Effects Of Direct-fed Microbial Supplementation Research Articles

Effects of direct-fed microbial supplementation on performance and immune parameters of lactating dairy cows

We evaluated the effects of supplementing bacterial direct-fed microbial (DFM) on performance, apparent total-tract digestibility, rumen fermentation, and immune parameters of lactating dairy cows. One hundred fourteen multiparous Holstein cows (41 ± 7 DIM) were used in a randomized complete block design with an experiment comprising 14 d of a covariate (pre-experimental sample and data collection) and 91 d of an experimental period. Cows were blocked based on energy-corrected milk (ECM) yield during the covariate period and the following treatments were randomly assigned within each block: (1) control (CON), corn silage-based total mixed ration without DFM; (2) PRO-A, basal diet top-dressed with a mixture of Lactobacillus animalis and Propionibacterium freudenreichii at 3 × 109 cfu/d; and 3) PRO-B, basal diet top-dressed with a mixture of L. animalis, P. freudenreichii, Bacillus subtilis, and Bacillus licheniformis at 11.8 × 109 cfu/d. Milk yield, dry matter intake (DMI), and body weight were measured daily, while milk samples for component analysis were taken on 2 consecutive days of each week of data collection. Feces, urine, rumen, and blood samples were taken during the covariate period, wk 4, 7, 10, and 13 for estimation of digestibility, N-partitioning, rumen fermentation, plasma nutrient status and immune parameters. Treatments had no effect on DMI and milk yield. Fat-corrected milk (3.5% FCM) and milk fat yield were improved with PRO-B, while milk fat percent and feed efficiency (ECM/DMI) tended to increase with PRO-B compared with PRO-A and CON. Crude fat digestibility was greater with PRO-B compared with CON. Feeding CON and PRO-A resulted in higher total volatile fatty acid concentration relative to PRO-B. Percentage of neutrophils tended to be reduced with PRO-A compared with CON and PRO-B. The mean fluorescence intensity (MFI) of anti-CD44 antibody on granulocytes tended to be higher in PRO-B compared with CON. The MFI of anti-CD62L antibody on CD8+ T cells was lower in PRO-A than PRO-B, with PRO-A also showing a tendency to be lower than CON. This study indicates the potential of DFM to improve fat digestibility with consequential improvement in fat corrected milk yield, feed efficiency and milk fat yield by lactating dairy cows. The study findings also indicate that dietary supplementation with DFM may augment immune parameters or activation of immune cells, including granulocytes and T cells; however, the overall effects on immune parameters are inconclusive.

Evaluating the effects of direct-fed microbial supplementation on the performance, milk quality and fatty acid of mid-lactating dairy cows.

The objective of the experiment was to investigate the effect of a mixture of direct-fed microbial (DFM) on feed intake, nutrient digestibility, milk yield and composition, milk fatty acid and blood parameter in crossbred mid-lactating cows. Twenty-four crossbred Holstein cows (body weight = 650±15kg; days in milk = 100±20; daily milk yield = 25±3kg) were used in a completely randomized design with three treatments: (1) CON, without DFM; (2) LS, inoculation with Lactobacillus fermentum (4.5 × 108 CFU/day) plus Saccharomyces cerevisiae (1.4×1010 CFU/day); and (3) LSM, inoculation with LS plus Megasphaera elsdenii (4.5 × 108 CFU/day). All animals received the same ration with 45.7% forage and 54.3% concentrate. Results showed that the highest feed intake was observed in treatments LS and LSM (p = 0.02). Compared with the CON, milk production, 4% fat-corrected milk, energy-corrected milk, fat (kg/day), protein (kg/day) and lactose (kg/day), FE and percent of fat were increased (p<0.05) by LSM, but unaffected by LS. Also, compared with the CON, both LS and LSM increased antioxidant activity (p<0.05). The concentration of C18:2c n-6 increased significantly in treatment LSM compared with the CON (p = 0.003). The concentration of C20:0 increased significantly in treatment LS compared with the CON (p = 0.004). The highest concentrations of insulin, glucose, triglyceride and cholesterol were observed by LSM (p<0.05). Compared with the CON, both LS and LSM increased blood monocyte, neutrophil, eosinophil and basophil (p<0.05), and blood lymphocyte was increased (p = 0.02) only by LSM. The results of the research showed that the use of DFMs had no effect on the digestibility, microbial load and the major part of fatty acids in milk. However, it improved feed intake, milk yield and antioxidant activity of milk and also increased the milk concentration of C18:2 n-6.

