The efficiency of Lactiplantibacillus plantarum S61 strain as protective cultures in ground beef against foodborne pathogen Escherichia coli.

The aim of the study was the bio-control effectiveness of the Lactiplantibacillus plantarum S61 strain, isolated from traditional fermenting green olives, against Escherichia coli B805 in ground beef. The bio-control effect of L. plantarum S61 against E. coli B805 was evaluated in ground meat during storage under refrigeration at 4 °C. The results showed that L. plantarum S61 reduced the biomass of pathogenic bacteria (E. coli) in ground meat during 10 days of storage at 4 °C. Moreover, the treatment with L. plantarum S61 has no adverse effect on the sensory properties of ground meat after 10 days of storage at 4 °C. The treatment with L. plantarum S61 and storage at 4 °C effectively decreases the growth and risk of pathogenic bacteria in ground meat and, consequently, increases the product's shelf life. Therefore, the application of L. plantarum S61 during the storage of ground meat beef may help reduce the use of chemical preservatives in meat products. Consequently, L. plantarum S61 can be applied as a bio-control agent against spoilage and pathogenic bacteria in meat and meat products.

A predictive growth model for Clostridium botulinum during cooling of cooked uncured ground beef.

A dynamic model to predict the germination and outgrowth of Clostridium botulinum spores in cooked ground beef was presented. Raw ground beef was inoculated with a ten-strain C. botulinum spore cocktail to achieve approximately 2 log spores/g. The inoculated ground beef was vacuum packaged, cooked to 71 °C to heat shock the spores, cooled to below 10 °C, and incubated isothermally at temperatures from 10 to 46 °C. C. botulinum growth was quantified and fitted into the primary Baranyi Model. Secondary models were fitted to maximum specific growth rate and lag phase duration using Modified Ratkowsky equation (R2 0.96) and hyperbolic function (R2 0.94), respectively. Similar experiments were also performed under non-isothermal (cooling) conditions. Acceptable zone prediction (APZ) analysis was conducted on growth data collected over 3 linear cooling regimes from the current study. The model performance (prediction errors) for all 22 validation data points collected in the current work were within the APZ limits (-1.0 to +0.5 log CFU/g). Additionally, two other growth data sets of C. botulinum reported in the literature were also subjected to the APZ analysis. In these validations, 20/22 and 10/14 predictions fell within the APZ limits. The model presented in this work can be employed to predict C. botulinum spore germination and growth in cooked uncured beef under non-isothermal conditions. The beef industry processors and food service organizations can utilize this predictive microbial model for cooling deviations and temperature abused situations and in developing customized process schedules for cooked, uncured beef products.

Rapid estimation of Salmonella enterica contamination level in ground beef - Application of the time-to-positivity method using a combination of molecular detection and direct plating.

Little progress has been made in decreasing the incidence rate of salmonellosis in the US over the past decade. Mitigating the contribution of contaminated raw meat to the salmonellosis incidence rate requires rapid methods for quantifying Salmonella, so that highly contaminated products can be removed before entering the food chain. Here we evaluated the use of Time-to-Positivity (TTP) as a rapid, semi-quantitative approach for estimating Salmonella contamination levels in ground beef. Growth rates of 14 Salmonella strains (inoculated at log 1 to -2 CFU/g) were characterized in lean ground beef mTSB enrichments and time-to-detection was determined using culture and molecular detection methods. Enrichments were sampled at five timepoints and results were used to construct a prediction model of estimated contamination level by TTP (superscript indicates time in hours) defined as TTP4: ≥5 CFU/g; TTP6: ≤5, ≥1 CFU/g; TTP8: ≤1, ≥0.01 CFU/g; with samples negative at 8 h estimated ≤0.01 CFU/g. Model performance measures showed high sensitivity (100%) and specificity (83% and 93% for two detection methods) for samples with a TTP4, with false negative rates of 0%.

Effects of lyophilized black carrot (Daucus carota L.) water extract on the shelf life, physico-chemical and microbiological quality of high-oxygen modified atmosphere packaged (HiOx-MAP) ground beef.

