Dietary Supplementation of a Commercial Prebiotic, Probiotic and Their Combination Affected Growth Performance and Transient Intestinal Microbiota of Red Drum (Sciaenops ocellatus L.).

In the present study, the potential synergism between beneficial lactic acid bacteria (Pediococcus acidilactici) contained in a probiotic and a mixture of fermentable complex carbohydrates and autolyzed brewer's yeast (or prebiotic) were explored in red drum. Four experimental diets were formulated from practical ingredients, and the basal diet was supplemented with either probiotic, prebiotic, or both supplements. Red drum juveniles (~5.5 g) were offered the four experimental diets for 56 days, and at the end of the feeding trial fish fed diets supplemented with probiotic had significantly better weight gain than those fed the non-supplemented diets, and higher protein content in their whole-body composition. Transient intestinal microbiome alpha and beta diversity were significantly affected by the dietary treatments. Interestingly, a higher relative abundance of the lactic acid genus Pediococcus was observed for fish fed diets supplemented with the prebiotic. A higher relative abundance was also observed for the predicted functions of the microbial metagenome, and many of these pathways involved the biosynthesis of essential amino acids, vitamins, and nucleotides. Even though no potential synergistic effect was observed, the individual inclusion of these prebiotic and probiotic supplements positively affected the intestinal health and growth performance of red drum, respectively.

Effect of moringa leaf powder and agave inulin on performance, intestinal morphology, and meat yield of broiler chickens.

The addition prebiotics in broiler diets can benefit digestion and nutrient abortion. The objective of the present study was to evaluate the effects of moringa leaf powder and agave inulin on growth performance, intestinal morphology, and slaughter traits of broiler chickens over 40 d of grow-out. A total of 280 broilers (Ross 308) aged 1 d were randomly allocated to 4 treatments, with 7 replicates each and 10 chicks per replicate: T1 = control diet, T2 = control diet with 15 g/kg of moringa leaf powder, T3 = control diet with 15 g/kg of agave inulin, and T4 = control diet with 15 g/kg of moringa leaf powder and 15 g/kg of agave inulin. The results showed that analysis of treatments at time were not different (P > 0.05) for broiler weights, feed and water intake, and weight gain. Treatment was significant (P < 0.05) for feed efficiency at 22 to 40 d; the T4 group presented higher (P < 0.05) values, and the T1 group presented lower (P < 0.05) values. However, the villus lengths of intestinal sections were different (P < 0.05) among treatments. In the duodenum, jejunum, and ileum, villus lengths were highest (P < 0.05) in the T2 group and lowest (P < 0.05) in the T3 and T4 groups. Villus widths in the duodenum and ileum were highest (P < 0.05) in the T2 group, but the T1 group showed highest (P < 0.05) values in the jejunum sections. The T3 and T4 groups showed lowest (P < 0.05) values in villus width in the duodenum, jejunum, and ileum. Thigh yield was highest (P < 0.05) in the T2 group and lowest (P < 0.05) in the T4 group. The T1 group exhibited the highest (P < 0.05) piece yields for leg, wing, and hip-back. The T4 group showed lowest (P < 0.05) leg and wing yields. Moringa leaf powder and agave inulin at a concentration of 15 g/kg in diets did not affect broiler performance, whereas moringa leaf powder improved intestinal morphology and thigh yield, and agave inulin improved leg yield. The results demonstrated benefits of these 2 feed additives to improve intestine health and meat yield in broilers over a 40-day grow-out.

Influence of sodium chlorate, ferulic acid, and essential oils on Escherichia coli and porcine fecal microbiota.

