Adhesion of dairy propionibacteria to intestinal epithelial tissue in vitro and in vivo

Adhesion to the intestinal mucosa is a desirable property for probiotic microorganisms and has been related to many of their health benefits. In the present study, 24 dairy Propionibacterium strains were assessed with regard to their hydrophobic characteristics and their autoaggregation and hemagglutination abilities, since these traits have been shown to be indicative of adherence in other microorganisms. Six strains were further tested for their capacity to adhere to ileal epithelial cells in vitro and in vivo. The results of the study showed that propionibacteria were highly hydrophilic, and hemagglutination and autoaggregation were properties not commonly found among these microorganisms. No relationship was found between surface characteristics and adhesion ability, since hemagglutinating, autoaggregating, and nonautoaggregating bacteria were able to adhere to intestinal cells both in vitro and in vivo. Microscopic examination revealed that autoaggregating cells adhered in clusters, with adhesion being mediated by only a few bacteria, whereas the hemagglutinating and nonautoaggregating strains adhered individually or in small groups making contact with each epithelial cell with the entire bacterial surface. The in vitro assessment of adhesion was a good indication of the in vivo association of propionibacteria with the intestinal epithelium. Therefore, the in vitro method presented here should be valuable in screening routinely adhesive properties of propionibacteria for probiotic purposes. The adhesion ability of dairy propionibacteria would prolong their maintenance in the gut and increase the duration of their provision of beneficial effects in the host, supporting the potential of Propionibacterium in the development of new probiotic products.     

Inability of dairy propionibacteria to grow in milk from low inocula

Growth of propionibacteria in complex media was independent of the initial number of cells; in contrast, growth of propionibacteria in milk and whey did not occur if the initial level of cells was < 10(6) cfu/ml. Addition of vitamins, minerals or complex nitrogen sources to the milk or whey, or incubation under anaerobic conditions had no effect on the lack of growth. Addition of freeze-dried whey, prepared from skim milk reconstituted from powder, to a complex medium prevented growth from low inocula in the complex medium, demonstrating the presence of an inhibitor or inhibitors in the whey. The inhibitor(s) was heat stable, had a low molecular mass and retained its activity for at least 4 weeks at 20 degrees C. Pregrowth of some lactic acid bacteria, used as starter cultures in Swiss-type cheese manufacture, in milk for 2 weeks at 20 degrees C removed the inhibition, which explains how propionibacteria develop in Swiss-type cheese from low numbers even though they are inhibited in milk.     

Effect of feeding propionibacteria on milk production by early lactation dairy cows

This experiment was conducted to determine the effect of a direct-fed microbial agent, Propionibacterium strain P169 (P169), on rumen fermentation, milk production, and health of periparturient and early-lactation dairy cows. Starting 2 wk before anticipated calving, cows were divided into 2 groups and fed a control diet or the control diet plus 6 × 10¹¹ cfu/d of P169. Cows were changed to a lactation diet at calving, and treatments continued until 119 d in milk. Rumen fluid samples were taken about 1 wk before calving, and at 1 and 14 wk after calving. Cows fed P169 had lower concentrations of acetate (mol/100 mol of total volatile fatty acids) at all time points, greater concentrations of propionate on the first and last sampling points, and greater concentrations of butyrate on the first 2 time points. Concentrations of glucose in plasma and milk and plasma concentrations of β-hydroxybutyrate were not affected by treatment. Cows fed P169 had greater concentrations of plasma nonesterified fatty acids on d 7 of lactation. The high nonesterified fatty acids at that time point was probably related to the high production of milk during that period by cows fed the additive. Cows fed P169 during the first 17 wk of lactation produced similar amounts of milk (44.9 vs. 45.3 kg/d, treatment vs. control) with similar composition as cows fed the control diet. Calculated net energy use for milk production, maintenance, and body weight change was similar between treatments, but cows fed the P169 consumed less dry matter (22.5 vs. 23.5 kg/d), which resulted in a 4.4% increase in energetic efficiency.     

Influence of dairy practices on the capacity of probiotic bacteria to overcome simulated gastric digestion

The aim of this study was to evaluate the influence of microbiological and technological factors on the viability and functionality of probiotic Lactobacillus strains. In particular, the influence of harvesting time, food matrix, refrigerated storage, time of inoculation and refrigerated storage in fermented milk on the resistance to simulated gastric digestion (RSGD) was evaluated. Moreover, strain resistance to simulated gastric digestion was compared to human gastric fluid. Results showed that the variables studied affected, in a different way and in a strain‐dependent manner, the RSGD. No direct relation was observed between cell viability and RSGD.     

