Effects of a novel three-step sous-vide cooking and subsequent chilled storage on the microbiota of beef steaks.

This study aimed to evaluate a novel three-step sous-vide (SV) method on bacterial growth and diversity, and its relationship to product storage life. Vacuum-packed naturally contaminated steaks were sequentially cooked at 39 °C (1 h), 49 °C (1 h), and 59 °C (4 h), then stored at -1.5 and 2 °C for 28 d, with a single-step SV at 59 °C for 4 h for comparison. None of the seven indicator bacteria tested proliferated (P > .05) after incubation at 39 or 49 °C; microbial diversity was also unaffected. Bacterial load was reduced (P < .05) by up to 5 log units with both (P > .05) SV methods. The odour of all steaks remained acceptable on d 28. Unexpectedly, during storage, Pseudomonas, not lactic acid bacteria, dominated the microbiota on steaks cooked by either SV method, likely due to the temperature shift-induced lag phase and/or heat sensitivity of psychrotrophic bacteria. In conclusion, the three-step SV did not lead to bacterial proliferation or compromise the storage life of cooked products.

Growth of Carnobacterium spp. isolated from chilled vacuum-packaged meat under relevant acidic conditions.

Carnobacterium spp. are frequently isolated from vacuum-packaged (VP) meat. Specific strains of Carnobacterium and their growth characteristics may be associated with the storage life of such products. This study investigated the growth of 44 Carnobacterium isolates obtained from VP meat cuts produced at three Canadian abattoirs (A, B and C) under the following conditions: pH 5.4, 6.2 and 7.4; lactic acid at 60 and 90 mM; acetic acid at 33.6 mM. Whole genome sequencing was performed for all 44 isolates and a core genome phylogenetic tree was created to identify strain variability among isolates from different abattoirs. The isolates were clustered into 11 groups. All isolates from abattoirs B and C were identified as C. divergens, while the isolates from abattoir A included both C. maltaromaticum and C. divergens at equal proportions. C. divergens isolates from abattoir A belonged to two phylogenetic groups and none of them was found in the phylogenetic groups containing isolates from abattoirs B or C. Whole genome sequencing revealed that identical strains were isolated from different samples obtained at the same abattoir. The mean growth rate and maximum population density of the C. maltaromaticum isolates were lower than those of the C. divergens isolates. C. divergens isolates from abattoir A had higher growth rates and maximum population density than those from abattoirs B and C. In conclusion, growth characteristic and whole genome analysis both demonstrated strain variability of Carnobacterium among abattoirs, which could be a result of the difference in the antimicrobial interventions used for carcasses at different abattoirs, and may be associated with different storage lives of VP meats produced from different abattoirs.

Microbiological effects of a routine treatment for decontaminating hide-on carcasses at a large beef packing plant.

To investigate the microbiological effects of a hide-on carcass decontaminating treatment recently implemented at a beef packing plant, carcasses undergoing routine processing at the plant were sampled during successive periods in January/February, April/May, and September/October. During each period, samples were collected from carcasses before and after the decontamination of hide-on carcasses, after skinning, before decontamination of the skinned carcasses, and at the end of the carcass dressing process. At each stage of processing during each period, samples were obtained by swabbing an area of 1,000 cm(2) on each of 25 carcasses. Aerobes, coliforms, and Escherichia coli were enumerated. In most samples, coliforms were predominantly E. coli. In all three periods, the log mean numbers of aerobes and E. coli recovered from hides before decontamination were between 6.6 and 6.8 and between 5.3 and 5.9 log CFU/1,000 cm(2), respectively. The log mean numbers of aerobes recovered from decontaminated hides were 6.6 log CFU/1,000 cm(2) in January/February and April/May but 5.4 log CFU/1,000 cm(2) in September/October. The log total numbers of E. coli recovered from decontaminated hides in January/February and April/May were 2.4 and 3.8 log CFU/25,000 cm(2), respectively, but no E. coli was recovered from such carcasses in September/October. Log total numbers of aerobes and E. coli recovered from skinned or dressed carcasses were mostly >4 and between 1 and 2 log CFU/25,000 cm(2), respectively. Typing of 480 E. coli isolates by multiple-locus variable-number tandem repeat analysis (MLVA) identified 218 MLVA types. Most isolates recovered from carcasses in different periods or at different stages of processing were of different MLVA types. However, small numbers of MLVA types were recovered in more than one period or from both hides before and after decontamination and skinned or dressed carcasses. The findings show that the hide-decontaminating treatment disrupted the usual transfer of E. coli from hides to meat surfaces during carcass skinning.

Effects of meat pH on growth of 11 species of psychrotolerant clostridia on vacuum packaged beef and blown pack spoilage of the product.

