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Biltong is a South African air-dried beef product that does not have a heat lethality step, but rather relies on marinade chemistry (low pH from vinegar, ~2% salt, spices/pepper) in combination with drying at ambient temperature and low humidity to achieve microbial reduction during processing. Culture-dependent and culture-independent microbiome methodologies were used to determine the changes in the microbial community at each step during biltong processing through 8 days of drying. Culture-dependent analysis was conducted using agar-based methods to recover viable bacteria from each step in the biltong process that were identified with 16S rRNA PCR, sequencing, and BLAST searching of the NCBI nucleotide database. DNA was extracted from samples taken from the laboratory meat processing environment, biltong marinade, and beef samples at three stages of processing (post-marinade, day 4, and day 8). In all, 87 samples collected from two biltong trials with beef obtained from each of three separate meat processors (n = six trials) were amplified, sequenced with Illumina HiSeq, and evaluated with bioinformatic analysis for a culture-independent approach. Both culture-dependent and independent methodologies show a more diverse population of bacteria present on the vacuum-packaged chilled raw beef that reduces in diversity during biltong processing. The main genera present after processing were identified as Latilactobacillus sp., Lactococcus sp., and Carnobacterium sp. The high prevalence of these organisms is consistent with extended cold-storage of vacuum-packaged beef (from packers, to wholesalers, to end users), growth of psychrotrophs at refrigeration temperatures (Latilactobacillus sp., Carnobacterium sp.), and survival during biltong processing (Latilactobacillus sakei). The presence of these organisms on raw beef and their growth during conditions of beef storage appears to 'front-load' the raw beef with non-pathogenic organisms that are present at high levels leading into biltong processing. As shown in our prior study on the use of surrogate organisms, L. sakei is resistant to the biltong process (i.e., 2-log reduction), whereas Carnobacterium sp. demonstrated a 5-log reduction in the process; the recovery of either psychrotroph after biltong processing may be dependent on which was more prevalent on the raw beef. This phenomenon of psychrotrophic bloom during refrigerated storage of raw beef may result in a natural microbial suppression of mesophilic foodborne pathogens that are further reduced during biltong processing and contributes to the safety of this type of air-dried beef.
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Validation studies conducted within a food processing facility using surrogate organisms could better represent the manufacturing process than controlled laboratory studies with pathogenic bacteria on precision equipment in a BSL-2 lab. The objectives of this project were to examine potential surrogate bacteria during biltong processing, conduct biltong surrogate validation lethality studies, and measure critical factors and intrinsic parameters during processing. Beef pieces (1.9 cm × 5.1 cm × 7.6 cm) were inoculated with four-strain mixtures of Carnobacterium divergens/C. gallinarum, Pediococcus acidilactici/P. pentosaceous, and Biotype 1 E. coli ATCC BAA (-1427, -1428, -1429, and -1430), as well as a two-strain mixture of Latilactobacillus sakei and other commercially available individual bacterial cultures (P. acidilactici Saga200/Kerry Foods; Enterococcus faecium 201224-016/Vivolac Cultures). Inoculated beef was vacuum-tumbled in marinade and dried in a humidity-controlled oven for 8−10 days (24.9 °C; 55% relative humidity). Microbial enumeration of surviving surrogate bacteria and evaluation of intrinsic factors (water activity, pH, and salt concentration) were performed post inoculation, post marination, and after 2, 4, 6, 8, and 10 days of drying. Trials were performed in duplicate replication with triplicate samples per sampling time and analyzed by one-way RM-ANOVA. Trials conducted with E. faecium, Pediococcus spp., and L. sakei never demonstrated more than 2 log reduction during the biltong process. However, Carnobacterium achieved a >5 log (5.85 log) reduction over a drying period of 8 days and aligned with the reductions observed in previous trials with pathogenic bacteria (Salmonella, E. coli O157:H7, L. monocytogenes, and S. aureus) in biltong validation studies. Studies comparing resuspended freeze-dried or frozen cells vs. freshly grown cells for beef inoculation showed no significant differences during biltong processing. Carnobacterium spp. would be an effective nonpathogenic in-plant surrogate to monitor microbial safety that mimics the response of pathogenic bacteria to validate biltong processing within a manufacturer's own facility.
