Production of low-cholesterol butter with Lacticaseibacillus paracasei immobilized in calcium-alginate beads.

This study focused on the application of three strains of Lacticaseibacillus paracasei to assimilate cholesterol in cream and butter. The strains were enclosed in calcium-alginate beads and incubated in cream at 30 °C for 15 h. Immobilization of lactobacilli cultures in calcium-alginate beads resulted in a 23% reduction in cholesterol (p < 0.05) in cream, whereas a negligible reduction was observed in cream fermented with free cells. Butter with a 44% reduction in cholesterol was produced from fermented cream by L. paracasei L2A21K5 entrapped in alginate beads. No significant (p > 0.05) changes in the fatty acid profile were observed in the low-cholesterol butter, except for a slight but significant increase in n-3 fatty acids (p < 0.05). In addition, the indices of atherogenicity and thrombogenicity were significantly reduced in the low-cholesterol butter (p < 0.05). Panelists rated the low-cholesterol butter as good in appearance, consistency, and flavor.

Short communication: Latin-style fresh cheese enhances lactic acid bacteria survival but not Listeria monocytogenes resistance under in vitro simulated gastrointestinal conditions.

Different studies in humans have provided evidence about the health benefits of probiotics. However, most probiotic strains do not maintain good viability in the harsh conditions of the gastrointestinal tract (GIT). In the present study, Latin-style fresh cheese produced with potential probiotic bacteria was tested to evaluate this cheese type as a food carrier for the delivery of viable microorganisms after exposure to simulated GIT conditions. The resistance of 28 lactic acid bacteria (LAB) strains and Listeria monocytogenes upon exposure to acidic conditions (pH 2.5) and bile and pancreatic enzymes (0.3% of bile salts and 0.1% of pancreatin) was evaluated in vitro. When compared with fresh cultures, fresh cheese greatly improved LAB survival to simulated GIT conditions, as no loss of viability was observed in either acidic conditions (pH 2.5) or bile salts and pancreatin environment over a 3-h period. In opposition, L. monocytogenes did not survive after 1h under acidic conditions. These data demonstrated that Latin-style fresh cheese could play an important role in probiotic protection against gastrointestinal juices, enhancing delivery within the gut and thereby maximizing potential health benefits of LAB.

Control of Listeria monocytogenes in fresh cheese using protective lactic acid bacteria.

In the past years, there has been a particular focus on the application of bacteriocins produced by lactic acid bacteria (LAB) in controlling the growth of pathogenic bacteria in foods. The aim of this study was to select LAB strains with antimicrobial activity, previously isolated from a traditional Azorean artisanal cheese (Pico cheese), in order to identify those with the greatest potential in reducing Listeria monocytogenes in fresh cheese. Eight bacteriocin producer strains identified as Lactococcus lactis (1) and Enterococcus faecalis (7) were tested. In general, the bacteriocin-producing strains presented a moderate growth in fresh cheese at refrigeration temperatures (4 °C), increasing one log count in three days. They exhibited slow acidification capacity, despite the increased production of lactic acid displayed by some strains after 24h. Bacteriocin activity was only detected in the whey of fresh cheese inoculated with two Enterococcus strains, but all cheeses made with bacteriocin-producing strains inhibited L. monocytogenes growth in the agar diffusion bioassay. No significant differences were found in overall sensory evaluation made by a non-trained panel of 50-52 tasters using the isolates as adjunct culture in fresh cheese, with the exception of one Enterococcus strain. To test the effect of in situ bacteriocin production against L. monocytogenes, fresh cheese was made from pasteurized cows' milk inoculated with bacteriocin-producing LAB and artificially contaminated with approximately 10(6) CFU/mL of L. monocytogenes. The numbers of L. monocytogenes were monitored during storage of fresh cheese at refrigeration temperature (4 °C) for up to 15 days. All strains controlled the growth of L. monocytogenes, although some Enterococcus were more effective in reducing the pathogen counts. After 7 days, this reduction was of approximately 4 log units compared to the positive control. In comparison, an increase of 4 log CFU/mL in pathogen numbers was detected over the same period, in the absence of bacteriocin-producing LAB. The combination of two bacteriocin producing Enterococcus sp. optimized the reduction of L. monocytogenes counts in fresh cheese, reducing by approximately 5 log units after 7 days. The present work demonstrates that using bacteriocin-producing strains in the manufacture of fresh cheese might contribute to preventing the growth of undesirable pathogenic bacteria such as L. monocytogenes. A blend of two strains demonstrated great potential as a protective culture for the cheese making process.

Effect of corn supplementation of grass finishing of Holstein bulls on fatty acid composition of meat lipids.

