Contribution of macrophages to proteolysis and plasmin activity in ewe bulk milk.

A total of 225 bulk sheep milk samples were collected from 5 intensively managed flocks during early, mid, and late lactation to assess the contribution of macrophages to the regulation of the plasmin-plasminogen system. Samples were analyzed for composition, somatic cell counts, milk renneting characteristics, and for plasmin (PL), plasminogen (PG), and plasminogen activators (PA) activities. Isolation of macrophages from milk was performed using a magnetic positive separation and mouse antiovine macrophage antibody; separated cells were lysed by several freeze-thaw cycles, and activity of urokinase PA (u-PA) was determined. Plasmin activity decreased during lactation (42.06 +/- 0.66, early; 31.29 +/- 0.66, mid; 28.19 +/- 0.66 U/mL, late). The reduction in PL activity recorded in the mid and late lactation milk matched the increase in PG:PL ratio. The activity of PA increased throughout lactation; the highest value being recorded in the late lactation milk (260.20 +/- 8.66 U/mL). Counts of isolated and concentrated macrophages were higher in early and mid lactation milk (3.89 +/- 0.08 and 3.98 +/- 0.08 log10 cells/mL, respectively) than in late lactation milk (3.42 +/- 0.08 log10 cells/mL). Stage of lactation did not influence the activity of u-PA detected in isolated macrophages. The activity of u-PA associated with isolated milk macrophages only minimally contributed to total PA activity detected in milk. Proteolytic enzymes, associated with isolated macrophages, act on alpha-casein hydrolysis, as shown by urea-PAGE electrophoresis analysis. Somatic cell counts did not exceed 600,000 cells/mL, and this threshold can be considered a good index of health status of the flock and of the ability of milk to being processed. Our results lend support to the hypothesis that macrophages in ewe bulk milk from healthy flocks only slightly contribute to the activation of the PL-PG system

Probiotic in lamb rennet paste enhances rennet lipolytic activity, and conjugated linoleic acid and linoleic acid content in Pecorino cheese.

Cheeses manufactured using traditional lamb rennet paste, lamb rennet paste containing Lactobacillus acidophilus, and lamb rennet paste containing a mix of Bifidobacterium lactis and Bifidobacterium longum were characterized for the lipolytic pattern during ripening. Lipase activity of lamb rennet paste, lamb rennet containing Lb. acidophilus, and lamb rennet containing a mix of bifidobacteria was measured in sheep milk cream substrate. Rennet paste containing probiotics showed a lipase activity 2-fold greater than that displayed by traditional rennet. Total free fatty acid (FFA) in sheep milk cream was lower in lamb rennet paste (981 microg/g of milk cream) than in lamb rennet containing Lb. acidophilus (1,382.4 microg/g of milk cream) and in lamb rennet containing a mix of bifidobacteria (1,227.5 microg/g of milk cream) according to lipase activity of lamb rennet paste. The major increase of FFA in all cheeses occurred during the first 30 d of ripening with the greatest values being observed for C16:0, C18:0 C18:1. At 60 d of ripening all cheeses showed a reduction in the amount of free fatty acids; in particular, total free fatty acids underwent a decrease of more than 30% from 30 to 60 d in cheeses manufactured using traditional lamb rennet paste, whereas the same parameter decreased 10% in cheeses manufactured using lamb rennet paste containing Lb. acidophilus and cheeses manufactured using lamb rennet paste containing a mix of B. lactis and B. longum. Cheese containing Lb. acidophilus was characterized by the greatest levels of total conjugated linoleic acids (CLA) 9-cis, 11-trans CLA and 9-trans, 11-trans CLA, whereas cheese containing bifidobacteria displayed the greatest levels of free linoleic acid. Rennet pastes containing viable cells of Lb. acidophilus and a mix of B. lactis and B. longum were able to influence the amount of FFA and CLA in Pecorino cheese during ripening

Biochemical patterns in ovine cheese: influence of probiotic strains.