The influence of sodium chlorate (SC), ferulic acid (FA), and essential oils (EO) was examined on the survivability of two porcine diarrhetic enterotoxigenic Escherichia coli (ETEC) strains (F18 and K88) and populations of porcine fecal bacteria. Fecal bacterial populations were examined by denaturing gradient gel electrophoresis (DGGE) and identification by 16S gene sequencing. The treatments were control (no additives), 10 mM SC, 2.5 mg FA /mL, a 1.5% vol/vol solution of an EO mixture as well as mixtures of EO + SC, EO + FA, and FA + SC at each of the aforementioned concentrations. EO were a commercial blend of oregano oil and cinnamon oil with water and citric acid. Freshly collected porcine feces in half-strength Mueller Hinton broth was inoculated with E. coli F18 (Trial 1) or E. coli K88 (Trial 2). The fecal-E. coli suspensions were transferred to crimp top tubes preloaded with the treatment compounds. Quantitative enumeration was at 0, 6, and 24 h. All treatments reduced (P < 0.05) the counts of E. coli F18 at 6 and 24 h. With the exception of similarity coefficient (%SC), all the other treatments reduced (P < 0.05) the K88 counts at 24 h. The most effective treatments to reduce the F18 and K88 CFU numbers were those containing EO. Results of DGGE revealed that Dice percentage similarity coefficients (%SC) of bacterial profiles among treatment groups varied from 81.3% to 100%SC. The results of gene sequencing showed that, except for SC at 24 h, all the other treatments reduced the counts of the family Enterobacteriaceae, while Lactobacillaceae and Ruminococcaceae increased and Clostridiaceae decreased in all treatments. In conclusion, all treatments were effective in reducing the ETEC, but EO mixture was the most effective. The porcine microbial communities may be influenced by the studied treatments.

Nigella sativa L. as an alternative antibiotic feed supplement and effect on growth performance in weanling pigs.

BACKGROUND: Nigella sativa L. (NS) is a plant containing bioactive constituents such as thymoquinone. Extracts of NS improve performance and reduce enteropathogen colonization in poultry and small ruminants, but studies with swine are lacking. In two different studies oral administration of NS extracts at doses equivalent to 0, 1.5 and 4.5 g kg-1 diet was assessed on piglet performance and intestinal carriage of wildtype Escherichia coli and Campylobacter, and Salmonella Typhimurium. RESULTS: Wildtype E. coli populations in the jejunal and rectal content collected 9 days after treatment began were decreased (P ≤ 0.05). Populations recovered from pigs treated with extract at 1.5 and 4.5 g kg-1 diet were 0.72-1.31 log10 units lower than the controls (ranging from 6.05 to 6.61 log10 CFU g-1 ). Wildtype Campylobacter and Salmonella Typhimurium were unaffected by NS treatment. Feed efficiency over the 9 days improved linearly (P < 0.05) from 3.88 with 0 NS-treated pigs to 1.47 and 1.41 with pigs treated with NS at 1.5 and 4.5 g kg-1 diet, respectively, possibly due to high glutamine/glutamic acid content of the NS extract. CONCLUSION: NS supplementation of weanling pigs improved feed efficiency and helped control intestinal E. coli during this vulnerable production phase. © 2017 Society of Chemical Industry.

Molecular analysis of the caecal and tracheal microbiome of heat-stressed broilers supplemented with prebiotic and probiotic.

The gastrointestinal tract commensal microbiome is important for host nutrition, health and immunity. Little information is available regarding the role of these commensals at other mucosal surfaces in poultry. Tracheal mucosal surfaces offer sites for first-line health and immunity promotion in broilers, especially under stress-related conditions. The present study is aimed at elucidating the effects of feed supplementations with mannanoligosaccharides (MOS) prebiotic and a probiotic mixture (PM) on the caecal and tracheal microbiome of broilers kept under chronic heat stress (HS; 35 ± 2°C). Day-old chickens were randomly divided into five treatment groups: thermoneutral control (TN-CONT), HS-CONT, HS-MOS, HS-PM and HS synbiotic (fed MOS and PM). Caecal digesta and tracheal swabs were collected at day 42 and subjected to DNA extraction, followed by polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) and pyrosequencing. The PCR-DGGE dendrograms revealed significant (49.5% similarity coefficients) differences between caecal and tracheal microbiome. Tracheal microbiome pyrosequencing revealed 9 phyla, 17 classes, 34 orders, 68 families and 125 genera, while 11 phyla, 19 classes, 34 orders, 85 families and 165 genera were identified in caeca. An unweighted UniFrac distance metric revealed a distinct clustering pattern (analysis of similarities, P = 0.007) between caecal and tracheal microbiome. Lactobacillus was the most abundant genus in trachea and caeca and was more abundant in caeca and trachea of HS groups compared with the TN-CONT group. Distinct bacterial clades occupied the caecal and tracheal microbiomes, although some bacterial groups overlapped, demonstrating a core microbiome dominated by Lactobacillus. No positive effects of supplementations were observed on abundance of probiotic bacteria.

