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Probiotics are live microorganisms which, when administered in adequate amounts, confer health benefits to the host. The use of probiotics in poultry has increased steadily over the years due to higher demand for antibiotic-free poultry. The objective of this systematic review is to present and evaluate the effects of probiotics on the nutrient utilization, growth and laying performance, gut histomorphology, immunity, and gut microbiota of poultry. An electronic search was conducted using relevant keywords to include papers pertinent to the topic. Seventeen commonly used probiotic species were critically assessed for their roles in the performance and gut health of poultry under existing commercial production conditions. The results showed that probiotic supplementation could have the following effects: (1) modification of the intestinal microbiota, (2) stimulation of the immune system, (3) reduction in inflammatory reactions, (4) prevention of pathogen colonization, (5) enhancement of growth performance, (6) alteration of the ileal digestibility and total tract apparent digestibility coefficient, and (7) decrease in ammonia and urea excretion. Thus, probiotics can serve as a potential alternative to antibiotic growth promoters in poultry production. However, factors such as the intestinal health condition of birds, the probiotic inclusion level; and the incubation conditions, feedstuff, and water quality offered to birds may affect the outcome. This systematic review provides a summary of the use of probiotics in poultry production, as well as the potential role of probiotics in the nutrient utilization, growth and laying performance, and gut health of poultry.

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±15 kg; days in milk = 100±20; daily milk yield = 25±3 kg) were used in a completely randomized design with three treatments: (1) CON, without DFM; (2) LS, inoculation with Lactobacillus fermentum (4.5 × 10 CFU/day) plus Saccharomyces cerevisiae (1.4×10 CFU/day); and (3) LSM, inoculation with LS plus Megasphaera elsdenii (4.5 × 10 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.

Direct-fed microbials (DFM) could serve as a potential alternative to the feeding of antibiotics in poultry production. In this study, the effects of providing a DFM were compared with the feeding of salinomycin on intestinal histomorphometrics, and microarchitecture was examined. Broiler chicks (n=18 per treatment; trials 1 and 2) were fed a standard starter diet (control), control+PrimaLac (DFM; 0.3% wt/wt), and control+salinomycin (SAL; 50 ppm) from hatch to 21d. The birds were euthanized on d 21, and the ileal, jejunal, cecal, and colon tissues were dissected. Samples were examined by light microscopy (jejunum and ileum; trial 1) and scanning electron microscopy (ileum, cecum, and colon; trial 2). Feeding of the DFM increased intestinal muscle thickness (P<0.05) up to 33% compared with the control treatment. The DFM group also had increased villus height and perimeter (P=0.009 and 0.003, respectively) in jejunum. Segmented filamentous-like bacteria were less numerous in DFM-treated chicks than in the control chicks. Very few segmented filamentous-like bacteria were found near other microbes in the ileum. The DFM chicks had a larger number of bacteria positioned over or near goblet cells and in intervilli spaces. Bacteria in the colon were observed to be attached primarily around and within the crypts. Mucous thickness was less, and the density of bacteria embedded in the mucous blanket appeared to be lower in DFM-treated animals than in the control in all intestinal segments. The birds fed SAL had fewer bacteria and enterocytes in the ileum than in the control-and DFM-treated birds, and they had thicker and fewer microvilli. Because gastrointestinal track colonization by the DFM organisms can prevent the attachment of pathogens to the epithelium, spatial relationships, in this study, demonstrate the functionality of DFM and probiotics in preventing disease. It also supports previous observations that the feeding of salinomycin may alter intestinal function.

The current study investigated whole-body O2 consumption, intestinal O2 consumption, and intestinal inflammation status through mucosal cytokine production on broiler chicks fed the direct-fed microbial PrimaLac. One hundred twenty 1-d-old broiler chicks were randomly assigned to 1 of 3 experimental diets: standard starter diet (control), standard starter diet with added salinomycin (SAL), and standard starter diet with added PrimaLac (DFM). Birds were housed in 2 separate rooms, the control and SAL treatments in one room and the DFM in another. Intact ileal and cecal samples were collected on d 19, 20, and 21 after measuring whole-body O2 consumption using indirect calorimetry. The O2 up-take of ileal tissue was measured using an in vitro O2 monitor. Analysis of intestinal immune status of broilers was measured by the relative differences in mRNA of both pro- and antiinflammatory cytokines: interleukin-(IL) 1beta, IL-6, and IL-10 using real-time reverse transcription-PCR. Broilers exhibited a 6 to 16% decrease in whole-body energy expenditures and up to a 47% decrease (P<0.05) in ileal energy expenditures in the DFM group compared with other treatments. The reverse transcription-PCR data demonstrated that DFM consortium numerically altered both pro- and antiinflammatory cytokines within the ileum of 19-d posthatch broilers. These data suggest that direct-fed microbials like PrimaLac increase metabolic efficiency via changes in intestinal physiology and metabolism.