Carryover effect of direct-fed microbial supplementation and early weaning on the growth performance and carcass characteristics of growing Najdi lambs

Abstract Growing Najdi lambs were randomly selected from lambs weaned at 30, 45, and 60 days old which were treated with 3 doses of direct-fed microbial (DFM) at 5, 10, and 15 days old to investigate the carryover effect of early weaning and DFM supplementation on their growth performance and carcass characteristics. Ten lambs from each group were transferred to individual pens for a feeding trial using the total mixed ration (Wafi). Lambs treated with DFM and weaned at 60 days old showed numerically higher body weight and average daily gains compared to other groups. Concerning the carcass and meat quality, there were significant differences between all groups in all carcass and tissue measurements, except for the back-fat and body wall thickness. In conclusion, treatment did not have any significant negative effect on body weight, feed intake, and conversion ratio compared with the control, but positively affected Zn and Cu absorption. DFM also played an important role in fat metabolism, which affects fat deposits in carcasses. The most important finding was that early weaning can be performed using DFM supplementation without any negative effect on the lambs’ performance during growth.

Effects of direct-fed microbial supplementation on growth performance and metabolic profile of newborn lambs weaned at different ages

The aim of this study was to investigate the effects of direct fed microbial (DFM) supplementation on growth performance, metabolic profile and mortality rate of newborn lambs at different weaning ages. Five days old sixty Najdi male lambs (5.40 ± 0.10 kg BW) were randomly assigned to 1 of 4 treatments (15 lambs each): lambs received either no DFM and weaned at 60 days of age (control) or oral dose of DFM (5 ml) at 5, 10 and 15 days old, and weaned at 30 (DFM30), 45 (DFM45) or 60 (DFM60) days old, respectively. Final body weight was higher (P=0.05) and tended to be higher (P=0.09) in DFM30 and DFM45 lambs, respectively whereas body weight gain was higher (P=0.01) in DFM45 and DFM60 lambs. The mortality rate was declined from 33.3s% to 6.67% in DFM30 and DFM45 lambs compared to control ones. Serum cortisol and creatinine levels tended to be reduced (P=0.07 and 0.11, respectively) in DFM30 and DFM45 lambs. The serum total cholesterol was highest (P=0.02) in DFM60 lambs and lowest in both DFM45 and control lambs. Similarly, the DFM30 and DFM45 lambs showed a declined serum triglyceride compared with control and DFM60 lambs (P=0.02). Serum phosphorus declined (P=0.04) in DFM30 treated lambs while serum zinc and copper levels decreased (P=0.02 and 0.05, respectively) in all treatments compared to control lambs. These results indicated that DFM supplementation may enhance the growth and health of early weaned lambs.

Effect of direct-fed microbials on culturable gut microbiotas in broiler chickens: a meta-analysis of controlled trials.