Evaluation of linalool, a natural antimicrobial and insecticidal essential oil from basil: effects on poultry.

Linalool is a natural plant-product used in perfumes, cosmetics, and flavoring agents. Linalool has proven antimicrobial and insect-repellent properties, which indicate it might be useful for control of enteropathogens or insect pests in poultry production. However, there are no published reports that linalool may be safely administered to or tolerated by chickens. Linalool was added to the diets of day-of-hatch chicks, and they were fed linalool-supplemented diets for 3 wk. We studied the effects of linalool on serum chemistry, gross pathology, feed conversion, and relative liver weights. Linalool had a dramatic negative dose-dependent effect on feed conversion at concentrations in the feed exceeding 2% linalool, but not on gross pathology. Liver weights were significantly increased in the 5% linalool-treated birds. There was a statistical effect on blood glucose, but this parameter remained below the cut-offs for elevated serum glucose, and the result is likely of no biological significance. Linalool caused serum aspartate aminotransferase (AST) levels to increase, but it did not increase serum gamma-glutamyl transferase levels. The linalool effect on AST was dose-dependent, but in linalool doses between 0.1 and 2% of the feed, AST was not elevated beyond normal parameters. Linalool at 2% or less may be safely added to chicken feed. We suggest future studies to evaluate the addition of linalool to the litter, where it may be used as an antimicrobial or an insect repellent or to produce a calming effect.

Molecular identification and characterization of ileal and cecal fungus communities in broilers given probiotics, specific essential oil blends, and under mixed Eimeria infection.

Broiler digestive tract fungal communities have gained far less scrutiny than that given corresponding bacterial communities. Attention given poultry-associated fungi have focused primarily on feed-associated toxin-producers, yeast, and yeast products. The current project focused on the use of pyrosequencing and denaturing gradient gel electrophoresis (DGGE) to identify and monitor broiler digestive fungal communities. Eight different treatments were included. Four controls were an Uninfected-Unmedicated Control, an Unmedicated-Infected Control, the antibiotic bacitracin methylene disalicylate plus the ionophore monensin as Positive Control, and the ionophore monensin alone as a Negative Control. Four treatments were two probiotics (BC-30 and Calsporin) and two specific essential oil blends (Crina Poultry Plus and Crina Poultry AF). All chickens except the Unmedicated-Uninfected Control were given, at 15 days of age, a standard oral Eimeria inoculum of sporulated oocysts. Ileal and cecal digesta were collected at pre-Eimeria infection at 14 days of age and at 7 days post-Eimeria infection at 22 days of age. Extracted cecal DNA was analyzed by pyrosequencing to examine the impact of diet supplements and Eimeria infection on individual constituents in the fungal community, while DGGE was used to compare more qualitative changes in ileal and cecal communities. Pyrosequencing identified three phyla, seven classes, eight orders, 13 families, 17 genera, and 23 fungal species. Ileal and cecal DGGE patterns showed fungal communities were clustered mainly into pre- and post-infection patterns. Post-infection Unmedicated-Uninfected patterns were clustered with pre-infection groups demonstrating a strong effect of Eimeria infection on digestive fungal populations. These combined techniques offered added versatility towards unraveling the effects of enteropathogen infection and performance enhancing feed additives on broiler digestive microflora.

