Javier Calzada

Effect of high-pressure treatment on the survival of Listeria monocytogenes Scott A in sliced vacuum-packaged Iberian and Serrano cured hams.

High-pressure treatment is useful for increasing the microbiological safety of ready-to-eat foods. With dry-cured hams, this treatment can be applied to the finished product after slicing and vacuum packaging. The effect of high-pressure treatment on the survival of inoculated Listeria monocytogenes Scott A and on the sensory characteristics of two Spanish dry-cured hams, Iberian and Serrano, was investigated. Ham slices were inoculated with L. monocytogenes at 6 x 10(6) CFU/g and held at 4 degrees C for 20 h before high-pressure treatment. During this holding period, the population of the pathogen declined by 0.44 and 0.51 log CFU/g in Iberian and Serrano hams, respectively. Treatment at 450 MPa for 10 min at 12 degrees C reduced L. monocytogenes populations by 1.50 and 1.16 log CFU/g in Iberian and Serrano hams, respectively. During the first week of storage at 4 or 8 degrees C, L. monocytogenes populations declined by an average 0.89 log CFU/g in pressurized Iberian ham and 2.09 log CFU/g in pressurized Serrano ham. After 60 days at 4 or 8 degrees C, the respective populations in pressurized and control hams were 3.24 and 4.70 log CFU/g for Iberian ham and 2.73 and 5.07 log CFU/g for Serrano ham. The color parameters L* and a* were not influenced by high-pressure treatment, and parameter b* was increased only in Iberian ham. Sensory characteristics of hams were not affected by high-pressure treatment. Treatment of Iberian and Serrano hams at 450 MPa for 10 min significantly reduced the population of L. monocytogenes Scott A without a detrimental effect on the sensory characteristics of the hams.

Inactivation of Escherichia coli O157:H7 in ground beef by single-cycle and multiple-cycle high-pressure treatments.

The effect of single- and multiple-cycle high-pressure treatments on the survival of Escherichia coli CECT 4972, a strain belonging to the O157:H7 serotype, in ground beef was investigated. Beef patties were inoculated with 10(7) CFU/g E. coli O157:H7, and held at 4 degrees C for 20 h before high-pressure treatments. Reduction of the E. coli O157:H7 population by single-cycle treatments at 400 MPa and 12 degrees C ranged from 0.82 log CFU/g for a 1-min cycle to 4.39 log CFU/g for a 20-min cycle. Multiple-cycle treatments were very effective, with four 1-min cycles at 400 MPa and 12 degrees C reducing the E. coli O157:H7 population by 4.38 log CFU/g, and three 5-min cycles by 4.96 log CFU/g. The color parameter L* increased significantly with high-pressure treatments in the interior and the exterior of beef patties, whereas a* decreased in the interior, and b* increased in the exterior-changes that might diminish consumer acceptance of the product. Kramer shear force and energy were generally higher in pressurized than in control ground beef. Maximum values for these texture parameters, which corresponded to tougher patties, were reached after one 10-min cycle in the case of single-cycle treatments or two 5-min cycles in the case of multiple-cycle treatments. High-pressure treatments had no significant effect on Warner-Bratzler shear force.

Inactivation of Salmonella Enteritidis in chicken breast fillets by single-cycle and multiple-cycle high pressure treatments.