Finishing Holstein young bulls exclusively on pasture generally results in very lean carcass and meat, but corn supplementation is expected to simultaneously improve carcass traits and intramuscular lipids (IML). The expected increase in IML would allow for a larger 18:2c9,t11 (CLA) deposition in meat without affecting the n-3 PUFA present in LM phospholipids (PL). Holstein bulls (n = 33) with initial BW of 423 ± 52.4 kg reared exclusively on pasture were assigned to 1 of 3 finishing period (85 d) diets: finished exclusively on pasture (P0) or finished on pasture and individually supplemented with 4 (P4) or 8 kg/d (P8) of ground corn. Final BW (546 ± 56.3 kg) was not affected (P > 0.05) by corn supplementation, but ADG increased (P < 0.01) with the increasing corn supplementation level from 1.23 kg/d for P0 to 1.44 kg/d for P4 and to 1.67 kg/d for P8. Subcutaneous fat depth in P0 bulls was 0.8 mm and increased (P < 0.001) in both P4 (2.9 mm) and P8 (2.7 mm) bulls, but no difference (P = 0.73) was observed between P4 and P8 bulls. Similarly, the IML increased with corn supplementation, from 1.84 g/100 g muscle in P0 to 2.96 in P4 and to 3.24 in P8, but no difference (P = 0.55) was found between P4 and P8 bulls. Corn supplementation decreased (P < 0.01) 18:1t11 in neutral lipids (NL) but not 18:2c9,t11 (P > 0.34). The 18:1t10 (mg/g total NL fatty acid [FA] ± SEM) were 2.5 ± 0.13 in P0, 5.5 ± 1.68 in P4, and 14.8 ± 3.18 in P8 bulls, being greater in P8 compared with P4 (P = 0.02). Total FA in muscle PL and SFA were unaffected, but increasing corn supplementation resulted (P < 0.001) in an increase of 18:2n-6 in PL by replacement of mostly the 18:1c9 and 18:3n-3. Notably, the total number of cis double bonds present in FA of PL remained constant (P = 0.74) with corn supplementation. Compared with P0, corn supplementation maintained (P > 0.05) the high n-3 PUFA content in meat (mg/100 g meat) and increased the 18:2c9,t11 (P = 0.028) and 18:1c9 (P < 0.001). However, increasing corn supplementation from 4 to 8 kg/d increased the 18:1t10 (P = 0.031) and had no effect on 18:2c9,t11. Therefore, supplementing grass-finished Holstein bulls with moderate amounts of ground corn (4 kg/d) increased carcass fat cover and IML, maintained n-3 PUFA, and increased 18:2c9,t11 content in meat, whereas greater corn supplementation (8 kg/d; P8) resulted in no further improvements.

Consumption of high energy maize diets is associated with increased soluble collagen in muscle of Holstein bulls.

The aim was to investigate whether energy supplementation of gazing bulls can influence collagen concentration and solubility in three different muscles: longissimus dorsi (LD), semitendinosus (ST) and supraspinatus (SS). Thirty three Holstein bulls aged 15 months and reared on grass, were randomly assigned to three treatment groups for 85 days: fed grass ad libitum (G1-control); fed grass ad libitum and supplemented with 4 kg day(-1) of ground maize (G2); fed grass ad libitum and supplemented with 8 kg day(-1) of ground maize (G3). Total collagen content did not differ (P>0.05) among treatments, however, soluble (heat-labile) collagen was significantly (P<0.001) higher in meat from G3 (34%, 23% and 25% for LD, ST and SS, respectively), compared with G1-control (24%, 18% and 17% in LD, ST and SS, respectively). Overall, these results suggest that meat tenderness of grazing bulls may be improved by supplementing with ground maize (8 kg day(-1)).

Rumen biohydrogenation-derived fatty acids in milk fat from grazing dairy cows supplemented with rapeseed, sunflower, or linseed oils.