In the present study, firstly, various properties of lyophilized water extracts (LBCWE) produced from fresh black carrot were determined. LBCWE was observed to be a rich source of monomeric anthocyanins (1188.40 ± 17.38 mg C3G/100 g; n = 4) and phenolics (2733.83 ± 17.78 mg GAE/100 g, n = 4). Secondly, ground beef containing LBCWE (Control, 100, 200 and 300 ppm) and packaged in HiOx-MAP (80% O2 + 20% CO2) was evaluated in terms of lipid oxidation, metmyoglobin (MetMb), color, pH and microbial counts during storage at 2.0 ± 0.5 °C for 12 days. By increasing level of LBCWE, the pH, lipid oxidation, MetMb and microbial counts were decreased (P < 0.01). The LBCWE significantly affected the color and microbial count parameters (P < 0.01). The highest redness and lowest microbial growth during storage was in the 300 ppm LBCWE group (P < 0.05). On the 12th day of storage, Pseudomonas and Enterobactericeae decreased 1.24 log and 1.46 log units in this group according to control. The shelf life of ground beef can be extended by 3 days with MAP + 300 ppm extract application. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13197-021-05044-1.

Isolation and characterization of non-O157 Shiga toxin-producing Escherichia coli (STEC) isolated from retail ground beef in Santiago, Chile.

Shiga toxin-producing Escherichia coli (STEC) is one of the main cause of foodborne disease worldwide, but isolation rates or characteristics of this bacteria from ground beef in Chile are unknown. The present study aimed to isolate and characterize non-O157 STEC from ground beef sold at retail in the city of Santiago, Chile. We analyzed 430 ground beef samples for the presence of STEC, and isolated the microorganism in 10% of samples (43/430). We obtained 56 isolates from the 43 positive samples; 55 of these (98.2%) fermented sorbitol. Most isolates (98.2%; 55/56) showed ß-glucoronidase activity, and only six (10.7%; 6/56) were resistant to tellurite. Among the virulence factors studied (stx1, stx2, eae, and hlyA), stx2 was the only virulence factor in 41% of the isolates (23/56), whereas 10.7% (6/56) of isolates carried a combination of three virulence factors (stx1 + stx2 + hlyA). None of the isolates carried the gene eae. Finally, isolates were neither serogroups O157 nor "big six". In conclusion, ground beef sold in Santiago, Chile is contaminated with STEC; however, further studies are required for understanding their virulence potential.

Strand-specific transcriptomes of Enterohemorrhagic Escherichia coli in response to interactions with ground beef microbiota: interactions between microorganisms in raw meat.

BACKGROUND: Enterohemorrhagic Escherichia coli (EHEC) are zoonotic agents associated with outbreaks worldwide. Growth of EHEC strains in ground beef could be inhibited by background microbiota that is present initially at levels greater than that of the pathogen E. coli. However, how the microbiota outcompetes the pathogenic bacteria is unknown. Our objective was to identify metabolic pathways of EHEC that were altered by natural microbiota in order to improve our understanding of the mechanisms controlling the growth and survival of EHECs in ground beef. RESULTS: Based on 16S metagenomics analysis, we identified the microbial community structure in our beef samples which was an essential preliminary for subtractively analyzing the gene expression of the EHEC strains. Then, we applied strand-specific RNA-seq to investigate the effects of this microbiota on the global gene expression of EHEC O2621765 and O157EDL933 strains by comparison with their behavior in beef meat without microbiota. In strain O2621765, the expression of genes connected with nitrate metabolism and nitrite detoxification, DNA repair, iron and nickel acquisition and carbohydrate metabolism, and numerous genes involved in amino acid metabolism were down-regulated. Further, the observed repression of ftsL and murF, involved respectively in building the cytokinetic ring apparatus and in synthesizing the cytoplasmic precursor of cell wall peptidoglycan, might help to explain the microbiota's inhibitory effect on EHECs. For strain O157EDL933, the induced expression of the genes implicated in detoxification and the general stress response and the repressed expression of the peR gene, a gene negatively associated with the virulence phenotype, might be linked to the survival and virulence of O157:H7 in ground beef with microbiota. CONCLUSION: In the present study, we show how RNA-Seq coupled with a 16S metagenomics analysis can be used to identify the effects of a complex microbial community on relevant functions of an individual microbe within it. These findings add to our understanding of the behavior of EHECs in ground beef. By measuring transcriptional responses of EHEC, we could identify putative targets which may be useful to develop new strategies to limit their shedding in ground meat thus reducing the risk of human illnesses.