Effects of feeding two levels of propionibacteria to dairy cows on plasma hormones and metabolites

To determine the effect of feeding propionibacteria on metabolic indicators during lactation, multiparous and primiparous Holstein cows were fed one of three dietary treatments in a 2 x 3 factorial design from 2 weeks prepartum to 30 weeks post partum: (1) Control (primiparous n=5, multiparous n=8) fed a total mixed ration (TMR); (2) high-dose group (primiparous n=6, multiparous n=5) fed TMR plus 6 x 10 (11) cfu/head daily (high-dose P169) of propionibacterium strain P169; or (3) low-dose group (primiparous n=8, multiparous n=6) fed TMR plus 6 x 10(10) cfu/head daily (low-dose P169) of P169. Blood samples were collected weekly and analysed for plasma concentrations of glucose, insulin, insulin-like growth factor-I (IGF-I), leptin, nonesterified fatty acids (NEFA) and cholesterol. Between weeks 25 and 30, all groups received bovine somatotropin (bST) every 2 weeks. Low-dose P169 multiparous cows had lower (P<0.05) plasma insulin and glucose concentrations than high-dose P169 multiparous cows, whereas high-dose P169 primiparous cows had lower glucose but greater insulin concentartions than low-dose P169 primiparous cows (P<0.05). Plasma insulinratioglucose molar ratios were 13-18% lower (P<0.05) in low-dose P169 cows than in control or high-dose P169 cows. Plasma IGF-I, NEFA and leptin levels did not differ among diet groups between weeks 1 and 25. Low-dose P169 multiparous cows had 25% greater plasma cholesterol levels than high-dose P169 and control multiparous cows, but cholesterol levels in primiparous cows did not differ. During bST treatment, high-dose P169 multiparous cows and low-dose P169 primiparous cows had lower IGF-I levels than their respective controls and, regardless of parity, high-dose P169 cows had greater NEFA than control cows. Although supplemental feeding of P169 altered plasma hormones and metabolites, the particular effects were dependent on dose of P169 and parity of cows.     

Effects of propionibacteria and yeast culture fed to steers on nutrient intake and site and extent of digestion

The effects of feeding Propionibacterium strain P169 (P169), yeast culture (XPY), and their combination on nutrient intake, site and extent of digestion, and ruminal kinetics were evaluated in a completely randomized experimental design. Ruminally and duodenally cannulated Angus × Hereford steers (n = 12) were assigned to 1 of 4 treatments in each of 2 periods: 1) control, fed a sorghum silage-based total mixed ration; 2) P169, fed the control plus P169 (6 × 10¹¹ cfu/steer per d); 3) XPY, fed the control plus XPY (56 g/steer per d); and 4) P169 + XPY, fed the control plus P169 and XPY (at 6 × 10¹¹ cfu/steer per d and 56 g/steer per d, respectively). Each period lasted 21 d; d 1 to 15 were used for diet adaptation and d 16 to 21 were used for fecal, duodenal, ruminal, and blood sample collection. Steers were individually housed and fed. Feeding XPY tended to decrease intake of organic matter, acid detergent fiber, and N, and decreased intake of neutral detergent fiber. However, feeding XPY alone tended to increase total tract digestibility of organic matter, N, neutral detergent fiber, and acid detergent fiber. Ruminal digestibility, duodenal flow, microbial N synthesis, microbial efficiency, and fluid and particulate passage rates were not affected by dietary treatments. Feeding P169 tended to decrease molar proportion of acetate, increased molar proportion of propionate (by 9.7%), and tended to decrease acetate:propionate ratio compared with control steers. No other effects of XPY or P169 on ruminal fermentation were observed. Plasma glucose and insulin concentrations were not affected by dietary treatment. Our results suggest that feeding P169 alters ruminal metabolism toward increased propionate without affecting feed intake or ruminal kinetics, whereas feeding XPY alone tended to increase total tract digestibilities of nutrients.     