The effects of supplementation with rapeseed, sunflower, and linseed oils (0.5 kg/d; good sources of oleic, linoleic, and linolenic acids, respectively) on milk responses and milk fat fatty acid (FA) profile, with special emphasis on rumen-derived biohydrogenation intermediates (BI), were evaluated in a replicated 4 x 4 Latin square study using 16 grazing dairy cows. The dietary treatments were 1) control diet: 20-h access to grazing pasture supplemented with 5 kg/d of corn-based concentrate mixture (96% corn; CC); 2) RO diet: 20-h access to grazing supplemented with 4.5 kg/d of CC and 0.5 kg of rapeseed oil; 3) SO diet: 20-h access to grazing supplemented with 4.5 kg/d of CC and 0.5 kg of sunflower oil; and 4) LO diet: 20-h access to grazing supplemented with 4.5 kg/d of CC and 0.5 kg of linseed oil. Milk fatty acids were converted to methyl esters and analyzed by gas-liquid chromatography and silver-ion HPLC. Dietary treatments had no effect on milk production or on milk protein content and milk protein production. Supplementation with rapeseed and sunflower oils lowered milk fat content and milk fat production, but linseed oil had no effect. Inclusion of dietary vegetable oils promoted lower concentrations of short-chain (including 4:0) and medium-chain FA (including odd- and branched-chain FA) and 18:3n-3, and higher concentrations of C(18) FA (including stearic and oleic acids). The BI concentration was higher with the dietary inclusion of vegetable oils, although the magnitude of the concentration and its pattern differed between oils. The RO treatment resulted in moderate increases in BI, including trans 18:1 isomers and 18:2 trans-7,cis-9, but failed to increase 18:1 trans-11 and 18:2 cis-9,trans-11. Sunflower oil supplementation resulted in the highest concentrations of the 18:1 trans-10, 18:1 cis-12, and 18:2 trans-10,trans-12 isomers. Concentrations of 18:1 trans-11 and 18:2 cis-9,trans-11 were higher than with the control and RO treatments but were similar to the LO treatment. Concentration of BI in milk fat was maximal with LO, having the highest concentrations of some 18:1 isomers (i.e., trans-13/14, trans-15, cis-15, cis-16), most of the nonconjugated 18:2 isomers (i.e., trans-11,trans-15, trans-11,cis-15, cis-9,cis-15, and cis-12,cis-15), and conjugated 18:2 isomers (i.e., trans-11,cis-13, cis-12,trans-14, trans-11,trans-13, trans-12,trans-14, and trans-9,trans-11), and all conjugated 18:3 isomers. The LO treatment induced the highest amount and diversity of BI without decreasing milk fat concentration, as the RO and SO treatments had, suggesting that the BI associated with 18:3n-3 intake may not be the major contributors to inhibition of mammary milk fat synthesis.

Effects of grass silage and soybean meal supplementation on milk production and milk fatty acid profiles of grazing dairy cows.

The effects of supplementation with grass silage and replacement of some corn in the concentrate with soybean meal (SBM) on milk production, and milk fatty acid (FA) profiles were evaluated in a replicated 4 x 4 Latin square study using 16 dairy cows grazing pasture composed of ryegrass, Kentucky bluegrass, and white clover. Each experimental period lasted for 3 wk. The 4 dietary treatments were PC, 20 h of access to grazing pasture, supplemented with 6 kg/d of corn-based concentrate mixture (96% corn; C); PCSB, 20 h of access to grazing pasture, supplemented with 6 kg/d of corn- and SBM-based concentrate mixture (78% corn and 18% SBM; CSB); SC, 7 h of access to grazing pasture during the day and 13 h of ad libitum access to grass silage at night, supplemented with 6 kg/d of C concentrate; and SCSB, 7 h of access to grazing pasture during the day and 13 h of ad libitum access to grass silage at night, supplemented with 6 kg/d of CSB concentrate. The concentrate mixtures were offered twice each day in the milking parlor and were consumed completely. Grass silage supplementation reduced dietary crude protein and concentration of total sugars, and dietary SBM inclusion increased dietary crude protein concentration and decreased dietary starch concentration. Milk yield and energy-corrected milk were increased by SBM supplementation of cows with access to grass silage. Milk protein concentration was lower in cows offered grass silage, regardless of whether SBM was fed. Dietary SBM inclusion tended to increase milk fat concentration. Plasma urea N was reduced by silage feeding and increased by SBM supplementation. Supplementation with grass silage overnight could represent a useful strategy for periods of lower pasture availability. Dietary inclusion of SBM in solely grazing cows had no effects on milk production and composition, exacerbated the inefficient capture of dietary N, and increased diet cost. Grass silage supplementation affected milk FA profiles, increasing both the FA derived from de novo synthesis and those derived from rumen microbial biomass, and decreasing the sum of C18 FA (mostly derived from diet or from mobilization of adipose tissue reserves). Milk fat concentrations of conjugated linoleic acid cis-9, trans-11, vaccenic acid (18:1 trans-11), and linolenic acid (18:3n-3) were unaffected by grass silage supplementation, suggesting that partial replacement of pasture by unwilted grass silage does not compromise the dietary quality of milk fat for humans.

Seasonal variations in the developmental competence of bovine oocytes matured in vitro.

Ovaries were collected over a period of two years from heifers slaughtered at under 30 months of age and used to harvest 1757 oocytes. After in vitro maturation, fertilisation and culture, the proportions of oocytes and cleaved embryos that developed to blastocysts were significantly higher (P<0.01) in the autumn, from September to November, than in the spring, from March to May. In contrast, embryo development, as assessed by oocytes that developed to eight or more cells and blastocysts, was lowest (P<0.01) in the spring. These results were consistent during the two-year study, indicating a seasonal fluctuation in oocyte competence.