Differences in inactivation of Escherichia coli O157:H7 strains in ground beef following repeated high pressure processing treatments and cold storage.

High pressure processing (HPP) is a safe non-thermal processing method to effectively improve food safety. In this study, HPP treatment followed by cold storage was investigated to reduce Escherichia coli O157:H7 in ground beef. Experiments were conducted using ground beef contaminated with six E. coli O157:H7 strains one at a time or as a cocktail. Control and inoculated ground beef samples were HPP at 25 °C, 35 °C, and 45 °C, at 400 MPa and pre-determined number of pressure cycles totaling a holding time of 15 min. Optimum HPP parameters were 25 °C, 400 MPa at five pressure cycles of 3 min each which achieved a 5-log reduction of E. coli O157:H7 in ground beef. Storing HPP processed ground beef at 4 °C or -20 °C further decreased (P < 0.05) the E. coli O157:H7 population. An effective HPP treatment (5-log reduction) was developed that could be used post-processing to reduce the risk associated with E. coli O157:H7 contamination in ground beef.

Inactivation of Shiga toxin-producing Escherichia coli in lean ground beef by gamma irradiation.

In this study the radiation resistance of 40 Shiga Toxin-Producing Escherichia coli (STEC) isolates which contained various combinations of the shiga toxin 1 (stx1), shiga toxin 2 (stx2), intimin (eae), and hemolysin (ehx) genes were determined. The STEC were suspended in lean ground beef and irradiated at 4 °C. D10 values, the radiation dose needed to reduce 1 log (90%) of a microorganism, ranged from 0.16 to 0.48 kGy, with a mean of 0.31 kGy for the 40 isolates. Isolates associated with illness outbreaks had a mean D10 of 0.27 kGy, while non-outbreak isolates had a mean D10 of 0.36 kGy (p < 0.05). The presence or absence of stx1, stx2, or both stx1 and 2 had no affect on D10 (p > 0.05). The presence (0.30 kGy) or absence (0.35 kGy) of ehx had no affect on D10 (p > 0.05). However, the mean D10 of isolates lacking eae (0.37 kGy) were significantly higher than those containing eae (0.27 kGy) (p < 0.05). There was no difference in D10 for isolates lacking eae regardless of whether or not they were associated with a foodborne illness outbreak (p > 0.05). It may be possible to use some of the STEC isolates which lacked eae, ehx, or both (D10 > 0.30) as avirulent surrogates in food irradiation research. The data presented in this study provides risk assessors data for metagenomic analysis as well as food and radiation processors with valuable information to control of STEC in meat.

Inactivation of a diverse set of shiga toxin-producing Escherichia coli in ground beef by high pressure processing.

Shiga toxin-producing Escherichia coli (STEC) are regularly implicated in foodborne illness outbreaks and recalls of ground beef. In this study we determined the High Pressure Processing (HPP) D10 value (the processing conditions needed to reduce the microbial population by 1 log) of 39 STEC isolates, including the "big six" serovars, O104 and O157:H7. STEC isolates included those isolated from animals and environmental sources in addition to those associated with illness in humans. Individual STEC were inoculated into 80% lean ground beef and treated with HPP (350 MPa, 4 °C, up to 40 min). The mean D10 was 9.74 min, with a range of 0.89-25.70 min. The D10 of the STEC involved in human illness was 9.25 vs. 10.40 min for those not involved in human illness (p > 0.05). The presence or absence of genes encoding virulence factors (e.g. Shiga toxin 1 or 2, intimin, or enterohemolysin) had no effect on the HPP D10 (p > 0.05). The high D10 of some STEC involved in human illness should be considered in selecting HPP processing parameters for ground beef. This study demonstrates the heterogeneity of STEC resistance to HPP. Risk assessors and the food industry can use this information to provide safer meat products to consumers.

Effect of high pressure processing on the survival of Shiga toxin-producing Escherichia coli (Big Six vs. O157:H7) in ground beef.