Effects of feeding propionibacteria to dairy cows on milk yield, milk components, and reproduction

Two weeks before parturition, 38 Holstein primiparous and multiparous cows were assigned to 1 of 3 treatment groups: control animals (n = 13) received regular total mixed rations (TMR), the low-dose group (n = 14) received the control TMR plus 6 x 10(10) cfu/cow of Propionibacterium strain P169 (P169), and the high-dose group (n = 11) received the control TMR plus 6 x 10(11) cfu/cow of P169 from -2 to 30 wk postpartum. Weekly milk samples were analyzed for percentage of milk fat, protein, lactose, and SNF, milk urea nitrogen, and somatic cell counts. Daily milk production expressed as 4% fat-corrected milk was affected by treatment and week x parity. High-dose and low-dose P169-treated cows exhibited 7.1 and 8.5% increases above controls in daily 4% fat-corrected milk, respectively. Treatment x parity and week significantly influenced percentage of milk fat, lactose, and protein, whereas treatment x parity and treatment x week influenced SNF. Ruminal propionate levels were influenced by treatment such that high-dose P169 cows had greater molar percentage of propionate than did low-dose P169 and control cows. Change in body weight postpartum was influenced by week x parity and treatment x parity such that high-dose and low-dose P169 multiparous cows exhibited a more rapid recovery of wk-1 body weight than did control multiparous cows. There was no treatment, parity, or interaction on days to first postpartum ovulation or on estrous behavior at 45 and 90 d postpartum. We concluded that P169 might have potential as an effective direct-fed microorganism to increase milk production in dairy cows.     

Pregnancy per artificial insemination in lactating dairy cows subjected to 2 different intervals from presynchronization to initiation of Ovsynch protocol

A protocol for presynchronization of ovarian status with 2 injections of PGF_2α given 14 d apart, with the last PGF_2α injection given 12 or 14 d before Ovsynch increases pregnancy per artificial insemination (P/AI) in dairy cows. We determined the efficacy of reducing the interval from the last PGF_2α injection (500 μg of cloprostenol) of presynchronization to initiation of Ovsynch on response to treatment and P/AI. Lactating dairy cows were assigned to an Ovsynch protocol, with the initial injection of GnRH given either 9 (PRE-9; n = 135) or 12 d (PRE-12; n = 135) after the second PGF_2α injection of presynchronization. The Ovsynch protocol consisted of 2 injections of 100 μg of GnRH given 9 d apart and 1 injection of PGF_2α given 7 d after the initial GnRH injection, and cows were subjected to timed artificial insemination (TAI; 70 ± 3.5 DIM) approximately 16 h after the second GnRH injection. Body condition score (1–5 scale) was recorded at TAI. Blood samples were taken for progesterone determination at the PGF_2α injection of Ovsynch, at TAI, and at 11 d after TAI. Ultrasonographic examinations were done in all cows at the second PGF_2α injection of presynchronization, initial GnRH injection, PGF_2α injection of Ovsynch, at TAI, and 24 h after TAI for cyclicity status and ovarian responses to treatments, and at 32 and 60 d after TAI for confirmation of pregnancy. Overall, 29 cows (10.7%) were determined acyclic or cystic and excluded from the study. The percentage of cows responding to initial GnRH injection (62.2 vs. 61.5%) did not differ between PRE-9 and PRE-12 but more cows in the PRE-9 group failed to respond to PGF_2α treatment of Ovsynch compared with PRE-12 (22.7 vs. 10.7%). Body condition score at TAI (2.9 ± 0.02) and mean ovulatory follicle diameter (16.4 ± 0.2 mm) were not different between treatments. Overall P/AI at 32 d was reduced in PRE-9 (33.6%) compared with PRE-12 (44.3%) but pregnancy losses (5.0 vs. 3.7%) did not differ between treatments. Primiparous cows in the PRE-12 group had higher mean progesterone concentration 11 d after TAI and greater P/AI 32 after TAI than primiparous cows in the PRE-9 group (6.4 ± 0.5 vs. 4.6 ± 0.5 ng/mL and 55.8 vs. 30.0%, respectively). In conclusion, reducing the interval from the last PGF_2α injection of the presynchronization treatment to initiation of Ovsynch (from 12 to 9 d) did not affect ovulatory response to initial GnRH injection but reduced response to PGF_2α injection of Ovsynch and P/AI at 32 and 60 d after TAI. The reduction in P/AI was particularly evident in primiparous cows of the PRE-9 group.     