The effect of single-cycle and multiple-cycle high hydrostatic pressure (HHP) treatments on the survival of three Salmonella Enteritidis strains in chicken breast fillets was investigated. The surface of fillets was inoculated with a cocktail of three Salmonella strains at approximately 10(7) colony-forming units (CFU)/g, and held at 4 degrees C for 20 hours before HHP treatments. Reduction of Salmonella counts on tryptic soy agar (TSA) by single-cycle treatments at 300 MPa and 12 degrees C ranged from 0.58 log CFU/g for a 0-minute (no dwell time) cycle to 3.35 log CFU/g for a 20-minute cycle, whereas with 400 MPa treatments the decline ranged from 0.93 log CFU/g to more than 5 log CFU/g, respectively. The 4.8 log unit reduction in Salmonella counts on TSA achieved by a 15-minute treatment at 400 MPa should suffice to eliminate the pathogen naturally present in contaminated chicken meat. When plated on Salmonella Shigella agar (SSA), the reduction of Salmonella counts by single-cycle treatments at 300 MPa and 12 degrees C ranged from 0.69 log CFU/g for a 0-minute cycle to 4.21 log CFU/g for a 20-minute cycle, and with 400 MPa treatments from 1.25 log CFU/g to more than 5 log CFU/g, respectively. From the comparison of Salmonella counts on SSA and TSA it was concluded that not only the lethality but also the proportion of injured Salmonella cells increased with the length of HHP treatments. The use of multiple-cycle treatments instead of single-cycle treatments of the same HHP time for the inactivation of Salmonella Enteritidis inoculated on chicken breast fillets showed to be more advantageous at 400 MPa than at 300 MPa. No recovery of injured Salmonella cells was observed when fillets treated at 300 or 400 MPa for 5 minutes were held for 72 hours at 4 degrees C.

Effect of Cheese Water Activity and Carbohydrate Content on the Barotolerance of Listeria monocytogenes Scott A

High-pressure processing is an appropriate technique for improving the microbiological safety of packaged ready-to-eat foods. The effect of high-pressure treatment on Listeria monocytogenes Scott A inoculated into fresh Hispánico-type cheese and ripe Mahón cheese was investigated. A 3.8-log reduction in the counts of L. monocytogenes Scott A in fresh cheese was recorded after 3 min at 400 MPa and 12 degrees C, whereas 18 min under the same conditions was required to obtain a 1-log reduction in ripe cheese. Dry matter values were 48.96% for fresh cheese and 58.79% for ripe cheese, and water activity (aw) values were 0.983 and 0.922, respectively. In dehydrated fresh cheese (58.20% dry matter) in which 5% NaCl was added to achieve a 0.904 aw value, L. monocytogenes Scott A counts were lowered by only 0.4 log after treatment for 10 min at 400 MPa. On the other hand, in a 60:40 mixture of ripe cheese:distilled water with a 0.976 aw value, the reduction under the same conditions was 3.9 log. Within the aw range of 0.945 to 0.965, L. monocytogenes Scott A barotolerance was significantly higher in fresh cheese than in ripe cheese for equivalent aw values. Carbohydrate content was higher in fresh cheese than in ripe cheese. The addition of lactose at a concentration of 5 mg/g to an 85:15 mixture of ripe cheese:distilled water did not influence L. monocytogenes Scott A barotolerance during treatment for 10 min at 400 MPa. Galactose at a concentration of 5 mg/g had a protective effect during high-pressure treatment, and glucose at a concentration of 5 mg/g favored L. monocytogenes Scott A survival during refrigerated storage of pressurized samples at 8 degrees C for 5 days.

Proteolysis and biogenic amine buildup in high-pressure treated ovine milk blue-veined cheese

Penicillium roqueforti plays an important role in the ripening of blue-veined cheeses, mostly due to lactic acid consumption and to its extracellular enzymes. The strong activity of P. roqueforti proteinases may bring about cheese over-ripening. Also, free amino acids at high concentrations serve as substrates for biogenic amine formation. Both facts result in shorter product shelf-life. To prevent over-ripening and buildup of biogenic amines, blue-veined cheeses made from pasteurized ovine milk were high-pressure treated at 400 or 600 MPa after 3, 6, or 9 wk of ripening. Primary and secondary proteolysis, biogenic amines, and sensory characteristics of pressurized and control cheeses were monitored for a 90-d ripening period, followed by a 270-d refrigerated storage period. On d 90, treatments at 400 MPa had lowered counts of lactic acid bacteria and P. roqueforti by less than 2 log units, whereas treatments at 600 MPa had reduced lactic acid bacteria counts by more than 4 log units and P. roqueforti counts by more than 6 log units. No residual α-casein (CN) or κ-CN were detected in control cheese on d 90. Concentrations of β-CN, para-κ-CN, and γ-CN were generally higher in 600 MPa cheeses than in the rest. From d 90 onwards, hydrophilic peptides were at similar levels in pressurized and control cheeses, but hydrophobic peptides and the hydrophobic-to-hydrophilic peptide ratio were at higher levels in pressurized cheeses than in control cheese. Aminopeptidase activity, overall proteolysis, and free amino acid contents were generally higher in control cheese than in pressurized cheeses, particularly if treated at 600 MPa. Tyramine concentration was lower in pressurized cheeses, but tryptamine, phenylethylamine, and putrescine contents were higher in some of the pressurized cheeses than in control cheese. Differences in sensory characteristics between pressurized and control cheeses were generally negligible, with the only exception of treatment at high pressure level (600 MPa) at an early ripening stage (3 wk), which affected biochemical changes and sensory characteristics.

