas were identified; Microbial Quality of Blenderised ENF versus Commercial ENF; and The Effect of Storage Time on Microbial Quality of Blenderised ENF. Overall, 72.7% of the studies showed microbial contamination in blenderised ENF compared with 57.1% of commercial ENF, and the storage time was another important factor in the rates of contamination. The extent of handling or manipulation of the enteral formula was critical in determining the level of contamination. CONCLUSION: Preparation techniques for blenderised ENF need to be established and caregivers taught how to prepare and administer it appropriately in order to reduce contamination. Further, well-designed studies are required, which compare the microbial quality of blenderised ENF using adequate handling techniques and commercial ENF.

Culture medium brand choice impacts colony swarming behavior among industrial Bacillus isolates and the accuracy of aerobic plate counts.

Aerobic plate count assays are an industry standard method for the enumeration of microbial products. Colony swarming among industrial Bacillus isolates on solid medium can impact a counting technician's interpretation of colony count, promote inter-technician variance and reduce the agreement of plate counts with growth-independent enumeration methods. In the present study, we examined swarming behavior among four industrial Bacillus species as a function of culture medium brand choice. Colony diameter for three Bacillus species was found to be influenced by culture medium brand, as was colony count interpretation among three out of four plate counting technicians. Estimations of Bacillus endospore concentration were likewise influenced by culture medium brand, leading to an increased incidence of QC failure for replicate samples of a Bacillus-based microbial product as a function of brand choice. Results suggest that culture medium brand choice may be an additional source of variance when plate counting is used for the enumeration of Bacillus-based microbial products. We recommend that plating medium brand availability in testing laboratories be considered by microbial product manufacturers when considering sources of variance in customer and regulatory laboratories.

Enumeration of industrial Bacillus assemblages in commercial products with customized plate-counting assays.

The aerobic plate count assay remains among the most widespread methods for enumerating industrial Bacillus assemblages, as growth-independent methods are either cost prohibitive or unavailable in some areas. However, the standard plating assays used to verify the CFU count of Bacillus-based products are not tailored to Bacillus species and thus may not produce the most accurate possible estimations. Standard plating assays assume that established limits of quantification are applicable to Bacillus species whose colonies swarm on solid media, and that colonies of each species in a mixed-species assemblage form independently of one another on agar plates. In the present study, we examined the upper limit of quantification for an assemblage of swarming industrial Bacillus isolates by comparing plate count on medium with and without a swarming inhibitor with direct counts for spore suspensions of increasing endospore concentration. Additionally, we examined the impact of assemblage species composition on the evenness of colony distribution across replicate plates for four industrial Bacillus isolates. We compared the observed distribution of colonies across replicate plates to the expected Poisson distribution for axenic endospore suspensions, for a 3-species assemblage and a 4-species assemblage, respectively. Results suggest that customized plating assays may be more appropriate than standard protocols for the enumeration of Bacillus-based products, and that interactions between colonies on solid media should be considered when interpreting plating data for mixed-species Bacillus assemblages.

Microbiological Testing Results of Cooked Diced Chicken and Pasteurized Egg Products Purchased for United States Federal Nutrition Assistance Programs, 2012-2018.

The Agricultural Marketing Service (AMS) purchases cooked diced chicken, pasteurized liquid whole eggs, and pasteurized dried egg mix for federal nutrition assistance programs. Purchases are made from establishments that have met the financial and technical requirements to become AMS-approved vendors. Cooked diced chicken is tested for the presence of Salmonella and Listeria monocytogenes, and for levels of aerobic plate count (APC) organisms, coliforms, and generic Escherichia coli (GEC). Of 3668 samples collected from October 2012 through September 2018, none were positive for Salmonella, 3 (0.8%) were positive for L. monocytogenes, 8 (0.22%) exceeded the APC critical limit (CL) of 1000 colony-forming units (CFUs)/mL, 15 (0.41%) exceeded the coliform CL of 50 CFU/mL, and 5 (0.14%) exceeded the GEC CL of 10 CFU/mL. Pasteurized liquid whole egg and pasteurized dried egg mix are tested for the presence of Salmonella and for levels of APC and CL. Of 984 pasteurized liquid whole egg samples collected from October 2012 through September 2018, 1 (0.10%) was positive for Salmonella, 29 (2.5%) exceeded the APC CL, and 4 (0.41%) exceeded the coliform CL. Of 380 pasteurized dried egg mix samples collected during this period, none was positive for Salmonella, none exceeded the APC CL, and 3 (0.79%) exceeded the coliform CL. All production lots from which samples found to contain pathogens or to exceed indicator organism CLs were identified and rejected for purchase by AMS. These data suggest that cooked diced chicken and pasteurized egg products produced for federal nutrition assistance programs are done so under effective food safety systems.