High pressure processing (HPP) is a safe and effective technology for improving food safety. Non-O157:H7 Shiga Toxin-producing Escherichia coli (STEC) have been increasingly implicated in foodborne illness outbreaks and recalls, and the USDA Food Safety Inspection Service (FSIS) has designated them as adulterants in meat (e.g. ground beef). In this study we compared the inactivation of multi-isolate cocktails of E. coli O157:H7 versus the non-O157:H7 STEC "Big Six" (i.e. O26, O45, O103, O111, O121, and O145) in ground beef (83% lean) using HPP at refrigeration temperature (4-7 °C). A >5-log CFU/g inactivation of both the Big Six and O157:H7 cocktails were observed at 450 MPa for 15 min. In general, the Big Six cocktail was found more sensitive to pressure stress (p < 0.05). In contrast, HPP treatment at 250 MPa (30 min) inactivated only 2.3 log of the Big Six versus 1.0 log of O157:H7. HPP treatment at 350 MPa (30 min) inactivated 4.7 log of the Big Six vs. 3.2 log of O157:H7. Multiple-cycle HPP cycles (250 or 350 MPa, three 5 min treatments) did not result in a 5 log reduction of the non-O157:H7 or O157:H7 STEC. Our results indicate that HPP inactivation parameters which are effective for O157:H7 STEC can be used for the non-O157:H7 Big Six isolates in ground beef.

Use of ß-cyclodextrin and activated carbon for quantification of Salmonella enterica ser. Enteritidis from ground beef by conventional PCR without enrichment.

The high level of PCR inhibitors present in ground beef is a major factor that affects molecular based techniques, such as the polymerase chain reaction (PCR), for the detection of Salmonella enterica. In this study, the use of ß-cyclodextrin and milk protein coated activated carbon (MP-CAC) allowed the PCR to detect low numbers of Salmonella seeded into ground beef without enrichment of samples. invA was used as target gene in the conventional PCR protocol. With 25 g samples of ground beef containing 7.0, 15, and 27% fat, treatment of stomached samples with 5.0, 10, and 15% ß-cyclodextrin respectively followed by treatment with MP-CAC, resulted in the detection of 3 CFU/g (equivalent to 75 CFU in a 25 g sample). The total assay time was 4.5 h. The methodology described in this study for the detection of S. enterica in ground beef without enrichment is rapid, sensitive, and has the potential to be applied to a number of complex food matrices to detect low numbers of food-borne bacterial pathogens.

Effect of Ground Beef Irradiation on Annual Nontyphoidal Salmonella and Escherichia coli O157 Burden and Direct Healthcare Costs in the United States: A Simulation Study.

Over 20% of E. coli O157 illnesses and over 5% of Salmonella illnesses are estimated to be attributable to beef consumption in the United States. Irradiating ground beef is one possible method to reduce disease burden. We simulated the effect of ground beef irradiation on illnesses, hospitalizations, deaths, and direct healthcare costs from ground beef-associated E. coli O157 and Salmonella illnesses in the United States. To estimate the fraction of illnesses, hospitalizations, deaths, and direct healthcare costs preventable by ground beef irradiation, we multiplied the disease burden attributable to ground beef; the estimated percentage of ground beef sold that is not currently irradiated; the percentage of unirradiated ground beef that would be irradiated; and the percentage reduction in risk of illness after irradiation. We multiplied this fraction by estimates of burden and direct healthcare costs to calculate the numbers or amounts averted. Model inputs were obtained from the literature and expert opinion. We used Monte Carlo simulation to incorporate uncertainty in inputs into model estimates. Simulation outcomes were summarized with means and 95% uncertainty intervals (UI). Irradiating 50% of the currently unirradiated ground beef supply would avert 3,285 (95% UI: 624-9,977) E. coli O157 illnesses, 135 (95% UI: 24-397) hospitalizations, 197 (95% UI: 34-631) hemolytic uremic syndrome cases, 2 (95% UI: 0-16) deaths, and $2,972,656 (95% UI: $254,708-$14,496,916) in direct healthcare costs annually. For Salmonella, irradiation would avert 20,308 (95% UI: 9,858-38,903) illnesses, 400 (95% UI: 158-834) hospitalizations, 6 (95% UI: 0-18) deaths, and $7,318,632 (95% UI: $1,436,141-$26,439,493) in direct healthcare costs. Increasing ground beef irradiation could reduce E. coli O157 and Salmonella burden in the United States. Additional studies should assess whether targeted irradiation of higher-risk ground beef products could prevent similar numbers of illnesses with less total product irradiated.

In vitro antimicrobial activity of ginseng extract against Staphylococcus aureus, Salmonella Typhimurium and Listeria monocytogenes and its inhibitory effects on these pathogens in cooked ground beef.