Abstract The objective of this experiment was to investigate the impact of an F18 enterotoxigenic Escherichia coli (ETEC) challenge on growth performance, aspects of intestinal function, and selected immune responses of piglets, as well as to evaluate potential protective effects of direct-fed microbial (DFM) blends. Seventy-two weaned piglets (6.4 ± 0.2 kg body weight [BW]; ~21 d of age) were assigned to one of four treatments: 1) NC: Nonchallenged (n = 10), 2) positive challenged control (PC): F18 ETEC-challenged (n = 10), 3) PC + DFM1 (n = 8; three strains of Bacillus amyloliquefaciens; 7.5 × 105 colony-forming units [cfu]/g), or 4) PC + DFM2 (n=8; 2 strains of B. amyloliquefaciens and one strain of Bacillus subtilis; 1.5 × 105 cfu/g). Feed intake and BW were recorded on day 0, 7, and 17. Pigs were sham-infected either with 6 mL phosphate-buffered saline or inoculated with 6 mL F18 ETEC (~1.9 × 109 cfu/mL) on day 7 (0 d postinoculation [dpi]). All ETEC-challenged pigs were confirmed to be genetically susceptible to F18. Pigs had ad libitum access to feed and water throughout the 17-d trial. Fecal scores were visually ranked and rectal temperatures were recorded daily. To evaluate ETEC shedding, fecal swabs were collected on dpi 0, 1, 2, 3, 5, 7, and 10. Blood samples were collected on dpi 0, 1, 2, 4, 7, and 10. Ileal tissues were collected at necropsy on dpi 10. All challenged treatments had lower final BW, decreased average daily gain (ADG), and average daily feed intake (ADFI) during the 10-d postchallenge period (P < 0.01). The DFM2 treatment increased E. coli shedding on dpi 2 and decreased iton dpi 7 (P < 0.05) compared with the PC. Rectal temperature decreased across all challenged treatments (P < 0.01). Ileal mRNA abundance of occludin (OCLN) and zonula occludens-1 (ZO-1) decreased in PC and DFM1 compared with NC (P < 0.05). Pigs fed DFM2 had intermediate ileal mRNA abundance of OCLN and increased ZO-1 mRNA compared with pigs in PC (P < 0.05). Interleukin 8 (IL-8) increased in the plasma of PC and DFM2 on dpi 2 compared with NC (P < 0.05). Mucosal IL-8 increased in PC compared with NC (P < 0.05). All challenged treatments tended to have elevated tumor necrosis factor-α (TNF-α) mRNA abundance compared with NC (P < 0.10). Challenged pigs had reduced secretory immunoglobulin A and villus height compared with NC pigs (P < 0.05). The impact of an ETEC challenge on intestinal function and the immune system has been revealed, information critical to developing improved treatment regimes.