Effects of simulated gastric and intestinal digestion on chitooligosaccharides in two in vitro models

Chitooligosaccharides (COS) with different degree of polymerisation (DP) have different physiological activities such as anti-tumour and anti-hyperlipidemia activities. However, the digestive process might lead to the change of DP of COS as well as cause the change of physiological activities in vivo. In this study, two in vitro digestion models (static and dynamic) were used to investigate digestion behaviours of COS and the influencing factors during digestion with or without food matrix. The results showed that COS with DP 2–5 were indigestible and COS with DP 6–10 degraded with time during gastric and intestinal digestion. Pepsin, pancreatin and lipase were the main factors leading to degradation. Food matrix could protect COS with DP 6–10 from degradation during gastric digestion. However, the degradation of COS with DP 6–10 accelerated and the degradation of COS with DP 5 occurred during intestinal digestion with food matrix due to pancreatin, lipase and bile salt.     

Growth and nutrient digestion by dairy heifers fed semipurified diets

Holstein heifers, which weighed an average of 154 kg, were randomly assigned to three dietary treatments in a split plot in time design. Differences in diets were fiber source: solka floc, corn cobs, and corn silage. The major source of added protein was corn gluten meal. Total collection metabolism periods were the final 7 d of each of two 30-d periods. Several heifers had simple digestive disturbances, which appeared to be related to low ruminal pH, while consuming the solka floc diet. Average daily gains were .78, .83, and 1.02 kg/d for treatments solka floc, corn cobs, and corn silage, respectively. Gains were within the range of acceptable growth standards and were higher than reports in the literature for semipurified and purified diets. Protein in corn gluten meal appeared to be utilized efficiently by the heifers for growth. The solka floc and corn cob diets are acceptable for growing dairy heifers where a low mineral content is desired but normal growth rates need to be maintained. The solka floc diet might be improved by including a buffer to help stabilize rumen pH.     

Nutrient demand interacts with grass particle length to affect digestion responses and chewing activity in dairy cows

Effects of grass particle length on dry matter intake (DMI), milk production, ruminal fermentation and pool sizes, digestion and passage kinetics, and chewing activity and the relationship of these effects with preliminary DMI (pDMI) were evaluated using 15 ruminally and duodenally cannulated Holstein cows in a crossover design with a 14-d preliminary period and two 18-d treatment periods. During the preliminary period, pDMI of individual cows ranged from 22.6 to 29.8 kg/d (mean=25.8 kg/d) and 3.5% fat-corrected milk yield ranged from 29.2 to 56.9 kg/d (mean=41.9 kg/d). Experimental treatments were diets containing orchardgrass silage chopped to either (a) 19-mm (long) or (b) 10-mm (short) theoretical length of cut as the sole forage. Grass silages contained approximately 46% neutral detergent fiber (NDF); diets contained 50% forage, 23% forage NDF, and 28% total NDF. Preliminary DMI, an index of nutrient demand, was determined during the last 4 d of the preliminary period when cows were fed a common diet and used as a covariate. Main effects of grass particle length and their interaction with pDMI were tested by ANOVA. Grass particle length and its interaction with pDMI did not affect milk yield, milk composition, or rumen pH. Long particle length tended to decrease DMI compared with short particle length, which might have been limited by rumen fill or chewing time, or both. Passage rates of feed fractions did not differ between long and short particle lengths and were not related to level of intake. As pDMI increased, long particles decreased ruminal digestion rate of potentially digestible NDF at a faster rate than short particles. As a result, long particles decreased or tended to decrease rates of ruminal turnover for NDF, organic matter, and dry matter and increased their rumen pools compared with short particles for cows with high pDMI. Long particles increased eating time, which affected cows with high intake to the greatest extent, and total chewing time compared with short particles. As intake increased, ruminal digestion (kg/d) and digestibility (%) of starch decreased, rumen pool size of starch increased, and postruminal digestion and digestibility of starch increased quadratically. When grass silage was the only source of forage in the diet, increasing chop length from 10 to 19 mm tended to decrease DMI but did not negatively affect productivity of cows, which were fed adequate fiber.     