Present study is focused on exploring in vitro antimicrobial activity of ginseng extract (GE) against Staphylococcus aureus, Salmonella Typhimurium and Listeria monocytogenes and their biofilm forming ability, then determining its antimicrobial efficiency during the storage (4 and 10 °C) of cooked ground beef contaminated with these pathogens. Results revealed that the minimum inhibitory concentration (MIC) for S. aureus, S. typhimurium and L. monocytogenes were 90, 70 and 40 mg/mL, respectively. GE was also able to inhibit biofilm production by S. aureus and L. monocytogenes, whereas S. typhimurium did not produce biofilm with or without GE application. On the other hand, the results of GE aplication in ground beef indicated that GE incorporation at 0.5% or more reduced S. aureus and S. Typhmurium counts in cooked ground beef at the end of 30 d storage at 4 °C (P < 0.05). Using 1% or more GE totally inhibited S. Typhmurium after 15 d storage at 4 °C (P < 0.05). However, GE application did not affect S. aureus or S. Typhmurium counts in cooked ground beef stored at 10 °C. Furthermore, incorporation of 0.5% or more GE generally inhibited L. monocytogenes growth in cooked ground beef at the end of storage at both 4 and 10 °C (P < 0.05). In general, pH was lower in samples with GE than those without GE regardless of differences in the type of tested pathogen, storage time and temperatures (P < 0.05). Although similar aw was generally obtained on processing day, lower aw was observed in GE containing groups at the end of storage for both storage temperatures (P < 0.05). Oxidation reduction potential (ORP) in all groups generally increased during storage at both storage temperatures (P < 0.05). In conclusion, our results demonstrated that GE has an important role in controlling growth of tested pathogens and may be used as a natural agent by the meat industry to inhibit the growth of food-borne pathogens in cooked processed meat products during storage.

Modelling and numerical methods for identifying low-level adulteration in ground beef using near-infrared hyperspectral imaging (NIR-HSI).

Owing to the inherent characteristics of ground beef, adulteration presents a substantial risk for suppliers and consumers alike. This study developed a robust and novel method for identifying replacement fraud in ground beef with beef liver, beef heart, and pork using Near Infrared-Hyperspectral Imaging (NIR-HSI) coupled with chemometric and other statistical methods. More specifically, NIR-HSI provided an efficient and accurate means of identifying each type of adulteration using the classification model Genetic Algorithm (GA) - Backpropagation Artificial Neural Network (BPANN), showing perfect sensitivity and specificity (a value of 1.00) for the calibration and the validation sets for all types of adulteration. As an alternative to chemometric analysis, Hyperspectral Imaging-Root Mean Square (HSI-RMS) value, based on the RMScut-off calculation, was determined to discriminate types of adulterations without the need of resource-intensive modelling. This HSI-RMS approach provides a simple-to-use method that avoids the complexity of HSI data processing and aims to directly understand the similarity between different spectra of one sample in the pixel level. Different types of adulteration show noticeable differences reflected in the HSI-RMS value (varying from 55 to 1439), which demonstrate the potential of HSI-RMS concept as a novel and valuable alternative for assessing the HSI data and facilitating the identification of adulterants.

Comparing Individual and Community-level Characteristics of People with Ground Beef-associated Salmonellosis and Other Ground Beef Eaters: A Case-control Analysis.

Salmonella is estimated to be the leading bacterial cause of U.S. domestically acquired foodborne illness. Large outbreaks of Salmonella attributed to ground beef have been reported in recent years. The demographic and sociodemographic characteristics of infected individuals linked to these outbreaks are poorly understood. We employed a retrospective case-control design; case-patients were people with laboratory-confirmed Salmonella infections linked to ground beef-associated outbreaks between 2012 and 2019, and controls were respondents to the 2018-2019 FoodNet Population Survey who reported eating ground beef and denied recent gastrointestinal illness. We used county-level CDC/ATSDR Social Vulnerability Index (SVI) to compare case-patient and controls. Case-patient status was regressed on county-level social vulnerability and individual-level demographic characteristics. We identified 376 case-patients and 1,321 controls in the FoodNet sites. Being a case-patient was associated with increased overall county-level social vulnerability (OR: 1.21 [95% CI: 1.07-1.36]) and socioeconomic vulnerability (OR: 1.24 [1.05-1.47]) when adjusted for individual-level demographics. Case-patient status was not strongly associated with the other SVI themes of household composition and disability, minority status and language, and housing type and transportation. Data on individual-level factors such as income, poverty, unemployment, and education could facilitate further analyses to understand this relationship.