This study was undertaken to evaluate the effect of lamb rennet paste containing probiotic strains on proteolysis, lipolysis, and glycolysis of ovine cheese manufactured with starter cultures. Cheeses included control cheese made with rennet paste, cheese made with rennet paste containing Lactobacillus acidophilus culture (LA-5), and cheese made with rennet paste containing a mix of Bifidobacterium lactis (BB-12) and Bifidobacterium longum (BB-46). Cheeses were sampled at 1, 7, 15, and 30 d of ripening. Starter cultures coupled with probiotics strains contained in rennet paste affected the acidification and coagulation phases leading to the lowest pH in curd and cheese containing probiotics during ripening. As consequence, maturing cheese profiles were different among cheese treatments. Cheeses produced using rennet paste containing probiotics displayed higher percentages of alpha(S1)-I-casein fraction than traditional cheese up to 15 d of ripening. This result could be an outcome of the greater hydrolysis of alpha-casein fraction, attributed to higher activity of the residual chymosin. Further evidence for this trend is available in chromatograms of water-soluble nitrogen fractions, which indicated a more complex profile in cheeses made using lamb paste containing probiotics versus traditional cheese. Differences can be observed for the peaks eluted in the highly hydrophobic zone being higher in cheeses containing probiotics. The proteolytic activity of probiotic bacteria led to increased accumulation of free amino acids. Their concentrations in cheese made with rennet paste containing Lb. acidophilus culture and cheese made with rennet paste containing a mix of B. lactis and B. longum were approximately 2.5 and 3.0 times higher, respectively, than in traditional cheese. Principal component analysis showed a more intense lipolysis in terms of both free fatty acids and conjugated linoleic acid content in probiotic cheeses; in particular, the lipolytic pattern of cheeses containing Lb. acidophilus is distinguished from the other cheeses on the basis of highest content of health-promoting molecules. The metabolic activity of the cheese microflora was also monitored by measuring acetic, lactic, and citric acids during cheese ripening. Cheese acceptability was expressed for color, smell, taste, and texture perceived during cheese consumption. Use of probiotics in trial cheeses did not adversely affect preference or acceptability; in fact, panelists scored probiotic cheeses higher in preference over traditional cheese, albeit not significantly

Effects of Somatic Cell Count and Stage of Lactation on the Plasmin Activity and Cheese-Making Properties of Ewe Milk

The experiment was conducted from March to July 2002 using 5 intensively managed flocks of Southern Italy. In each flock, 2 groups of 50 ewes were created. The groups were designated LSCC (low somatic cell count [SCC]) when their milk SCC was lower than 500,000/mL and HSCC (high SCC) when their milk SCC was higher than 1,000,000/mL. Bulk milk and whey samples were analyzed for fat, total protein, lactose, casein, and whey protein contents. Renneting properties of milk were also determined. Moisture, NaCl, and nitrogen fractions were determined in fresh cheese curds. In addition, plasmin (PL) and plasminogen (PG) activities in milk and cheese were monitored. The proteolytic activity of plasmin by urea-polyacrylamide gel electrophoresis and the white blood cell (WBC) differentials were determined. The HSCC resulted in higher pH values in milk and in higher moisture and lower fat contents in fresh cheese curds. Moreover, a lower recovery of fat and whey proteins was obtained from the HSCC than from the LSCC raw milk. The crude protein and casein contents were higher in the HSCC than in the LSCC curds during early and midlactation; an opposite trend was observed in late lactation. Plasmin and PG activities underwent more marked fluctuations in the LSCC than in the HSCC curds through lactation. The results of this experiment demonstrate that the PL activity in ewe milk is markedly influenced by the SCC, although SCC is not the only parameter for predicting PL and PG evolution in ewe milk. The LSCC milk resulted in a higher proteolytic potential of Canestrato pugliese cheese curds

Influence of ventilation regimen on micro-environment and on ewe welfare and milk yield in summer