Stability of polyphenols in chokeberry (Aronia melanocarpa) subjected to in vitro gastric and pancreatic digestion

Berries and red fruits are dietary sources of polyphenols with reported health benefits. As part of the diet, polyphenols are ingested as complex mixtures immersed in a food matrix which is digested in the gut. Epithelial cells lining the gut are regularly exposed to these digested mixtures. To understand the effects of dietary polyphenols on human gut health it is essential to determine their stability and fate in the lumen. In this work, we investigated the effects of an in vitro gastric and pancreatic digestion on the stability and composition of the major polyphenols in chokeberry juice. Digestion was carried out with a mixture of pepsin-HCl for 2 h, followed by a 2 h incubation with pancreatin and bile salts at 37 °C with shaking, in the absence of light and under N₂. After digestion, the chokeberry samples were acidified, filtered and HPLC-DAD/HPLC–MS–MS analysed to determine the content of total soluble recovered phenolics. Gastric digestion had no substantial effect on any of the major phenolic compounds in chokeberry, namely anthocyanins, flavan-3-ols, flavonols and caffeic acid derivatives. However, these compounds were significantly altered during the pancreatic digestion and this effect was more marked for anthocyanins (approximately 43% was lost during the 2 h treatment with pancreatin). Flavonols and flavan-3-ols decreased by 26% and 19%, respectively. Neochlorogenic acid decreased by 28% whereas chlorogenic acid was increased by 24%. In vitro digestion of standard phenolic compounds, representing each of the groups of phenolics in chokeberry, confirmed some of the observed changes. Interactions with the digestive enzymes were not responsible for the observed losses which were mostly due to the chemical conditions during pancreatic digestion. Our results, in accordance with previously published results, show that dietary polyphenols are highly sensitive to the mild alkaline conditions in the small intestine and that a good proportion of these compounds can be transformed into other unknown and/or undetected structural forms with different chemical properties and, consequently, different bioaccessibility, bioavailability and biological activity.     

Aggregation and hydrophobicity properties of 6 dairy propionibacteria strains isolated from homemade Turkish cheeses

In the present study, 6 dairy Propionibacterium strains were assessed with regard to their hydrophobic characteristics and their autoaggregation and coaggregation abilities since these traits have been shown to be indicative of adherence in other microorganisms. Aggregation assays and bacterial adhesion to hydrocarbons demonstrated significant differences in cell surface properties among the tested propionibacteria strains. Almost all strains appeared relatively hydrophilic, which showed low affinity for p-xylene. Four of the tested strains showed the strong adhesion to ethyl acetate, a basic solvent, in comparison with microbial adhesion to chloroform, an acidic solvent, which demonstrated the particularity of propionibacteria to have an important electron donor and acidic character. Also, these strains simultaneously showed affinity to 3 hydrocarbons, suggesting a high complexity of the cell surface. All propionibacteria strains tested showed autoaggregation and coaggregation ability with the Escherichia coli ATTC 11229, but the results were strain-specific and dependent on incubation conditions. Anaerobic incubation conditions were determined as the best condition for aggregation abilities of propionibacteria strains. A relationship was obtained between aggregation abilities (auto- and coaggregation) and a correlation between adhesion to hydrocarbon (chloroform) and autoaggregation was possible. Our results indicate that the ability to autoaggregation together with cell surface hydrophobicity and coaggregation abilities with E. coli strain can be used for preliminary screening in order to identify potentially probiotic bacteria suitable for human or animal use. PRACTICAL APPLICATION: Autoaggregation, cell surface hydrophobicity, and coaggregation abilities with E. coli of the selected dairy propionibacteria strains could be used as probiotic in foods after in vivo studies.     

Effects of presynchronization and length of proestrus on fertility of grazing dairy cows subjected to a 5-day timed artificial insemination protocol