Development of a Potential Probiotic Fresh Cheese Using Two Lactobacillus salivarius Strains Isolated from Human Milk

Cheeses have been proposed as a good alternative to other fermented milk products for the delivery of probiotic bacteria to the consumer. The objective of this study was to assess the survival of two Lactobacillus salivarius strains (CECT5713 and PS2) isolated from human milk during production and storage of fresh cheese for 28 days at 4°C. The effect of such strains on the volatile compounds profile, texture, and other sensorial properties, including an overall consumer acceptance, was also investigated. Both L. salivarius strains remained viable in the cheeses throughout the storage period and a significant reduction in their viable counts was only observed after 21 days. Globally, the addition of the L. salivarius strains did not change significantly neither the chemical composition of the cheese nor texture parameters after the storage period, although cheeses manufactured with L. salivarius CECT5713 presented significantly higher values of hardness. A total of 59 volatile compounds were identified in the headspace of experimental cheeses, and some L. salivarius-associated differences could be identified. All cheeses presented good results of acceptance after the sensory evaluation. Consequently, our results indicated that fresh cheese can be a good vehicle for the two L. salivarius strains analyzed in this study.

Short communication: Antimicrobial effect of lactoferrin and its amidated and pepsin-digested derivatives against Salmonella Enteritidis and Pseudomonas fluorescens

The antimicrobial effect of bovine lactoferrin (LF) and its amidated and pepsin-digested derivatives, at concentrations varying from 0.25 to 20 mg/mL, against 3 Salmonella Enteritidis strains and 3 Pseudomonas fluorescens strains was investigated. Lactoferrin showed its maximum antimicrobial effect at 10 mg/mL against the 3 Salmonella strains, with reductions ranging from 1.3 to 2.0 log units, and the 3 Pseudomonas strains, with reductions ranging from 1.8 to 5.4 log units. In the case of amidated LF, the maximum effect on the 3 Salmonella strains was recorded at 0.25 mg/mL, with reductions in the range of 0.8 to 1.2 log units, whereas it was recorded at 1 mg/mL for the 3 Pseudomonas strains, with reductions in the range of 4.4 to 6.0 log units. Pepsin-digested LF showed its maximum antimicrobial effect at 1 mg/mL against the 3 Salmonella strains, with reductions ranging from 2.6 to 3.4 log units, and at 20 mg/mL against the 3 Pseudomonas strains, with reductions ranging from 4.5 to 5.4 log units. It is worth noting the pronounced effect (reductions exceeding 2.5 log units) of a low (1 mg/mL) concentration of pepsin-digested LF, which is naturally formed in the gastrointestinal tract, on Salmonella and Pseudomonas strains. A highly significant inverse correlation was found between capsule polysaccharide levels of bacterial strains and their lethality in the presence of different concentrations of amidated lactoferrin.

Proteolysis, Texture, and Sensory Characteristics of Serrano Hams from Duroc and Large White Pigs during Dry‐Curing