ObjectiveThis meta-analysis was conducted to evaluate the overall effect of direct-fed microbial (DFM) or probiotic supplementation on the log concentrations of culturable gut microbiota in broiler chickens.MethodsRelevant studies were collected from PubMed, SCOPUS, Poultry Science Journal, and Google Scholar. The studies included controlled trials using DFM supplementation in broiler chickens and reporting log concentrations of the culturable gut microbiota. The overall effect of DFM supplementation was determined using standardized mean difference (SMD) with a random-effects model. Subgroups were analyzed to identify pre-specified characteristics possibly associated with the heterogeneity of the results. Risk of bias and publication bias were assessed.ResultsEighteen taxa of the culturable gut microbiota were identified from 42 studies. The overall effect of DFM supplementation on the log concentrations of all 18 taxa did not differ significantly from the controls (SMD = −0.06, 95% confidence interval [−0.16, 0.04], p = 0.228, I2 = 85%, n = 699 comparisons), but the 18 taxa could be further classified into three categories by the direction of the effect size: taxa whose log concentrations did not differ significantly from the controls (category 1), taxa whose log concentrations increased significantly with DFM supplementation (category 2), and taxa whose log concentrations decreased significantly with DFM supplementation (category 3). Category 1 comprised nine taxa, including total bacterial counts. Category 2 comprised four taxa: Bacillus, Bifidobacterium, Clostridium butyricum, and Lactobacillus. Category 3 comprised five taxa: Clostridium perfringens, coliforms, Escherichia coli, Enterococcus, and Salmonella. Some characteristics identified by the subgroup analysis were associated with result heterogeneity. Most studies, however, were present with unclear risk of bias. Publication bias was also identified.ConclusionDFM supplementation increased the concentrations of some beneficial bacteria (e.g. Bifidobacterium and Lactobacillus) and decreased those of some detrimental bacteria (e.g. Clostridium perfringens and Salmonella) in the guts of broiler chickens.

The effects of direct-fed microbial supplementation, as an alternative to antibiotics, on growth performance, intestinal immune status and epithelial barrier protein expression in broiler chickens

Abstract This study was conducted to investigate the effects of Bacillus subtilis supplementation in broiler chicken diets on growth performance, feed efficiency, intestinal cytokine and tight junction (TJ) protein mRNA expression. Day-old broiler chicks (n = 140) were to one of five dietary treatments: basal diet (CON), basal diet supplemented with antibiotic (BMD) or probiotics, namely B. subtilis strain 1781 (PB1), a combination of B. subtilis strain 1104+strain 747 (PB2) or B. subtilis strain 1781+strain 747 (PB3). At 14 days of age, ileal samples were collected and used for intestinal cytokine, TJ protein and mucin gene expression analysis using qRT-PCR. The chickens fed with antibiotic (BMD) and B. subtilis strain 1781 alone (PB1) had significantly higher body weights compared to controls of the same age. Dietary supplementation with antibiotic (BMD) or probiotics (PB1, PB2, PB3) significantly improved the feed efficiency as evidenced by decreased FCR compared to controls. No differences were observed in the expression of IL1β, IL17F, IFNγ and MUC2 gene among the different treatment groups. However, elevated expression of IL6 (BMD, PB1, PB2), IL8 (PB2) and TNFSF15 (PB1, PB2, PB3) compared to controls was observed in the ileum. IL2 and IL10 expression was upregulated in chicks in the PB2 and PB3 groups, and IL4 was elevated in the PB1 group. IL13 was elevated in all probiotic-fed groups. Probiotic supplementation was also shown to significantly increase the expression of TJ proteins JAM2, ZO1 (PB2, PB3) and occludin (PB1, PB2). Taken together, B. subtilis supplementation altered intestinal immune activity and influenced gut barrier integrity through increased expression of TJ proteins.

The Effects of Direct-fed Microbial Supplementation, as an Alternative to Antibiotics, on Growth Performance, Intestinal Immune Status, and Epithelial Barrier Gene Expression in Broiler Chickens