Beef Consumption Is Associated with Higher Intakes and Adequacy of Key Nutrients in Older Adults Age 60+ Years: National Health and Nutrition Examination Survey 2011-2018 Analysis.

Beef is an important source of high-quality protein and several micronutrients, including iron, zinc, and B vitamins. We determined beef intake and its relationship with intakes of nutrients and their adequacy using 24 h dietary recall data from 5868 older adults. Usual intakes from foods were determined using the National Cancer Institute method, and the percent of the population below the estimated average requirement or above adequate intake was estimated. A high percentage of older adults did not meet nutrient recommendations for vitamin D (96%), choline (96%), vitamin E (84%), potassium (70%), calcium (63%), magnesium (60%), vitamin C (46%), vitamin A (39%), zinc (21%), vitamin B6 (19%), and folate (15%). About 68% of older adults were beef consumers with a mean intake of 56 g/day. Beef consumers had higher (p < 0.05) intakes of energy, protein, calcium, iron, phosphorus, selenium, sodium, zinc, thiamin, riboflavin, niacin, vitamin B12, and choline, and a higher (p < 0.05) proportion met nutrient recommendations for protein, calcium, copper, zinc, thiamin, folate, and vitamin B12 than non-consumers. Consumers of fresh, ground, and processed beef also had generally higher intakes and lower inadequacies of many nutrients depending on the beef type. In conclusion, older adults generally had poor nutrient adequacy from their diets, while beef consumers had higher nutrient intakes and adequacy for certain key nutrients, which are inherently generally available from beef or from foods consumed with beef.

Virulent shiga toxin-producing Escherichia coli (STEC) O157:H7 ST11 isolated from ground beef in Brazil.

In this study, a total of 248 ground beef samples were analyzed for the presence of Shiga toxin-producing Escherichia coli (STEC). Out of these samples, only one (0.4%) tested positive for STEC. Further analysis using PCR confirmed the presence of all tested genes associated with STEC, including stx1, stx2, eae, ehx, uid, rfbO157, and fliCH7 in this isolate. Interestingly, no STEC strains were detected in the remaining 100 beef cut samples or the 100 chicken cut samples, indicating the absence of detectable STEC contamination in those specific samples. The isolated strain exhibited significant cytotoxic activity in Vero cells, indicating its ability to produce cytotoxic Shiga toxins. To further investigate the strain, whole-genome sequencing (WGS) analyses were performed. The resistome analysis revealed the absence of acquired antimicrobial resistance genes, indicating a pan-susceptible phenotype. However, this strain presented chromosomal mutations in gyrA, gyrB, parC, parE, pmrA, pmrB, and folP. Plasmid analysis identified the presence of two plasmids, namely IncFIB(AP001918) and IncFII. The multi-locus sequence typing (MLST) identified the strain as belonging to sequence type (ST) 11, which is associated with E. coli O157:H7 strains. The virulome analysis confirmed the presence of several canonical virulence markers, including stx1, stx2, eae-g01-gamma, ehxA, stx1a-O157, and stx2a-O157. Overall, this study identified for the first time a rare occurrence of STEC contamination in ground beef, with the isolated strain belonging to the highly virulent O157:H7 serotype. These findings contribute to our understanding of STEC prevalence and characteristics in food samples, highlighting the importance of effective food safety measures to prevent potential health risks associated with STEC contamination.

Comparison of Gas Treatments of High Oxygen, Carbon Monoxide, and Nitric Oxide on Ground Beef Color in Modified Atmosphere Packaging.