UNLABELLED Proteolysis, texture, physicochemical, and sensory characteristics of Serrano hams from Duroc and Large White pigs at 5, 7, 9, 12, and 15 m of curing (3 hams per breed per curing time) were investigated. Higher concentrations of some sarcoplasmic and myofibrillar proteins, peptides with molecular mass in the ranges 3700 to 12000 Da, 700 to 2000 Da, and below 450 Da, and total free amino acids were recorded for Large White hams, while hydrophobic peptides were at higher levels in Duroc hams. A 40 kDa peak in the low-ionic-strength soluble protein fraction and 2 peaks of 40 and 45 kDa in the high-ionic-strength soluble protein fraction found only in Duroc hams could be of use in discriminating products from different breeds. Physicochemical and textural characteristics of both types of hams evolved similarly during curing and differences in organoleptic traits between breeds were negligible. Flavor intensity and flavor quality of hams were strongly correlated to curing time, with r(2) values over 0.95 for flavor intensity and over 0.90 for flavor quality, and to the concentration of total free amino acids, with r(2) values over 0.90 for both flavor intensity and flavor quality. The sensory evaluation scores of Duroc hams, at least as high as those of Large White hams, make the production of high-quality Serrano ham from pure breed Duroc pigs feasible. PRACTICAL APPLICATION Duroc breed crosses are advantageous in the production of dry-cured hams, resulting in higher marbling, enhanced flavor and lower processing losses, but the characteristics of pure breed Duroc hams have not been investigated. The similar evolution of the compositional, proteolytic, textural, and sensory characteristics of Serrano hams from Duroc and Large White pigs during dry-curing recorded in the present work makes the production of high-quality dry-cured hams from pure breed Duroc pigs feasible. Three protein peaks found only in Duroc hams can be of use to discriminate products from different breeds.

High-pressure processing decelerates lipolysis and formation of volatile compounds in ovine milk blue-veined cheese.

Enzyme-rich cheeses are prone to over-ripening during refrigerated storage. Blue-veined cheeses fall within this category because of the profuse growth of Penicillium roqueforti in their interior, which results in the production of highly active proteinases, lipases, and other enzymes responsible for the formation of a great number of flavor compounds. To control the excessive formation of free fatty acids (FFA) and volatile compounds, blue-veined cheeses were submitted to high-pressure processing (HPP) at 400 or 600 MPa on d 21, 42, or 63 after manufacture. Cheeses were ripened for 30d at 10°C and 93% relative humidity, followed by 60 d at 5°C, and then held at 3°C until d 360. High-pressure processing influenced the concentrations of acetic acid and short-chain, medium-chain, and long-chain FFA. The effect was dependent on treatment conditions (pressure level and cheese age at the time of treatment). The lowest concentrations of acetic acid and FFA were recorded for cheeses treated at 600 MPa on d 21; these cheeses showed the lowest esterase activity values. Acetic acid and all FFA groups increased during ripening and refrigerated storage. The 102 volatile compounds detected in cheese belonged to 10 chemical groups (5 aldehydes, 12 ketones, 17 alcohols, 12 acids, 35 esters, 9 hydrocarbons, 5 aromatic compounds, 3 nitrogen compounds, 3 terpenes, and 1 sulfur compound). High-pressure processing influenced the levels of 97 individual compounds, whereas 68 individual compounds varied during refrigerated storage. Total concentrations of all groups of volatile compounds were influenced by HPP, but only ketones, acids, esters, and sulfur compounds varied during refrigerated storage. The lowest total concentrations for most groups of volatile compounds were recorded for the cheese pressurized at 600 MPa on d 21. A principal component analysis combining total concentrations of groups of FFA and volatile compounds discriminated cheeses by age and by the pressure level applied to HPP cheeses.

Effect of lactoferrin and its derivatives against gram-positive bacteria in vitro and, combined with high pressure, in chicken breast fillets

The bactericidal activity of lactoferrin (LF), amidated lactoferrin (AMILF), pepsin digested lactoferrin (PDLF), and its activated (ALF) commercial form, against six strains of three gram-positive bacterial species was investigated. Listeria monocytogenes was most sensitive in vitro, Staphylococcus aureus showed a moderate resistance, and Enterococus faecalis was highly resistant to antimicrobials. When chicken breast fillets were inoculated with L. monocytogenes CECT5725 and treated with antimicrobials, reductions were below 0.5 logCFU/ml in all cases. In combination with high pressure (HHP) treatment at 400 MPa for 10 min, antimicrobials showed a slight additional bactericidal effect, always below 1 logCFU/g. Incorporation of antimicrobials 18 h before or 1 h after HHP treatment generally yielded better results than incorporation 1 h before HHP treatment, although reductions remained below 1.5 logCFU/g in all cases. LF and its derivatives showed a limited potential for pathogen control in meat.