The effects of ventilation regimen on air quality, and on the welfare and production performance of thirty-six Comisanaewes were assessed in a 6-week trial conducted during the summer of 2002. Animals were divided into three groups of12, and subjected to the following treatments: low ventilation regimen providing a mean ventilation rate (VR) of 35 m3/hper ewe, split in 30 min ventilation cycles at an air speed of 2 m/s (LOV-30); moderate ventilation regimen (VR = 70m3/h per ewe) split in 30 min ventilation cycles at an air speed of 4 m/s (MOV-30); moderate ventilation regimen (VR =70 m3/h per ewe) split in 60 min ventilation cycles at an air speed of 2 m/s (MOV-60). Air concentrations of microorganisms,dust, and gaseous pollutants were measured twice weekly. Respiration rate (RR) and rectal temperature (RT)were monitored throughout the trial at 0830 and at 1400. Behavioral traits of ewes were recorded twice per week from0900 to 1200 and from 1500 to 1800. Cell-mediated immune response to phytohemagglutinin (PHA) and humoralimmune response to chicken egg albumin were determined. At d 37 ewes were injected with porcine ACTH, and subjectedto blood sampling for evaluation of cortisol concentrations immediately before and 1, 2 and 4 h after ACTH injection.Milk yield was recorded daily. Individual milk samples were analyzed for composition, renneting parameters, somaticcell count (SCC), and bacteriological characteristics. Averages of maximum THI were about 3 points higher in the LOV-30 and the MOV-30 than in the MOV-60 room, whereas no differences emerged in the air concentrations of dust, gaseouspollutants and microorganisms. Significant interactions of treatment x time (P < 0.05) were found for respiration rate,and for the time the ewes spent lying, idling and eating in the afternoon during weeks 2 and 3 of the study period.Significant effects of ventilation regimen x time (P < 0.05) were also observed for milk yield and milk renneting parameters,the LOV-30 ewes giving smaller volumes of milk with a deteriorated coagulating behavior than those of the MOV-60 group during the second half of the trial. No significant differences emerged in ewe immune and endocrine responses.Results show that ventilation regimen had a moderate impact on ewe behavior, physiology and production performance.This experiment suggests that the length of ventilation cycles and air speed, together with ventilation rate, arecritical for efficient ventilation regimens

Consumer acceptance and sensory evaluation of Monti Dauni Meridionali Caciocavallo cheese

Twelve Caciocavallo cheeses were collected from 6 factories (A, B, C, D, E, F) located in the Monti Dauni Meridionali area (Southern Italy) that adopted different protocols for cheese production. A total of 160 consumers were involved in the sensory evaluation of Caciocavallo cheese after 180 d of ripening. Cheese attributes were used to describe the flavor, texture, and appearance of cheeses. The highest scores for the shiny attribute were assigned to cheeses B, C, and E, whereas color intensity was the highest in cheeses B, D, and F. Strength, salty, and piquant attributes were higher in cheeses F and A because of the use of raw milk (F), rennet paste (A), and percentage of salt in the brine (A, F). Consumers perceived a more granular structure during the second half of chewing of Caciocavallo cheese F, as evidenced by the highest value for the grainy attribute. A positive correlation was found between overall flavor and odor intensity and water-soluble nitrogen, low molecular weight peptides, and free fatty acids and between piquant and butyric and caproic acids. A principal components analysis applied to the sensory attributes accounted for 65% of the total variance. The score plot showed that cheeses F and A were located in a well-defined zone of the plot, with cheeses in this zone displaying higher levels of strength, piquant, and salty attributes. The preference test assigned 40% of the preference to Caciocavallo cheese A, 38% to cheese F, 9% to cheese E, 8% to cheese D, and 7% to cheeses B and C. Sensory evaluation of Monti Dauni Meridionali Caciocavallo cheeses is a useful analysis to highlight the principal attributes able to influence consumers' liking that are related to biochemical features of the cheese

Technical note: Rapid method for determination of amino acids in milk.

A rapid method for measurement of amino acids in milk was developed and validated. The method included a first step of milk protein hydrolysis, followed by the derivatization and separation of amino acids by HPLC. Six combinations of hydrolysis agent and temperature-time conditions were compared with a reference method; derivatization procedures as well as HPLC separation were improved. Hydrolysis of milk samples with 6 N HCl at 160 degrees C for 60 min resulted in no significantly differences compared with the reference method but allowed the analysis of a greater number of milk samples in a short time. In addition, this method was characterized by high precision, low repeatability uncertainty, and high accuracy for all amino acids evaluated; the recovery mean value of the single amino acids was 98.38%. The proposed method is, therefore, accurate, simple, rapid, and suitable for large numbers of milk samples