Comparison of electronic sensing techniques for screening dried shrimps irradiated using three types of approved radiation with standard analytical methods.

Rapid analytical methods for screening irradiated foods are required to comply with the approved standards for international trade. Dried shrimps irradiated at 1-7 kGy with gamma rays, electron beam (E-beam), and X-rays were screened with an electronic nose (E-nose) and electronic tongue (E-tongue). The data were compared with those from European standard methods (photostimulated luminescence, PSL) and direct epifluorescent filter technique/aerobic plate count, DEFT/APC). All irradiated shrimp samples were clearly discriminated from the non-irradiated control based on PSL photon count measurements and DEFT/APC microbial enumeration. The volatile patterns and taste attributes of the irradiated (>1 kGy from three sources) and control samples could be distinguished by the E-nose and E-tongue analyses through principal component analysis. Verification through electron spin resonance and thermoluminescence analyses validated screening results. The results indicate that E-sensing techniques showed potential for the rapid screening of irradiated foods like dried shrimps.

Disinfectant of pummelo (Citrus Grandis L. Osbeck) fruit juice using gaseous ozone.

This work studied the effectiveness of gaseous ozone disinfection on pummelo (Citrus Grandis L. Osbeck) fruit juice components. Unfiltered and filtered pummelo fruit juices were treated with gaseous ozone for up to 50 min with ozone concentration fixed at 600 mg/h. A microbiological and physicochemical properties analysis were conducted on the ozone-treated fruit juices samples. It was found that the survival rate of aerobic bacteria, yeast and mold in unfiltered pummelo fruit juice were higher compared to filtered juice, as the juice components acted as protective barriers to the microorganisms. The microorganisms' inactivation in pummelo fruit juices was also observed to have increased as the ozone treatment time increased. Significant effects on total colour difference, ascorbic acid content, and total phenolic content were also observed over increased ozone-treatment time. However, ozone was shown to be ineffective in activating PME activity in both types of juice. The experimental results of this study indicated that pummelo fruit juice components had significant effects on the effectiveness of gaseous ozone, however, the degree of the effects depends on the different fruit components (total soluble solids, total phenolic content). As a conclusion, filtered juice showed better quality characteristics in comparison to unfiltered juice post-ozone treatment.

Evaluation of the Microbiological Status of Raw Beef in Korea: Considering the Suitability of Aerobic Plate Count Guidelines.

This study was conducted to analyze the microbiological contamination status of raw beef distributed in Korea, and evaluate the suitability of current aerobic plate count (APC) guidelines. We analyzed five years (2010-2014) of microbiological monitoring data obtained from the Ministry of Food and Drug Safety and investigated the microbiological status of raw beef collected from meat packing centers and meat shops in the Seoul/Gyeonggi, Gangwon, and Chungcheong regions in August 2015. From 2010-2014, most raw beef (>94%) displayed APC levels of < 1.0 × 106 CFU/g. However, raw beef samples collected from all three regions in August 2015 had comparatively higher APC levels than those reported in previous years. To evaluate the relationship between the APC level and quality, changes in beef loin were evaluated during cold storage for 15 days at 4°C. On day 11, the mean APC level (4.7 × 106 CFU/g) conformed to current guidelines in Korea (1.0 × 107 CFU/g) and the pH value was 5.82. However, the sensory evaluation score for color and overall acceptability was under 3.0, meaning that the beef loin was not acceptable for eating. These results suggest that current APC guideline for raw beef should be lowered to 1.0 × 106 CFU/g to improve both the microbiological safety and palatability of raw beef.

Microbiological Quality and Incidence of Salmonella on Cherry Tomatoes at Retail in Querétaro, México.

Multiple outbreaks related to Salmonella in tomatoes require an evaluation of the risk associated with cherry tomatoes due to the increase in its production, consumption, and marketing in Mexico's central region. The purpose of this study was to determine the microbial quality of cherry tomatoes obtained from two retail sale points (supermarkets and local markets). Cherry tomato samples (333) were collected from four supermarkets and from four local markets, and the contents of aerobic plate count, molds and yeasts, total coliforms, and Escherichia coli were quantified; the presence of Salmonella was simultaneously determined. The median values of the microbial populations were obtained, and the data were analyzed per the sampling site by using the Wilcoxon and Kruskal-Wallis tests. The median of aerobic plate count content in tomatoes obtained from supermarkets ranged between 2.2 and 4.4 log CFU/g, and in markets from 2.9 to 4.8 log CFU/g. For molds and yeasts, the tomatoes from supermarkets (2.0 to 4.1 log CFU/g) and markets (1.5 to 4.5 log CFU/g) showed similar contents. Regardless of the sampling site, the values of total coliforms were very low, ranging from 1.0 to 1.8 log CFU/g. E. coli was detected in 5.4 and 20.1% of samples from supermarkets and markets, respectively; in both sites, the content was low (0.3 to 5.8 most probable number per g). The incidence of Salmonella was 14.1% in supermarkets and 7.8% in local markets. The results obtained from this investigation highlight the elevated risk for consumer health associated with the ingestion of cherry tomatoes.