The objective of this study was to investigate the effects of Bacillus subtilis-based probiotic supplementation in broiler chicken diets on growth performance, feed efficiency, intestinal cytokine, and tight junction (TJ) protein mRNA expression. Zero-day-old broiler chicks (n=140) were randomly assigned to one of five dietary treatments: basal diet (CON); basal diet supplemented with either antibiotic bacitracin methylene disalicylate (BMD); or probiotics, namely, B. subtilis strain 1781 (PB1), a combination of B. subtilis strain 1104+strain 747 (PB2), or B. subtilis strain 1781+strain 747 (PB3). Body weight and feed intake were measured at 14days of age, and the feed conversion ratio (FCR) was calculated. At 14days of age, ileal samples were collected and used for intestinal cytokine, TJ protein, and mucin gene expression analysis using qRT-PCR. The chickens supplemented with antibiotic (BMD) and B. subtilis strain 1781 alone (PB1) had significantly higher body weights compared to controls of the same age. Dietary supplementation with antibiotic (BMD) or probiotics (PB1, PB2, PB3) significantly improved the feed efficiency as evidenced by decreased FCR compared to controls. No differences were observed in the expression of IL1β, IL17F, IFNγ, and MUC2 gene among the different treatment groups. However, elevated expression of IL6 (BMD, PB1, PB2), IL8 (PB2), and TNFSF15 (PB1, PB2, PB3) compared to controls was observed in the ileum. IL2 and IL10 expression was upregulated in chicks in the PB2 and PB3 groups, and IL4 was elevated in the PB1 group. IL13 was elevated in all probiotic-fed groups (PB1, PB2, PB3). Probiotic supplementation was also shown to significantly increase the expression of TJ proteins JAM2, ZO1 (PB2, PB3), and occludin (PB1, PB2). Taken together, B. subtilis supplementation altered intestinal immune activity and influenced gut barrier integrity through increased tight junction gene expression.

Effect of direct-fed microbial supplementation on lactation performance and total-tract starch digestibility by midlactation dairy cows

ABSTRACT The objective of this study was to evaluate the effect of direct-fed-microbial addition to a TMR on lactation performance by midlactation dairy cows. A total of 112 Holstein cows (28 primiparous and 84 multiparous; 139 ± 47 DIM at trial initiation) were stratified by parity and DIM and randomly assigned to 14 pens of 8 cows each. Pens were then randomly assigned to 1 of 2 treatments in a continuous-lactation trial consisting of a 2-wk covariate adjustment period with cows fed the basal TMR followed by a 10-wk treatment period with cows fed their assigned treatment diets. The 2 treatments were basal TMR plus either 1 g/cow per day of a direct-fed microbial (BOV, 1 × 10 9 cfu/g Lactobacillus acidophilus NP51 and 2 × 10 9 cfu/g Propionibacterium freudenreichii NP24) or placebo (control). Milk yield was similar between treatments and averaged 44.9 kg/d. There was a trend (P 0.10) by treatment. Milk fat, protein and urea nitrogen concentrations were unaffected (P > 0.10) by treatment and averaged 3.75%, 3.11%, and 15.2 mg/dL, respectively. Likewise, BW change and condition score did not differ (P > 0.10). Under the conditions of this study, with midlactation cows fed a 54% forage TMR (DM basis), there was no improvement in lactation performance from the inclusion of a direct-fed microbial in the diet.

Effects of direct-fed microbial supplementation on broiler performance, intestinal nutrient transport and integrity under experimental conditions with increased microbial challenge

1. The effects of Aspergillus oryzae- and Bacillus subtilis-based direct-fed microbials (DFM) were investigated on the performance, ileal nutrient transport and intestinal integrity of broiler chickens, raised under experimental conditions, with increased intestinal microbial challenge.2. The first study was a 3 × 2 factorial experiment, with 3 dietary treatments (control (CON), CON + DFM and CON + antibiotic growth promoter) with and without challenge. Chicks were fed experimental diets from 1 to 28 d, while the challenge was provided by vaccinating with 10 times the normal dose of commercial coccidial vaccine on d 9. In a second experiment, two groups of 1 d-old broilers, housed on built-up litter (uncleaned from two previous flocks), were fed the same CON and CON + DFM diets from 1 to 21 d.3. The challenge in the first experiment reduced performance, but no differences were observed among dietary treatments from 8 to 28 d. The challenge reduced the ileal epithelial flux for D-glucose, L-lysine, DL-methionine and phosphorus on d 21. Epithelial flux for D-glucose, L-lysine and DL-methionine were increased by DFM. Ileal trans-epithelial electrical resistance (TER) was increased in challenged broilers fed DFM, although this was not observed in unchallenged birds as indicated by a significant interaction.4. Ileal mucin mRNA expression and colon TER were increased, and colon endotoxin permeability was reduced by DFM on d 21 in the second experiment.5. It was concluded that the addition of DFM in the diet improved the intestinal integrity of broiler chickens raised under experimental conditions designed to provide increased intestinal microbial challenge.