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Biltong is a dry beef product that is manufactured without a heat lethality step, raising concerns of whether effective microbial pathogen reduction can occur during biltong processing. Raw beef inoculated with 4-strain cocktails of either E. coli O157:H7, Listeria monocytogenes, or Staphylococcus aureus, and processed with a standard biltong process, were shown to incur a >5-log reduction in 6−8 days after marination by vacuum-tumbling for 30 min in vinegar, salt, spices (coriander, pepper) when dried at 23.9 °C (75 °F) at 55% relative humidity (RH). Pathogenic challenge strains were acid-adapted in media containing 1% glucose to ensure that the process was sufficiently robust to inhibit acid tolerant strains. Internal water activity (Aw) reached < 0.85 at 5-log reduction levels, ensuring that conditions were lower than that which would support bacterial growth, or toxin production by S. aureus should it be internalized during vacuum tumbling. This was further confirmed by ELISA testing for staphylococcal enterotoxins A and B (SEA, SEB) after marination and again after 10 days of drying whereby levels were lower than initial post-marination levels. Comparison of log reduction curves obtained for E. coli O157:H7, L. monocytogenes, S. aureus, and Salmonella (prior study) showed that microbial reduction was not significantly different (p < 0.05) demonstrating that even without a heat lethality step, the biltong process we examined produces a safe beef product according to USDA-FSIS guidelines.
RESUMEN
Salt (sodium chloride, NaCl) is commonly used in ready-to-eat (RTE) meat products such as biltong, a South African style dried beef product for flavor, enhanced moisture loss, and reduction of microbial growth. However, increased consumption of high sodium content foods is commonly associated with high blood pressure and heart disease. This study evaluated the use of alternative salts, potassium chloride (KCl) and calcium chloride (CaCl2) in the biltong marinade to achieve a ≥ 5-log reduction of Salmonella, a pathogen of concern in beef products. Beef pieces (1.9 cm × 5.1 cm × 7.6 cm) were inoculated with a five-serovar mixture of Salmonella (Salmonella Thompson 120, Salmonella Enteritidis H3527, Salmonella Typhimurium H3380, Salmonella Heidelberg F5038BG1, and Salmonella Hadar MF60404), vacuum-tumbled in a traditional biltong marinade of salt, spices, and vinegar containing either NaCl, KCl or CaCl2 (2.2% concentration) followed by an 8-10 day drying period at 23.9 °C (75 °F) and 55% relative humidity. Microbial enumeration of Salmonella was conducted following inoculation, after marination, and after 2, 4, 6, 8, and 10 days of drying in a humidity/temperature chamber. Biltong produced with CaCl2, NaCl, or KCl achieved a > 5-log reduction of Salmonella after 6, 7, and 8 days, respectively. The Salmonella reduction trends with biltong made with NaCl or CaCl2 were not significantly different (p < 0.05) while both were significantly different from that made with KCl (p > 0.05). Sodium, calcium, and potassium ion concentrations were measured using ion-specific electrode meters following biltong processing and drying. As expected, the biltong made with the corresponding salt had the most abundant ion in the sample. Regardless of the salt used in the marinade, the potassium ion levels were moderately elevated in all samples. This was determined to be from potassium levels naturally present in beef rather than from other ingredients. Sampling of several commercial brands of biltong for sodium content showed that some were significantly above the allowable level of claims made on package ingredient statements. The substitution of NaCl with KCl or CaCl2 during biltong processing can also provide a 5-log reduction of Salmonella to produce a safe product that can be marketed as a more healthy low-sodium food alternative that may appeal to consumers who need to reduce their blood pressure and are conscientious of sodium levels in their diet.