The objectives were to compare the effects of 2 methods of presynchronization and 2 lengths of proestrus on fertility of grazing dairy cows subjected to a 5-d timed artificial insemination (AI) protocol at initiation of breeding season. Lactating dairy cows (n=1,754) from 3 seasonal grazing farms were blocked within farm by breed, parity, and days in milk (DIM). Study d 0 was considered the day of AI of cows in COS72 (72h of proestrus). Within each block, cows were randomly assigned to 1 of 2 presynchronization treatments: a PGF(2α)-based program, Presynch, consisting of 2 injections of PGF(2α) administered on d -32 and -18, or a PGF(2α)-GnRH-based program, Double-Ovsynch (DO), consisting of GnRH on d -25, PGF(2α) on d -18, and GnRH on d -15. Within each of the 2 presynchronization treatments, cows were randomly assigned to 1 of 2 lengths of proestrus within the 5-d timed AI protocol, consisting of GnRH on d -8, PGF(2α) on d -3 and -2, and GnRH+AI at either 58 h (COS58) or 72 h (COS72) after the d -3 PGF(2α) injection. Ovaries were scanned by ultrasonography twice, on d -42 and -32, to determine estrous cyclicity before enrollment in the study. Blood was sampled and analyzed for concentrations of estradiol on the day of AI. Pregnancies per AI (P/AI) were determined 30 and 65 d after AI. Presynchronization did not affect the concentration of estradiol at AI (DO=6.4 vs. Presynch=5.8 pg/mL), detection of estrus at AI (20.8 vs. 25.9%), or P/AI on d 30 (56.8 vs. 59.1%) and 65 (52.5 vs. 52.4%) after the first AI. Cows receiving COS72 had increased concentration of estradiol (6.6 vs. 5.5 pg/mL) and detection of estrus at AI (28.5 vs. 10.8%) compared with cows receiving COS58. Length of proestrus did not affect P/AI on d 30 (COS72=58.7 vs. COS58=56.1%) but, in Presynch cows, COS58 was detrimental to fertility on d 65 after AI (54.9 vs. 46.5%). Pregnancy loss between gestational d 30 and 65 was greater for Presynch than for DO (7.6 vs. 11.3%), but it was not affected by length of proestrus. Estrous cyclic cows had greater P/AI than anovular cows on d 30 (61.7 vs. 35.1%) and 65 (56.1 vs. 30.7%), but no interaction between estrous cyclic status and treatments was detected. Crossbred Holstein/Jersey cows had superior fertility than their purebred counterparts during the breeding season. The Presynch and DO protocols resulted in similar fertility with no overall difference between the presynchronization methods; however, limiting the length of proestrus to 58 h reduced P/AI in the 5-d timed AI protocol when cows had their estrous cycle presynchronized with Presynch but not with DO.     

Silage chop length and hay supplementation on milk yield, chewing activity, and ruminal digestion by dairy cows

The effects of whole-plant corn silage (CS) particle size and long unprocessed grass hay (LH) supplementation on milk yield, chewing activity, and ruminal digestion in dairy cows were evaluated in 2 experiments. In Experiment 1, corn silage harvested at fine (6 mm; FCS) or coarse (23 mm; CCS) theoretical cut length were fed to 22 lactating Holstein cows. Treatments were 2 total mixed rations containing 58% of dry matter (DM) as FCS or CCS. Diet DM intake tended to be higher in cows fed FCS than those fed CCS (23.4 vs. 22.1 kg/d). However, milk yield and composition, body condition score, and plasma metabolite concentrations were not affected by the dietary treatments. In the second experiment, 5 cannulated Holstein cows were used in a 5 × 5 Latin square design to evaluate the effects of the addition of LH to the diets evaluated in Experiment 1 on chewing activity and ruminal digestion. Treatments were 5 total mixed rations: FCS-based diet plus the addition of 0, 5, or 10% LH (DM basis) and CCS-based diet plus 0 or 5% LH. Long hay addition linearly decreased DM intake in cows fed FCS-based diets (25.0 to 21.7 kg/d), but increased DM intake in those fed CCS-based diets (22.7 to 27.1 kg/d). The intake of neutral detergent fiber (NDF) increased with LH addition in CCS-based diets (7.6 vs. 9.4 kg/d). Rumination time increased (16.8 to 21.0 min/kg of DM intake) when LH was added to FCS-based diets, but it decreased when included in CCS-based diets (18.8 vs. 12.9 min/kg of DM intake). Ruminal pH was higher (5.9 vs. 5.7) and lag-time for in situ NDF disappearance was shorter (3.5 vs. 8.7 h) for cows fed CCS compared with cows fed FCS. The rate of NDF disappearance tended to be higher for the CCS-based diet with 5% LH than for the diet with 0% LH (2.0 vs. 4.4%/h), but solids passage rate was not affected by the treatments. These results suggest that addition of LH to FCS-based diets does not affect ruminal environment or digestion, but depressed DM intake. In contrast, addition of LH to CCS-based diets may improve ruminal NDF digestion, increasing DM intake by reducing filling effect and time needed for rumination.     