Focusing on fatty acid profile in milk from different species after in vitro digestion

We report the fatty acid profile of raw milk and of the corresponding digested milk from different sources (human milk, formula milk and donkey, bovine, ovine and caprine milk) to gain information on the nutritional quality of different milk sources in infant nutrition.Short chain fatty acids (SC-FA) were higher in bovine and caprine milk, intermediate in ovine and donkey and lower in human and formula milk. Medium chain fatty acids (MC-FA) showed the highest values for bovine and caprine milk and the lowest for donkey and formula milk, whereas long chain fatty acids (LC-FA) were the highest in donkey and formula milk and intermediate in human milk.The percentage distribution of fatty acids liberated after in vitro digestion did not reflect the patterns found in the corresponding milk sources. In particular, MC free fatty acids (MC-FFA) showed the highest and the lowest values in donkey and in formula milk, LC-FFA showed the highest value in human milk. The total FFA was highest in human milk, lowest in formula milk and intermediate in donkey, bovine, ovine, and caprine milk

Consumers' expectations and acceptability for low saturated fat 'salami'

Nutritional properties of meat and meat products are becoming very important in purchasing behaviour, because consumers are even more concerned about healthiness. The present study aimed to examine the influence of health information on the expected and informed acceptability of salami. Traditional salami and two low saturated fat salami produced with partial or total substitution of pork backfat with extra virgin oil were evaluated.; Results: Perceived acceptability was the lowest in salami with total animal fat substitution. In both low saturated fat salami, expected acceptability was significantly higher than perceived acceptability, while in traditional salami it was lower. Consumers completely assimilated their liking in the direction of expectations for salami with partial animal fat substitution, whereas incomplete assimilation was observed for salami with total animal fat substitution. The results also revealed that some sociodemographic characteristics discriminate consumer clusters from each other.; Conclusion: The present study highlights that nutritional information is not enough to satisfy consumers' expectations if the product is not sensorily acceptable. Findings about the relevance of information and consumers' segmentation could have important implications for policy makers and the meat product industry. © 2017 Society of Chemical Industry.; © 2017 Society of Chemical Industry

Milk from different species: Relationship between protein fractions and inflammatory response in infants affected by generalized epilepsy.

The present study was undertaken to evaluate the effect of protein fractions from bovine, caprine, and ovine milk on production of cytokines and reactive oxygen species (ROS) and reactive nitrogen species (RNS) by cultured peripheral blood mononuclear cells (PMBC) from infants with generalized epilepsy. Bovine, caprine, and ovine bulk milks were pasteurized and analyzed for chemical composition. Then, PBMC were isolated from 10 patients with generalized epilepsy (5 males; mean age 33.6±5.4mo). Production of tumor necrosis factor-α (TNF-α), IL-10, IL-6, and IL-1β was studied in cultured PBMC (from infants with epilepsy and controls) stimulated by bovine, caprine, and ovine milk and casein and whey protein fractions, and levels of ROS and RNS were measured in the culture supernatant. The ability of PBMC to secrete cytokines in response to milk and protein fraction stimulation may predict the secretion of soluble factor TNF-α in the bloodstream of challenged patients. Bovine, caprine, and ovine bulk milks induced low-level production of IL-10 by cultured PBMC in at least 50% of cases; the same behavior was observed in both casein and whey protein fractions for all species studied. Bovine and ovine milk and their casein fractions induced production of lower levels of IL-1β in 80% of patients, whereas caprine milk and its casein fraction induced the highest levels in 80% of patients. The amount of IL-6 detected after stimulation of PBMC by milk and its fractions for all species was lower than that of other proinflammatory cytokines. In the bovine, total free radicals were higher in bulk milk and lower in the casein fraction, whereas the whey protein fraction showed an intermediate level; in caprine, ROS/RNS levels were not different among milk fractions, whereas ovine had higher levels for bulk milk and casein than the whey protein fraction. Lower levels of ROS/RNS detected in PBMC cultured with caprine milk fraction could be responsible for the lower levels of TNF-α cytokine in the corresponding fraction. Cytokines might be useful biomarkers to discriminate the effects of foods on the inflammatory response; dietary strategies could help in alleviating the negative effects of epilepsy in infants