Short-term exposure to fumonisins and deoxynivalenol, on broiler growth performance and cecal Salmonella load during experimental Salmonella Enteritidis infection.

Fumonisins (FUM) and deoxynivalenol (DON) are two common mycotoxins in poultry feed. Salmonella enterica ser. Enteritidis (S. Enteritidis) is a primary foodborne bacterium in broilers. This trial was conducted to evaluate the effects of naturally occurring FUM and DON and their combination at subclinical doses on broiler performance during a S. Enteritidis challenge. The experiment consisted of five treatments: NCC, no-challenge no-mycotoxin treatment; CC, Salmonella challenge + no-mycotoxin treatment; DON, DON 0.6 mg/kg + Salmonella challenge; FUM, FUM 14 mg/kg + Salmonella challenge; DON + FUM + T-2 + neosolaniol, DON 0.6 mg/kg + FUM 14 mg/kg + T-2 toxin 0.6 mg/kg + 0.8 mg/kg neosolaniol + Salmonella challenge. On d 4, birds were challenged with either 0 or 1 × 109 CFU/mL S. Enteritidis orally. There were no significant effects on growth performance among treatments at 0, 3, 7, and 14 d of post-inoculation (dpi). On 14 dpi, the combined DON + FUM + T-2 + neosolaniol significantly increased the Salmonella load by 1.5 logs compared to the control groups (P < 0.05). FUM significantly increased the cecal tonsil IL-10 gene expression by 1.2-fold at 7 dpi (P < 0.05) and downregulated TNF-α by 1.8-fold on 14 dpi compared to the control, nonchallenge groups (P < 0.05). On 7 dpi, the combined DON + FUM + T-2 + neosolaniol reduced occludin by 4.4-fold (P < 0.05) when compared to the control groups. Similarly, combined DON + FUM+ T-2 + neosolaniol decreased zona-occluden transcription by 2.3 and 7.6-fold on 3 and 14 dpi, respectively (P < 0.05). Furthermore, combined DON + FUM + T-2 + neosolaniol decreased Claudin-1 by 2.2-fold and Claudin-4 by 5.1-fold on 14 dpi when compared to the control groups (P < 0.05). In conclusion, short-term exposure to a subclinical dose of combined DON + FUM + T-2 + neosolaniol had an impact on broiler intestinal tight junction proteins and cecal Salmonella abundance under experimental Salmonella challenge.

Subclinical Doses of Combined Fumonisins and Deoxynivalenol Predispose Clostridium perfringens-Inoculated Broilers to Necrotic Enteritis.

Fumonisins (FB) and deoxynivalenol (DON) are mycotoxins which may predispose broiler chickens to necrotic enteritis (NE). The objective of this study was to identify the effects of subclinical doses of combined FB and DON on NE. A total of 480 day-old male broiler chicks were divided into four treatment groups; 1) control group (basal diet + Clostridium perfringens); 2) necrotic enteritis group (basal diet + Eimeria maxima + C. perfringens); 3) FB + DON group (basal diet + 3 mg/kg FB + 4 mg/kg DON + C. perfringens); and 4) FB + DON + NE group (basal diet + 3 mg/kg FB + 4 mg/kg DON + E. maxima + C. perfringens). Birds in NE and FB + DON + NE groups received 2.5 × 103 E. maxima on day 14. All birds were inoculated with C. perfringens on days 19, 20, and 21. On day 35, birds in the NE, FB + DON, and FB + DON + NE groups had 242, 84, and 339 g lower BWG and a 19-, 2-, and 22-point increase in FCR respectively, than in the control group. Subclinical doses of FB + DON increased (p < 0.05) the NE lesion scores compared to the control group on day 21. On day 21, birds in the NE, FB + DON, and FB + DON + NE groups had increased (p < 0.05) serum FITC-D, lower (p < 0.05) jejunal tight junction protein mRNA, and increased (p < 0.05) cecal tonsil IL-1 mRNA compared to control group. On day 21, birds in the NE group had decreased (p < 0.05) villi height to crypt depth ratio compared to the control group and the presence of FB + DON in NE-induced birds further decreased the villi height to crypt depth ratio. Birds in the NE, FB + DON, and FB + DON + NE groups had increased (p < 0.05) C. perfringens, lower (p < 0.05) Lactobacillus loads in the cecal content, and a lower (p < 0.05) CD8+: CD4+ cell ratio in the cecal tonsils compared to the control group. It can be concluded that subclinical doses of combined FB and DON predispose C. perfringens-inoculated birds to NE, and the presence of FB + DON in NE-induced birds exacerbated the severity of NE.