The objective of this study was to evaluate the viability and performance of nitric oxide modified-atmosphere packaging (MAP) as a novel alternative to high oxygen and carbon monoxide MAP for ground beef. Packages of ground beef under high oxygen (HI-OX), carbon monoxide (CO), and nitric oxide (NO) atmospheres were evaluated for descriptive and instrumental color every 12 h during a 120 h display period. Surface myoglobin percentages, internal cooked color, thiobarbituric acid reactive substances (TBARS), and residual nitrite and nitrate were also evaluated. There were gas × time interactions for descriptive color, discoloration, a* values, b* values, deoxymyoglobin percentages, and metmyoglobin percentages (p < 0.05). There were also gas-type main effects for cooked color and TBARS (p < 0.05). Carbon monoxide maintained the most redness and least discoloration throughout the display period, while HI-OX started with a bright red color but rapidly browned (p < 0.05). Nitric oxide started as dark red to tannish-red but transitioned to a dull red (p < 0.05). However, NO had increased redness and a* values for internal cooked color (p < 0.05). Although CO outperformed NO packages, NO exhibited a unique color cycle warranting further research to optimize its use.

Contribution of Beef to Key Nutrient Intakes and Nutrient Adequacy in Pregnant and Lactating Women: NHANES 2011-2018 Analysis.

Beef is an important source of high-quality protein and several micronutrients, including iron, zinc, and B-vitamins. The objective was to assess the association of beef intake with nutrient intake and adequacy among pregnant and lactating women using 24-h dietary recall data. Usual intakes from foods were determined with the National Cancer Institute (NCI) method and % population below Estimated Average Requirement (EAR) or above Adequate Intake (AI) were estimated. A high proportion of pregnant and lactating women had inadequate intakes for vitamin D (94%), vitamin E (82%), vitamin C (52%), and vitamin A (50%), magnesium (35%), folate (31%), zinc (25%), and vitamin B6 (22%); only 4% and 35% met AI for choline and potassium, respectively. About 67% of pregnant and lactating women were beef consumers, consuming 49 g beef/day. Beef consumers had higher intakes (p < 0.05) of energy, protein, calcium, iron, phosphorus, selenium, sodium, zinc, thiamin, riboflavin, and niacin, and a higher proportion (p < 0.05) met nutrient recommendations for protein, calcium, iron, zinc, thiamin, riboflavin, niacin, vitamin B6, and vitamin B12 compared to non-consumers. In conclusion, pregnant and lactating women generally have inadequate nutrient intakes from their diets. Beef consumers have higher intakes and adequacy for certain nutrients, many of which are inherently available in beef or in foods eaten with beef.

Real-time PCR primers and probes for the detection of Shiga toxin genes, including novel subtypes.

Shiga toxin-producing Escherichia coli (STEC) are foodborne enteric pathogens. STEC are differentiated from other E. coli by detection of Shiga toxin (Stx) or its gene (stx). The established nomenclature of Stx identifies ten subtypes (Stx1a, Stx1c, Stxd, Stx2a to Stx2g). An additional nine subtypes have been reported and described (Stx1e, Stx2h to Stx2o). Many PCR protocols only detect a subset of Stx subtypes which limits their inclusivity. Here we describe a real-time PCR assay inclusive of the DNA sequences of representatives of all currently described Stx subtypes. A multiplex real-time PCR assay for detection of stx was developed using nine primers and four probes. Since the identification of STEC does not require differentiation of stx subtypes, the probes use the same fluorescent reporter to enable detection of multiple possible targets in a single reaction. The PCR mixture includes an internal positive control to detect inhibition of the reaction. Thus, the protocol can be performed on a two-channel real-time PCR platform. To reduce the biosafety risk inherent in the use of STEC cultures as process controls, the protocol also includes the option of a non-pathogenic E. coli transformant carrying a plasmid encoding the targeted fragment of the stx2a sequence. The inclusivity of the PCR was assessed against colonies of 137 STEC strains and one strain of Shigella dysenteriae, including strains carrying single copies of stx representing fourteen subtypes (stx1 a, c, d; stx2 a-j and o). Five additional subtypes (stx1e, 2k, 2l, 2m and 2n) were represented by E. coli transformed with plasmids encoding toxoid (enzymatically inactive A subunit) sequences. The exclusivity panel consisted of 70 bacteria, including 21 stx-negative E. coli. Suitability for food analysis was assessed with artificially inoculated ground beef, spinach, cheese, and apple cider. The real-time PCR generated positive results for all 19 stx subtypes, represented by colonies of STEC, S. dysenteriae and E. coli transformants carrying stx toxoid plasmids. Tests of exclusivity panel colonies were all negative. The real-time PCR detected the presence of stx in all inoculated food enrichments tested, and the presence of STEC was confirmed by isolation.