Starch and stage of maturity of grass silage: site of digestion and intestinal nutrient supply in dairy cows

The interaction between the quality of grass silage and starch supplementation on ruminal digestion was studied in an experiment with a 2 x 2 factorial design using four dairy cows. Treatment factors were grass silage harvested after either 21 or 37 d of regrowth and two concentrations of steam-flaked corn starch (0 or 4 kg/d). Ruminal volume and flow of duodenal digesta were estimated. When forage was harvested at a more mature stage, only minor effects were noted for silage composition and, consequently, ruminal and intestinal digestion. The addition of starch to the diet tended to reduce ruminal digestion of neutral detergent fiber. The reduction in ruminal digestion was not compensated by increased digestion in the large intestine. Starch increased duodenal nonammonia N flow because of an increase in bacterial N flow. The increase in bacterial N was accompanied by a reduction in the escape of feed N from the rumen. Results from this study indicate that the addition of ruminally available starch to diets based on grass silage reduced ruminally degradable neutral detergent fiber and increased the duodenal supply of protein. These effects have to be taken into account to predict production responses to extra starch.     

Stability of sunflower protein hydrolysates in simulated gastric and intestinal fluids and Caco-2 cell extracts

The in vitro stability of bioactive properties in sunflower protein hydrolysates and purified peptides was studied by using simulated gastric fluid, simulated intestinal fluid, and Caco-2 cells extracts. Protein hydrolysates that inhibit the angiotensin converting enzyme and copper chelating peptides produced by hydrolysis with the microbial protease alcalase were partially resistant to incubation with simulated gastric and intestinal fluids. These hydrolysates and others produced by hydrolysis using pepsin plus pancreatin were also partially resistant to incubation with Caco-2 cells extracts. In addition, the ACE inhibitory peptide FVNPQAGS that is generated by extensive hydrolysis using pepsin and pancreatin, was found to be resistant to hydrolysis by Caco-2 cells extracts. These results suggest that stability in the digestive tract should not be a problem for the bioavailability of these bioactive peptides.     

Influence of gastric digestive reaction on subsequent in vitro intestinal digestion of sodium caseinate-stabilized emulsions

In this study, in vitro intestinal lipid digestion and the physicochemical and microstructural changes of sodium caseinate-stabilized emulsions were examined after the emulsions had been digested in a model simulated gastric fluid containing pepsin for different times. The average size, size distribution, microstructure, proteolysis of interfacial proteins and lipolysis of the emulsion droplets were monitored as a function of digestion time. The emulsion droplets underwent extensive droplet flocculation, with some coalescence together with proteolysis of interfacial proteins, in simulated gastric fluid, resulting in changes in the droplet size and the microstructure of the emulsions. In general, digestion in simulated gastric fluid containing pepsin accelerated coalescence of the emulsion droplets during subsequent digestion in simulated intestinal fluid containing pancreatic lipase. However, the changes in the size, the microstructure and the proteolysis of the interfacial proteins of the emulsions under gastric conditions did not influence the rate and the extent of lipid digestion in the subsequent intestinal environment.     

蚕蛹蛋白和蚕蛹短肽的模拟胃肠道消化特性研究

研究了蚕蛹蛋白和蚕蛹短肽的氨基酸组成,并分析了其在模拟胃肠道消化过程中的消化特性和氮释放量,探究蚕蛹蛋白和短肽在人类营养支持方面的应用。使用自动氨基酸分析仪测定蚕蛹蛋白和蚕蛹短肽的氨基酸含量,并计算氨基酸评分(AAS)、化学评分(CS)和必需氨基酸指数(EAAI)。蚕蛹蛋白和蚕蛹短肽的消化特性和氮释放量通过体外模拟胃肠道消化实验和凯氏定氮法进行评价的计算。结果表明,蚕蛹蛋白和蚕蛹短肽含有人体所需的18种氨基酸,而且必需氨基酸的组成合理。蚕蛹蛋白和蚕蛹短肽在体外模拟胃肠道消化实验中有良好高效的消化率和较高的氮释放量(约90%)。因此在今后将蚕蛹蛋白和蚕蛹短肽作为优质且经济的蛋白质原料,在动物饲料和人类肠内营养支持制剂方面有巨大的应用潜力。