Effect of a synbiotic supplement as an antibiotic alternative on broiler skeletal, physiological, and oxidative parameters under heat stress.

The aim of this study was to examine if synbiotics can function as alternatives to antibiotics in broiler production under heat stress (HS). Day-old broiler chicks (528 birds) were randomly placed in floor pens within 2 identical temperature-controlled rooms (11 birds/pen and 24 pens/room). The pens of each room were evenly divided among 3 treatments (n = 8): basal diet (CON), the basal diet mixed with 50 ppm of bacitracin methylene disalicylate (BMD) or a synbiotic (50 ppm of PoultryStar meUS, SYN). From d 15, room 2 was under thermoneutral (TN) conditions (TN-CON, TN-BMD, and TN-SYN), while HS was applied to room 1 at 32oC for 9 hrs/d (0800 to 1700) (HS-CON, HS-BMD, and HS-SYN). Treatment effects on footpad dermatitis and gait score were measured on 5 birds/pen, and latency to lie (LTL) test was measured on 2 birds/pen at d 27 and d 41; and 1 broiler/pen was sampled on d 28 and d 42, respectively. Body, liver, and spleen weight were determined. Plasma levels of interleukins (IL), heat shock protein 70, immunoglobulin (Ig)Y, liver superoxide dismutase (SOD) and glutathione peroxidase (GPx) enzyme activities were examined. Heat stress suppressed BW and IgY concentrations on both d 28 and d 42, while suppressed plasma IL-6 concentrations, SOD activities, and LTL duration on d 28 only (P < 0.05). Among all treatments, SYN birds had the best foot and skeletal health scores on both d 27 and d 41 (P < 0.05). On d 42, SYN increased BW, and TN-SYN birds had higher relative spleen weight than both TN-BMD and TN-CON birds (P < 0.05). Antibiotic BMD increased BW (P < 0.05) but decreased SOD activities (P < 0.05) on d 42. These results indicate that the SYN supplementation decreases HS negative effect on broilers by improving BW, foot, and skeletal health, while BMD improves BW but also increases oxidative stress in broilers. The data suggest that synbiotic supplement may function as an alternative to antibiotics in broiler production during summer seasons, especially in the tropical and subtropical regions.

The impact of deoxynivalenol, fumonisins, and their combination on performance, nutrient, and energy digestibility in broiler chickens.

This study evaluated the effects of the mycotoxins deoxynivalenol (DON), fumonisins (FUM), and their combination on growth performance, nutrient, and energy digestibility in broilers. A total of 960 Cobb-Cobb male broilers were obtained on the day of hatch and placed 10 birds per cage with 8 cages per treatment. The experiment consisted of 12 treatments: control; DON 1.5 mg/kg; DON 5.0 mg/kg; FUM 20.0 mg/kg; DON 1.5 mg/kg + FUM 20.0 mg/kg; and DON 5.0 mg/kg + FUM 20 mg/kg. The remaining dietary treatments were the correlative nitrogen-free diets (NFD) for determining the endogenous nutrients loss. All birds were fed with a corn-soybean meal diet from days 1 to 15, until birds from latter 6 treatments were switched to their correlative NFD diet from days 15 to 21. Feed and BW were weighed by cage on days 8, 15, and 21. On day 21, ileal digesta was collected for digestibility determination. Both DON 1.5 mg/kg + FUM 20 mg/kg and DON 5.0 mg/kg + FUM 20 mg/kg treatments showed reduced feed intake (P ≤ 0.05) from days 8 to 15 and days 15 to 21. However, no significant effects were noted for BW gain or mortality-adjusted feed conversion ratio after adding single or combined mycotoxin on days 8 and 15. At day 21, cumulative BW gain was less (P ≤ 0.05) in birds fed with the mycotoxin combination diets than the control. No significant changes were shown for ileal endogenous amino acids losses. Control treatment had significantly higher (P ≤ 0.05) apparent ileal energy digestibility than the DON 5.0 mg/kg + FUM 20.0 mg/kg treatment (3,126 vs. 2,895 kcal/kg), representing a 5%-unit loss in apparent DM digestibility. No significant difference was found for standardized crude protein and amino acid digestibility. In conclusion, the combination of DON and FUM (DON 1.5 mg/kg + FUM 20 mg/kg or DON 5.0 mg/kg + FUM 20 mg/kg) reduced DM and ileal energy digestibility, which negatively affected BW gain in broilers.