Two experiments were conducted to investigate the effects of oral supplementation of the lactic-acid-producing bacterium Enterococcus faecium EF212 alone or in combination with Saccharomyces cerevisiae (yeast) on mediators of the acute phase response in feedlot steers. Eight fistulated steers were used to study the effects of E. faecium alone or with yeast in a crossover design with 2 Latin squares, 4 steers within each square, and 2 periods. The length of each period was 3 wk, with a 10-d adaptation and an 11-d measurement period. The experimental diet contained 87% steam-rolled barley, 8% whole-crop barley silage, and 5% supplement (DM basis). In Exp. 1, treatments were control vs. the lactic-acid-producing bacterium E. faecium (6 x 10¹⁰ cfu/d). In Exp. 2, treatments were control vs. E. faecium (6 x 10¹⁰ cfu/d) and S. cerevisiae (6 x 10¹⁰ cfu/d). The bacteria and yeast supplements were blended with calcium carbonate to supply 6 x 10¹⁰ cfu/d when top-dressed into the diet once daily at the time of feeding (10 g/d). Steers fed the control diet received only carrier (10 g/d). Blood samples were collected from the jugular vein on d 17 and 21 of each period, and serum amyloid A (SAA), lipopolysaccharide binding protein (LBP), haptoglobin, and alpha₁-acid glycoprotein (α₁-AGP) were measured. Supplementation of feed with E. faecium had no effect on concentrations of SAA, LBP, haptoglobin, or α₁-AGP in plasma compared with those of controls. However, feeding E. faecium and yeast increased (P = 0.02) plasma concentrations of SAA, LBP, and haptoglobin but had no effect on plasma α₁-AGP. In conclusion, oral supplementation of E. faecium alone had no effect on the mediators of the acute phase response that were measured, whereas feeding of E. faecium and yeast induced an inflammatory response in feedlot steers fed high-grain diets. Further research is warranted to determine the mechanism(s) by which E. faecium and yeast stimulated production of acute phase proteins in feedlot steers.

We examined the effects of two direct-fed microbials (DFM) containing multiple microbial species and their fermentation products on energy status, nutrient digestibility, and ruminal fermentation, bacterial community, and metabolome of beef steers. Nine ruminally cannulated Holstein steers (mean ± SD body weight: 243 ± 12.4 kg) were assigned to three treatments arranged in a triplicated 3 x 3 Latin square design with three 21-d periods. Dietary treatments were 1) control (CON; basal diet), 2) Commence (PROB; basal diet plus 19 g/d of Commence), and 3) RX3 (SYNB; basal diet plus 28 g/d of RX3). Commence and RX3 are both multispecies DFM products. From day 16 to 20 of each period, feed and fecal samples were collected daily to determine the apparent total tract digestibilities of nutrients using indigestible neutral detergent fiber method. On day 21 of each period, blood samples were collected for analysis of plasma glucose and nonesterified fatty acid. Ruminal contents were collected at approximately 1, 3, 6, 9, 12, and 18 h after feeding on day 21 for analysis of volatile fatty acids (VFA), lactate, ammonia-N concentrations, bacterial community, and metabolome profile. Total tract digestibilities of nutrients did not differ (P > 0.05) among treatments. Compared with CON, steers fed either supplemental PROB or SYNB had greater (P = 0.04) plasma glucose concentrations. Compared with CON, total ruminal VFA, propionate, isovalerate, and valerate concentrations increased (P < 0.05) or tended to increase (P < 0.10) with either supplemental PROB or SYNB, but were not different (P > 0.05) between PROB and SYNB. Compared with CON, PROB reduced (P < 0.05) the relative abundance of Prevotella 1 and Prevotellaceae UCG-001 but increased (P < 0.05) the relative abundance of Rikenellaceae RC9, Succinivibrionaceae UCG-001, Succiniclasticum, and Ruminococcaceae UCG-002. Supplemental SYNB decreased (P < 0.05) the relative abundance of Prevotella 1 and Prevotellaceae UCG-001 but increased (P < 0.05) the relative abundance of Prevotella 7, Succinivibrio, Succiniclasticum, and Ruminococcaceae UCG-014. Compared with CON, metabolome analysis revealed that some amino acids were increased (P < 0.05) in steers fed PROB. This study demonstrated that, compared with CON, supplementation of either PROB or SYNB altered the ruminal bacterial community and metabolome differently; however, their effects on the ruminal VFA profile and energy status of the steers were not different from each other.