Investigation on the effects of adding Direct– fed Microbial on total tract apparent nutrient digestibility in sheep

Growing concern over the use of antibiotics and other growth stimulation in the animal feed industry has increased interest in evaluating the effects of Direct-fed Microbial (DFM) on animal performance (Sullivan and Martin, 1999). Some bacterial direct-fed microbials also seem able to function in the rumen. Depending on the species or combination of species, they can increase digestibility. Modes of action are variable and include native rumen microbiota enhancement through increased substrate breakdown, enhancement of nutrient uptake, stabilising rumen pH and provision of growth factors. The objectives of this study were to evaluate the effects of DFM supplementation on total tract digestibility.

Effects of direct-fed microbial supplementation on digestibility and fermentation end-products in horses fed low- and high-starch concentrates1

A study was conducted to determine whether direct-fed microbials (DFM) could be used to increase digestibility and minimize the risk of acidosis associated with feeding an increase in the amount of starch fed to horses. Fifteen mature Thoroughbred geldings were randomly assigned to 1 of 3 treatments in a 3 x 3 Latin square design balanced for carryover effects. Within each 26-d period, horses were offered grass hay + low-starch concentrate (LS; 1.2 g of starch x kg of BW(-1) x meal(-1)) from d 1 to 13 and then were abruptly changed to hay + high-starch concentrate (HS; 2.4 g of starch.kg of BW(-1)x meal(-1)) on d 14 continuing through d 26. The DFM treatments were offered in concentrate pellets at a target dosage of 10(8) cfu/(50 kg of BW x d) as follows: no DFM (CON; control), Lactobacillus acidophilus (LAC1; single-species DFM), or a mixture of L. acidophilus, Lactobacillus casei, Bifidobacterium bifidum, and Enterococcus faecium (LAC4; multiple-species DFM). Total feces were collected over 72 h from d 11 to 13 (LS; low dietary starch collection), from d 15 to 17 (AC; abrupt change in dietary starch collection), and at the end of each experimental period, from d 24 to 26 (HS; high dietary starch collection). Data collected consisted of total DM intake and fecal output, fecal pH, fecal acetate and propionate concentrations, and viable numbers of DFM in the feed. With the exception of Fe digestibility, there were no starch x DFM interactions. There was an effect of starch level (P <or= 0.02) on most nutrient digestibilities, except for Mg (P = 0.08) and CP, K, and Zn (P > 0.10). Horses supplemented with LAC4 had increased ether extract (P < 0.05) and a tendency for decreased Na (P < 0.10) digestibilities compared with CON horses. All DFM-supplemented horses had increased Cu (P < 0.05) and Fe and numerically increased Zn digestibilities compared with CON horses. Fecal pH decreased (P < 0.05), and fecal propionate concentration increased (P < 0.05) as dietary starch content changed from LS to HS. There was a tendency for elevated fecal pH (P < 0.10) in LAC1 horses compared with CON horses. These results confirm that increasing starch in the equine diet can enhance nutrient digestibility of the diet. Supplementing equine diets with either a single or mixed strain direct-fed lactic acid bacteria had limited effects on nutrient digestibility or on reducing the risk of acidosis associated with feeding high-starch concentrates to horses. The potential response of DFM supplementation should be evaluated when a more acute acidotic state is induced in horses than in the current study.

Effects of Lactobacillus reuteri-based Direct-fed Microbial Supplementation for Growing-Finishing Pigs