The aim of the study was to determine the effects of meat pH on the abilities of 11 psychrotolerant Clostridium spp. to grow on, and to possibly cause blown pack spoilage of vacuum packaged beef. Beef steaks of pH 5.4-5.6, 5.7-5.9 or ≥6.0, i.e. of normal, intermediate or high pH were prepared and vacuum packaged. Groups of 3 steaks of the same pH range were inoculated with log phase cultures of Clostridium algoriphilum, Clostridium algidixylanolyticum, Clostridium bowmanii, Clostridium estertheticum, Clostridium frigoris, Clostridium frigidicarnis, Clostridium gasigenes, Clostridium lacusfryxellense, Clostridium psychrophilum, Clostridium tagluense or Clostridium vincentii. Each pack was resealed immediately after the steak was inoculated, and pack volumes were determined by water displacement, immediately after resealing and at intervals during storage at 2 °C for 56 days. All of the clostridia grew in packs of high pH beef but none caused pack swelling. Packs of intermediate pH beef inoculated with C. estertheticum began to swell after 14 days, with a mean rate of increase of pack volumes of 6.80 ml/day. One pack of intermediate pH beef inoculated with C. frigoris was swollen after 37 days. Packs of normal pH beef that had been inoculated with C. estertheticum began swelling after 14 days with a mean rate of increase of pack volumes of 7.70 ml/day. Packs of normal or intermediate pH beef inoculated with other clostridia did not swell. After storage, the numbers of most Clostridium spp., as determined by real-time PCR were greater on beef of high pH than of lower pH values, but the numbers of C. frigidicarnis and C. lacusfryxellense were highest on intermediate pH meat, the numbers of C. estertheticum were higher on meat of lower than of high pH, and the numbers of C. tagluense were the same on meat of all pH values. With high pH meat, glucose was reduced to very low level in rinse fluids from packs that had been inoculated with any Clostridium sp. With intermediate and normal pH meat, glucose was reduced to very low concentrations in only rinse fluids from beef that had been inoculated with C. estertheticum.

Substrate utilization during incubation in meat juice medium of psychrotolerant clostridia associated with blown pack spoilage.

Several new species of psychrophilic or psychrotolerant clostridia have been identified in recent years. Some of these may be involved in 'blown pack' spoilage (BPS) of vacuum packaged beef. Organisms that cause BPS must produce large volumes of gas while utilizing substrates available in raw meat. Therefore, Clostridium algoriphilum, Clostridium algidixylanolyticum, Clostridium bowmanii, Clostridium frigoris, Clostridium frigidicarnis, Clostridium gasigenes, Clostridium lacusfryxellense, Clostridium psychrophilum, Clostridium tagluense and Clostridium vincentii were grown in meat juice medium (MJM), and changes in substrate concentrations were monitored to assess the potential for gas production by each organism. All 10 species were able to grow exponentially on glucose with simultaneous hydrolysis of glycogen, reaching maximum values for absorbance at 600 nm of 0.3-1.90. All ceased growing when glucose and glycogen were still detectable in the growth medium. C. frigidicarnis utilized most of the amino acids available in MJM and reduced the concentration of total amino acids by 10 mM. The other 9 species caused little or no reduction in amino acid concentrations. C. algidixylanolyticum and C. frigidicarnis utilized glucose, glycogen and lactate simultaneously during growth and after growth ceased. C. algoriphilum and C. frigoris commenced utilization of lactate, while continuing utilization of glucose and glycogen, only after growth ceased, but utilization of lactate by C. algoriphilum was weak. C. psychrophilum ceased utilization of glucose and glycogen but initiated weak utilization of lactate after growth ceased. The other 5 species did not utilize any substrate after growth ceased. The utilization of glucose, glycogen and relatively large amounts of lactate by C. algidixylanolyticum, C. frigoris, and C. frigidicarnis after growth ceased indicates that these organisms have the potential to cause BPS. The other 7 species appear to lack such potential.

Use of propidium monoazide and quantitative PCR for differentiation of viable Escherichia coli from E. coli killed by mild or pasteurizing heat treatments.

Suspensions of Escherichia coli in peptone water were heated at temperatures between 52 and 90 °C, inclusive. Samples withdrawn at suitable times were not or were treated with propidium monoazide (PMA) or deoxycholate then PMA before extraction of DNA. DNA was quantified by real-time PCR for estimation of the numbers of E. coli from which template DNA for the PCR was obtained. Numbers of viable E. coli in suspensions at the times of sampling were determined from plate counts. For samples from suspensions heated at temperatures ≥ 52 ≤ 72 °C, PCR cycle threshold (Ct) values were little or no different for DNA from corresponding samples that were or were not treated with PMA. PMA treatment of samples heated to ≥ 80 °C largely inactivated E. coli DNA for PCR. When samples heated to ≤ 72 °C were treated with deoxycholate before treatment with PMA, Ct values for treated samples were greater than the Ct values for the corresponding untreated samples. Similar results were obtained with E. coli suspended in milk or fluid from ground beef pummeled with diluent. The results indicate that cells killed by heating to ≥ 80 °C are permeable to PMA, but most cells killed by heating to ≤ 72 °C are not. However, treatment with deoxycholate renders a substantial fraction of the latter cells permeable to PMA. Numbers of viable or dead E. coli can then be estimated from Ct values for samples not treated or treated with deoxycholate and PMA, provided viable cells are ≥ 1% of the total.