Social concern about misuse of antibiotics as growth promoters (AGP) and generation of multidrug-resistant bacteria have restricted the dietary inclusion of antibiotics in livestock feed in several countries. Direct-fed microbials (DFM) are one of the multiple alternatives commonly evaluated as substitutes of AGP. Sporeformer bacteria from the genus have been extensively investigated because of their extraordinary properties to form highly resistant endospores, produce antimicrobial compounds, and synthesize different exogenous enzymes. The purpose of the present study was to evaluate and select spp. from environmental and poultry sources as DFM candidates, considering their enzyme production profile, biofilm synthesis capacity, and pathogen-inhibition activity. Thirty-one isolates were screened for relative enzyme activity of amylase, protease, lipase, and phytase using a selective media for each enzyme, with 3/31 strains selected as superior enzyme producers. These three isolates were identified as (1/3), and (2/3), based on biochemical tests and 16S rRNA sequence analysis. For evaluation of biofilm synthesis, the generation of an adherent crystal violet-stained ring was determined in polypropylene tubes, resulting in 11/31 strains showing a strong biofilm formation. Moreover, all strains were evaluated for growth inhibition activity against serovar Enteritidis (26/31), (28/31), and (29/31). Additionally, in previous and studies, these selected strains have shown to be resistant to different biochemical conditions of the gastrointestinal tract of poultry. Results of the present study suggest that the selection and consumption of -DFM, producing a variable set of enzymes and antimicrobial compounds, may contribute to enhanced performance through improving nutrient digestibility, reducing intestinal viscosity, maintaining a beneficial gut microbiota, and promoting healthy intestinal integrity in poultry.

Direct-fed microbials (DFMs) are feed additives containing live naturally existing microbes that can benefit animals’ health and production performance. Due to the banned or strictly limited prophylactic and growth promoting usage of antibiotics, DFMs have been considered as one of antimicrobial alternatives in livestock industry. Microorganisms used as DFMs for ruminants usually consist of bacteria including lactic acid producing bacteria, lactic acid utilizing bacteria and other bacterial groups, and fungi containing Saccharomyces and Aspergillus . To date, the available DFMs for ruminants have been largely based on their effects on improving the feed efficiency and ruminant productivity through enhancing the rumen function such as stabilizing ruminal pH, promoting ruminal fermentation and feed digestion. Recent research has shown emerging evidence that the DFMs may improve performance and health in young ruminants, however, these positive outcomes were not consistent among studies and the modes of action have not been clearly defined. This review summarizes the DFM studies conducted in ruminants in the last decade, aiming to provide the new knowledge on DFM supplementation strategies for various ruminant production stages, and to identify what are the potential barriers and challenges for current ruminant industry to adopt the DFMs. Overall literature research indicates that DFMs have the potential to mitigate ruminal acidosis, improve immune response and gut health, increase productivity (growth and milk production), and reduce methane emissions or fecal shedding of pathogens. More research is needed to explore the mode of action of specific DFMs in the gut of ruminants, and the optimal supplementation strategies to promote the development and efficiency of DFM products for ruminants.

This experiment was conducted to investigate the effect of a spp.-based direct-fed microbial (DFM) on growth performance, apparent total tract digestibility (ATTD), blood profile, intestinal histomorphology, and fecal gas emission in piglets fed corn and soybean meal-based diets. The DFM product was based on 1 strain of and 2 strains of and formulated to supply 1.5 × 10 cfu/g of feed. A total of 128 piglets ([Yorkshire × Landrace] × Duroc; 6.8 ± 0.6 kg BW; weaning age: 24 d) were housed in groups (4 pigs/pen, 2 barrows and 2 gilts) and fed diets ( = 16) without or with DFM in a 2-phase feeding program: d 0 to 14 (phase I) and 15 to 42 (phase II). Feed intake and BW were measured weekly. At the end of each phase, samples for blood urea nitrogen (BUN), blood creatinine, ATTD, and fecal noxious gas emission were taken. At termination, 12 piglets per treatment were killed to access intestinal tissues for histomorphology. Overall, pigs fed DFM had a greater ( < 0.05) G:F than pigs fed the control diet. In phase I, pigs fed DFM showed a greater ( < 0.05) ADG and lower ( < 0.05) concentration of BUN and fecal ammonia emission than the control group. In phase II, a greater ( < 0.05) ATTD of nitrogen and longer ( < 0.05) duodenum and jejunum villi were observed in pigs fed the DFM diet compared with the control group. In conclusion, inclusion of DFM improved growth performance and villi length of the duodenum and jejunum in nursery pigs. Furthermore, DFM enhanced protein utilization as demonstrated by increased nitrogen digestibility, lower BUN, and lower fecal ammonia release.