Survey of Foodborne Pathogens, Aerobic Plate Counts, Total Coliform Counts, and Escherichia coli Counts in Leafy Greens, Sprouts, and Melons Marketed in the United States.

The objective of this research was to assess the microbiological status of leafy greens, sprouts, and melons from U.S. markets. A total of 14,183 samples of leafy greens, 2,652 samples of sprouts, and 3,411 samples of melons were collected throughout the United States from 2009 to 2014. The samples were analyzed for aerobic plate counts, total coliform counts, Escherichia coli counts, and the presence and levels of Salmonella, Shigella, Listeria monocytogenes, and Shiga toxin-producing E. coli (STEC), depending on the year and type of produce. Among the leafy greens, no E. coli O157:H7 or non-O157 STEC were detected from iceberg lettuce samples. The overall prevalences of Salmonella, E. coli O157:H7, non-O157 STEC, and L. monocytogenes in the 14,183 samples of leafy greens were 0.05, 0.01, 0.07, and 0.11%, respectively. Among sprout samples, no Salmonella or E. coli O157:H7 was detected, and the overall prevalences of non-O157 STEC and L. monocytogenes were 0.04 and 0.11%, respectively. Among melon samples, no Salmonella was detected from cucumbers, no L. monocytogenes was detected from cantaloupes, and the overall prevalences of Salmonella and L. monocytogenes were 0.12 and 0.23%, respectively. L. monocytogenes levels were 0.4 to 1,470 most probable number (MPN)/g in leafy greens, 0.36 to 1,100 MPN/g in sprouts, and <0.03 to 150 MPN/g in melons, and most positive samples had low levels of these pathogens. The isolates from these foods were very diverse genetically. Foodborne pathogens, including Salmonella, STEC, and L. monocytogenes, had relatively low prevalences in the produce surveyed. Because these foods are usually consumed raw, measures should be taken to significantly minimize the presence and levels of human pathogens.

Monitoring Shelf Life of Pasteurized Whole Milk Under Refrigerated Storage Conditions: Predictive Models for Quality Loss.

The shelf life of pasteurized milk is generally determined through microbiological analysis. The objective of this study was to correlate microbial quality parameters then to design predictive models for shelf life of pasteurized milk. We analyzed pasteurized milk (3.9% fat) for aerobic plate counts (APCs), psychrotrophic bacteria counts (PBCs), and Bacillus spp. counts at 5, 7, 10, 13, 15, and 19 (±1 °C) to the end of storage time. We also monitored titratable acidity, pH, and, lipase, and protease activity and correlated this with APC, which is the principal index defining shelf life. Results indicate that the shelf life of pasteurized milk was 24, 36, and 72 h at 19, 15, and 13 °C respectively, as determined by APC and acidity indicators. However, milk stored at lower temperatures of 5, 7, and 10 °C had longer shelf life of 30, 24, and 12 d, respectively. A sharp increase in titratable acidity, while decrease pH were observed when APCs reached 5.0 log10 CFU/mL at all storage temperatures. Lipase and protease activities increased with storage temperature. At 5 and 7 °C, however, protease activity was very low. Therefore, we eliminated this parameter from our quality parameters as a potential spoilage indicator. PRACTICAL APPLICATION: Findings of this research are useful for monitoring the quality of commercial pasteurized milk, particularly in locations where environmental conditions make longer storage difficult. The study also provides valuable information for development of colorimetric shelf life indicators.

Refrigeration of Oyster Shellstock: Conditions Which Minimize the Outgrowth of Vibrio vulnificus.

The multiplication of Vibrio vulnificus in summer harvest oyster shellstock held without refrigeration was followed over a 14 h postharvest period. Mean (n = 7) increases were 0,75, 1.30, 1.74, and 1.94 log units at 3.5 h, 7 h, 10.5 h, and 14 h postharvest, respectively. Aerobic plate counts (spread plates on plate count agar [PCA] containing 1% NaCl, 25°C) but not standard plate counts (pour plates, PCA, 35°C) showed a similar trend in increase. Reducing the time oyster shellstock remains outside refrigeration can decrease consumer exposure to high numbers of V. vulnificus , but shellstock must be cooled immediately after harvest to eliminate postharvest growth of this bacterium.