Two experiments were conducted to evaluate the effects of direct-fed microbial supplementation on the growth performance and nutrient digestibility in pigs. In experiment 1, forty eight pigs were used in a 42-d growth assay. There were four pigs per pen and three pens per treatment. Dietary treatments included 1) NC (without antibiotic basal diet), 2) PC (NC diet+0.1% antibiotic, 100 g/kg chlortetracycline), 3) DFM-1 (NC diet+0.2% Lactobacillus reuteri and Lactobacillus salivarius complex) and 4) DFM-2 (NC diet+0.2% Lactobacillus reuteri, Lactobacillus salivarius, Lactobacillus plantarum and Yeast complex). For the overall period, no treatment had significant effects in growth performance. In the nutrition digestibility, the pigs fed DFM diets were improved in DM and N digestibility compared with the pigs fed NC and PC diets but it was not significantly different. In experiment 2, sixty four crossbred pigs were used in a 98-d growth assay. There were four pigs per pen and four pens per treatment. Dietary treatments included 1) HND (high nutrient diet), 2) LND (low nutrient diet), 3) HND+DFM (HND diet+0.2% Lactobacillus reuteri and Lactobacillus plantarum complex) and 4) LND+DFM (LND diet+0.2% Lactobacillus reuteri and Lactobacillus plantarum complex). For overall period of growing phase, the pigs fed LND diets had improved gain/feed (p=0.01) and for overall period in the finishing phase, the pigs fed LND with DFM diets had higher ADG, ADFI and gain/feed than the others but there were no significant differences (p>0.13). In total period of growing-finishing phase, the pigs fed LND diet had higher gain/feed than the pigs fed HND diets (p<0.05). In growing phase, there were not significant differences among the treatments means for DM and N digestibility. However, the pigs fed diets with DFM had improved N digestibility (p<0.02) compared to the pigs fed diets without DFM in finishing phase. In conclusion, DFM slightly improved the growth performance in growing-finishing pigs. (Asian-Aust. J. Anim. Sci. 2005. Vol 18, No. 3 : 370-374)

Effects of direct-fed microbial supplementation on ruminal acidosis, digestibility, and bacterial protein synthesis in continuous culture

A study was conducted to determine whether a bacterial direct-fed microbial (DFM) alone or with yeast could be used to minimize the risk of ruminal acidosis and to improve utilization of a feedlot cattle diet containing high concentrate. A dual effluent continuous culture (CC) system was used to investigate the effects of DFM on fermentation, digestibility, and microbial protein synthesis in a 4×4 Latin square design. Treatments were control (without DFM), Propionibacterium P15 (PB), Enterococcus faecium EF212 (EF), and E. faecium EF212 combined with a yeast, Saccharomyces cerevisiae (EFY) (Chr. Hansen Inc., Milwaukee, WI). Fermenters were fed twice daily a feedlot finishing cattle diet that consisted of about 871 g/kg barley grain, 79 g/kg barley silage and 50 g/kg supplement (dry matter (DM) basis). The DFM products (1×10 9 or 6×10 8 colony-forming units (CFU)/g for PB or EF and EFY, respectively) were delivered equally twice daily into the fermenters just before feed provision. Mean fermenter pH ranged from 5.86 to 5.91, and did not differ among the treatments. Total VFA concentration and its molar proportions were not affected by the DFM supplementation except for caproic acid which was higher ( P<0.05) for control than for DFM addition. The ratio of acetate to propionate was similar among the treatments but was relatively lower (range of 0.83–0.93) than that usually observed in the rumen of feedlot cattle (range of 1.5–2.0) due to a considerably lower proportion of acetate (395 mmol/mol) but high proportion of propionate (460 mmol/mol). Ruminal lactate-utilizing bacterial numbers were ( P<0.05) greater for control and EF diets than for PB or EFY diets. In vitro digestibilities of DM, organic matter, starch and crude protein were all in the range of in vivo findings but digestibilities of fiber including acid or neutral detergent fibers were significantly lower than those of in vivo findings. However, no differences in digestion of any nutrients studied were detected among the treatments. Microbial protein synthesis (range of 0.9–1.0 g nitrogen [N]/day) and microbial efficiency (range of 18–20 g N/kg of truly fermented organic matter) were also similar among the treatments. The present results indicate that addition of DFM such as PB, EF or EF combined with yeast had no effect on preventing ruminal acidosis and on fermentation or nutrient digestion in CC. The CC technique can be used as an alternative to in vivo techniques to assess effect of DFM supplementation on ruminal acidosis and digestion of nutrients.