Research Note: Effect of synbiotic supplementation on caecal Clostridium perfringens load in broiler chickens with different necrotic enteritis challenge models.

Studies were conducted to determine the efficacy of synbiotic applications to combat the negative effects of necrotic enteritis (NE). An in vitro study was conducted to test the effect of probiotics species supernatants to decrease Clostridium perfringens (CP) proliferation. Lactobacillus reuteri, Enterococcus faecium, Bifidobacterium animalis, and Pediococcus acidilactici culture supernatants decreased the proliferation of CP at 1:1 supernatant-to-pathogen dilution in vitro. Two in vivo studies were conducted to determine the in vivo response of synbiotic supplementation containing the aforementioned probiotic strains on broiler production performance and caecal CP load in broilers induced with NE infection. In experiment 1, 75 broiler chicks were randomly allotted to 3 treatment groups, control (basal diet), ionophore (Salinomycin), and synbiotic (PoultryStar me), from day of hatch, and NE was induced in all birds. There were no significant treatment effects on BW, feed consumption, and feed gain ratio. However, at 35 D, ionophore or synbiotic supplementation increased (P < 0.05) villi height and decreased interleukin (IL)-1 mRNA abundance, while synbiotic supplementation increased (P < 0.05) IL-10 mRNA abundance compared with the control group, respectively. In experiment 2, 360 broiler chicks were randomly allotted to 3 treatments, an unchallenged negative control (control; basal diet), challenged positive control (NE; basal diet), or NE + synbiotic group (synbiotic). At both 21 and 42 D of age, NE birds had decreased (P < 0.05) BW, feed conversion, and jejunal villi height compared with control, while NE + synbiotic birds were not different from control groups. At 42 D of age, NE birds had 2.2 log/g increased CP in the ceca contents compared with control, while synbiotic birds had CP load that was not different than that of the control group. NE + synbiotic birds had significantly greater amounts of bile anti-CP IgA than the control and NE groups. It can be concluded that synbiotic supplementation decreased CP proliferation in vitro and caecal CP load in vivo while improving production parameters during an NE infection in broilers.

Synbiotic supplementation to decrease Salmonella colonization in the intestine and carcass contamination in broiler birds.

In vitro and in vivo experiments were conducted to study the effects of synbiotic supplementation on Salmonella enterica ser. Enteritidis (SE) proliferation, cecal content load, and broiler carcass contamination. Lactobacillus reuteri, Enterococcus faecium, Bifidobacterium animalis, and Pediococcus acidilactici culture supernatants decreased (P < 0.05) the in vitro proliferation of SE at 1:1 supernatant: pathogen dilution. A total of 240 Cobb-500 broiler chicks were randomly allotted to three treatment groups (8 replicates/group with 10 birds/replicate): control (basal diet), antibiotic (Virginiamycin at 20 mg/kg feed), synbiotic (PoultryStar® ME at 0.5 g/kg feed containing L. reuteri, E. faecium, B. animalis, P. acidilactici and a Fructooligosaccharide) from day of hatch. At 21 d of age, all birds in experimental groups were orally inoculated with 250 µl of 1 X 109 CFU SE. Antibiotic supplementation increased (P < 0.05) body weight and feed consumption, compared to the control group. Birds in the synbiotic supplementation had intermediate body weight and feed consumption that were not significantly different from both the control and antibiotic group at 42 d of age in SE infected birds. No significant effects were observed in feed efficiency at 42 d of age among the groups. Antibiotic and synbiotic supplementation decreased (P < 0.05) SE load in cecal contents by 0.90 and 0.85 log units/ g and carcass SE load by 1.4 and 1.5 log units/mL of rinsate compared to the control group at 42 d of age (21 dpi). The relative abundance of IL-10, IL-1, TLR-4, and IFNγ mRNA was decreased (P < 0.05) in the antibiotic and synbiotic supplementation groups compared to the control birds at 42 d of age (21 dpi). It can be concluded that synbiotic supplementation decreased SE proliferation in vitro and decreased SE load in the cecal contents and broiler carcass.