Effect of chlorate, molybdate, and shikimic acid on Salmonella enterica serovar Typhimurium in aerobic and anaerobic cultures.

Experiments were conducted to determine factors that affect sensitivity of Salmonella enterica serovar Typhimurium to sodium chlorate (5mM). In our first experiment, cultures grown without chlorate grew more rapidly than those with chlorate. An extended lag before logarithmic growth was observed in anaerobic but not aerobic cultures containing chlorate. Chlorate inhibition of growth during aerobic culture began later than that observed in anaerobic cultures but persisted once inhibition was apparent. Conversely, anaerobic cultures appeared to adapt to chlorate after approximately 10h of incubation, exhibiting rapid compensatory growth. In anaerobic chlorate-containing cultures, 20% of total viable counts were resistant to chlorate by 6h and had propagated to 100% resistance (>10(9)CFU mL(-1)) by 24h. In the aerobic chlorate-containing cultures, 12.9% of colonies had detectable resistance to chlorate by 6h, but only 1% retained detectable resistance at 24h, likely because these cultures had opportunity to respire on oxygen and were thus not enriched via the selective pressure of chlorate. In another study, treatment with shikimic acid (0.34 mM), molybdate (1mM) or their combination had little effect on aerobic or anaerobic growth of Salmonella in the absence of added chlorate. As observed in our earlier study, chlorate resistance was not detected in any cultures without added chlorate. Chlorate resistant Salmonella were recovered at equivalent numbers regardless of treatment after 8h of aerobic or anaerobic culture with added chlorate; however, by 24h incubation chlorate sensitivity was completely restored to aerobic but not anaerobic cultures treated with shikimic acid or molybdate but not their combination. Results indicate that anaerobic adaptation of S. Typhimurium to sodium chlorate during pure culture is likely due to the selective propagation of low numbers of cells exhibiting spontaneous resistance to chlorate and this resistance is not reversible by molybdenum supplementation.

Dietary supplementation of short-chain fructooligosaccharides influences gastrointestinal microbiota composition and immunity characteristics of Pacific white shrimp, Litopenaeus vannamei, cultured in a recirculating system.

Supplementation of prebiotic compounds, including short-chain fructooligosaccharides (scFOS) has been shown to confer benefits on nutrient utilization, growth, and disease resistance of various animal species through improved gastrointestinal (GI) microbiota. However, potential uses of prebiotics for shrimp have not been defined. A 6-wk feeding trial was conducted in a recirculating system to determine the effects of scFOS supplementation on growth performance, immune functions, and GI microbiota composition of Pacific white shrimp (Litopenaeus vannamei). scFOS was supplemented in a nutritionally complete diet (35% crude protein) at 0.025, 0.0500, 0.075, 0.100, 0.200, 0.400, and 0.800% by weight. After 6 wk of feeding, shrimp fed 0, 0.1, and 0.8% scFOS were sampled for assays of immune function and GI microbiota. Dietary supplementation of scFOS did not improve weight gain, feed conversion ratio, or survival of shrimp. Denaturing gradient gel electrophoresis analysis suggested the intestinal tract microbial community from shrimp fed the basal diet was different from that of shrimp fed the scFOS diets [similarity coefficient (SC) = 74.9%)], although the intestinal tract microbial community from shrimp fed the scFOS-supplemented diets was very similar (SC = 92.3%). All the bacterial species contributing to the GI microbial differences were identified, although most of them are uncultured species. Both total hemocyte count and hemocyte respiratory burst increased (P < 0.05) by incremental dietary supplementation of scFOS (0-0.8%). This study is the first to our knowledge to show that dietary scFOS can selectively support growth of certain bacterial species in the GI tract of shrimp and enhance immunity, which may facilitate development of alternative strategies, including novel probiotics and synbiotics, for shrimp growth and health management.