RESUMEN
In the US, dried beef products (beef jerky) are a popular snack product in which the manufacture often requires the use of a heat lethality step to provide adequate reduction of pathogens of concern (i.e., 5-log reduction of Salmonella as recommended by the United States Department of Agriculture Food Safety and Inspection Service (USDA-FSIS)). Biltong, a South African-style dried beef product, is manufactured with low heat and humidity. Our objectives were to examine processes for the manufacture of biltong that achieves a 5-log reduction of Salmonella without a heat lethality step and with, or without, the use of additional antimicrobials. Beef pieces (1.9 cm × 5.1 cm × 7.6 cm) were inoculated with a 5-serovar mixture of Salmonella (Salmonella Thompson 120, Salmonella Heidelberg F5038BG1, Salmonella Hadar MF60404, Salmonella Enteritidis H3527, and Salmonella Typhimurium H3380), dipped in antimicrobial solutions (lactic acid, acidified calcium sulfate, sodium acid sulfate) or water (no additional antimicrobial), and marinaded while vacuum tumbling and/or while held overnight at 5 °C. After marination, beef pieces were hung in an oven set at 22.2 °C (72 °F), 23.9 °C (75 °F), or 25 °C (77 °F) depending on the process, and maintained at 55% relative humidity. Beef samples were enumerated for Salmonella after inoculation, after dip treatment, after marination, and after 2, 4, 6, and 8 days of drying. Water activity was generally <0.85 by the end of 6-8 days of drying and weight loss was as high as 60%. Trials also examined salt concentration (1.7%, 2.2%, 2.7%) and marinade vinegar composition (2%, 3%, 4%) in the raw formulation. Nearly all approaches achieved 5-log10 reduction of Salmonella and was attributed to the manner of microbial enumeration eliminating the effects of microbial concentration on dried beef due to moisture loss. All trials were run as multiple replications and statistical analysis of treatments were determined by repeated measures analysis of variance (RM-ANOVA) to determine significant differences (p < 0.05). We believe this is the first published report of a biltong process achieving >5.0 log10 reduction of Salmonella which is a process validation requirement of USDA-FSIS for the sale of dried beef in the USA.
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INTRODUCTION: Shiga toxin-producing Escherichia coli (STEC) are foodborne pathogens that may cause diarrhoeal outbreaks and occasionally are associated with haemolytic-uraemic syndrome (HUS). We report on STEC O26:H11 associated with a cluster of four HUS cases in South Africa in 2017. METHODOLOGY: All case-patients were female and aged 5 years and under. Standard microbiological tests were performed for culture and identification of STEC from specimens (human stool and food samples). Further analysis of genomic DNA extracted from bacterial cultures and specimens included PCR for specific virulence genes, whole-genome sequencing and shotgun metagenomic sequencing. RESULTS: For 2/4 cases, stool specimens revealed STEC O26:H11 containing eae, stx2a and stx2b virulence genes. All food samples were found to be negative for STEC. No epidemiological links could be established between the HUS cases. Dried meat products were the leading food item suspected to be the vehicle of transmission for these cases, as 3/4 case-patients reported they had eaten this. However, testing of dried meat products could not confirm this. CONCLUSION: Since STEC infection does not always lead to severe symptoms, it is possible that many more cases were associated with this cluster and largely went unrecognized.
RESUMEN
Two infrared systems set at an intensity of 4777 W/m2 with peak emission wavelengths of 2.5 and 3.5 µm were used to produce biltong by drying differently pre-treated meat. In addition to meat texture and colour, the microbial quality of the biltong produced was assessed by quantifying viable heterotrophic microorganisms using a most probable number (MPN) method and by verifying the presence of presumptive Escherichia coli in samples produced using infrared and conventional convective drying. The two infrared drying systems reduced the heterotrophic microbial burden from 5.11 log10 MPN/g to 2.89 log10 MPN/g (2.5 µm) and 3.42 log10 MPN/g (3.5 µm), respectively. The infrared systems achieved an up to one log higher MPN/g reduction than the convective system. In biltong samples produced by short wavelength (2.5 µm) infrared drying, E. coli was not detectable. This study demonstrates that the use of short wavelength infrared drying is a potential alternative to conventional convective drying by improving the microbiological quality of biltong products while at the same time delivering products of satisfactory quality.
RESUMEN
The microbiological and physicochemical characteristics of several commercial beef, kudu and springbok biltong samples from South Africa were assessed in this study. Analysis of samples allowed their differentiation into 'dry' and 'moist' samples. Dry biltong showed low moisture content (21.5-25.3 g/100 g), a low water activity (0.65-0.68) and a high salt content (5.5-7.9 g/100 g), while moist biltong showed a higher moisture content (35.1-42.8 g/100 g), a higher water activity (0.85 to 0.89) and a lower salt content (3.8-5.6 g/100 g). The pH value did not vary significantly between both groups (5.00-6.26). The results showed that dry biltong presented a low total plate count (TPC) content, a high level of lactic acid bacteria (LAB) and a high level of D-lactic acid as compared to moist biltong. These results suggest that dry biltong complies with the standard hygienic quality (TPC<7 log cfu/g in agreement with the Food Standards Agency), while moist biltong samples generally showed a low content of D-lactic acid and a low ratio LAB/TPC.