Effects of probiotic supplementation on performance traits, bone mineralization, cecal microbial composition, cytokines and corticosterone in laying hens.

Recent researches have showed that probiotics promote bone health in humans and rodents. The objective of this study was to determine if probiotics have the similar effects in laying hens. Ninety-six 60-week-old White Leghorn hens were assigned to four-hen cages based on their BW. The cages were randomly assigned to 1 of 4 treatments: a layer diet mixed with a commercial probiotic product (containing Enterococcus faecium, Pediococcus acidilactici, Bifidobacterium animalis and Lactobacillus reuteri) at 0, 0.5, 1.0 or 2.0 g/kg feed (Control, 0.5×, 1.0× and 2.0×) for 7 weeks. Cecal Bifidobacterium spp. counts were higher in all probiotic groups (P0.05). In addition, the plasma concentrations of cytokines (interleukin-1ß, interleukin-6, interleukin-10, interferon-γ and tumor necrosis factor-α) and corticosterone as well as the levels of heterophil to lymphocyte ratio were similar between the 2.0× group and the control group (P>0.05). In line with these findings, no differences of cecal tonsil mRNA expressions of interleukin-1ß, interleukin-6 and lipopolysaccharide-induced tumor necrosis factor-α factor were detected between these two groups (P>0.05). These results suggest that immune cytokines and corticosterone may not involve in the probiotic-induced improvement of eggshell quality and bone mineralization in laying hens. In conclusion, the dietary probiotic supplementation altered cecal microbiota composition, resulting in reduced shell-less egg production and improved bone mineralization in laying hens; and the dietary dose of the probiotic up to 2.0× did not cause negative stress reactions in laying hens.

Effects of a dietary synbiotic inclusion on bone health in broilers subjected to cyclic heat stress episodes.

The objective of this study was to determine the effects of a dietary synbiotic inclusion on broiler bone health under daily cyclic heat stress. A total of 360 Ross 708 broilers were randomly assigned to 1 of 3 dietary treatments (8 replicates per treatment): a regular diet (control) and the regular diet mixed with a commercial synbiotic product at 0.5 (0.5X) or 1.0 (1.0X) g/kg. The synbiotic contains a prebiotic (fructooligosaccharides) and a probiotic mixture of 4 microbial strains (Enterococcus faecium, Pediococcus acidilactici, Bifidobacterium animalis, and Lactobacillus reuteri). Room temperature was gradually decreased from 34°C on d 1 by 0.5°C/d for the first 14 d; then a cyclic heat stress episode (32°C/9 h/d) was applied from d 15 to 42. Gait score assessment and the latency-to-lie test were conducted when broilers were 40 and 41 d of age, respectively. The tibia, femur, and humerus were collected for measuring bone parameters at 42 d of age. The data indicated that bone mineral density, bone mineral content, and bone area were higher and the level of gait score was lower in the 1.0X group (P = 0.05) but not in the 0.5X group (P > 0.05) compared to controls. The proportions of broilers showing signs of lameness were ranked 1.0X group (25%) < 0.5X group (45%) < control (54%). Compared to controls, broilers of 0.5X group stood longer (P = 0.03) during the latency-to-lie test. In conclusion, under the present conditions the synbiotic profoundly improves multiple indices of leg health of broilers subjected to the cyclic heat episodes, resulting in an improvement in walking ability.

Effect of dietary synbiotic supplement on behavioral patterns and growth performance of broiler chickens reared under heat stress.

This study examined the effects of a dietary synbiotic supplement on the behavioral patterns and growth performance of broiler chickens exposed to heat stress (HS). Three hundred sixty 1-day-old male Ross 708 broiler chicks were distributed among 24 floor pens (15 chicks per pen); each pen was randomly assigned to one of 3 dietary treatments containing a synbiotic at 0 (control), 0.5 (0.5X) and 1.0 (1.0X) g/kg. From d 15 to 42, birds were exposed to HS at 32°C daily from 08:00 to 17:00. Five broiler chickens were randomly marked in each pen for behavioral observation. Instantaneous scan sampling was used to record the birds' behavioral patterns. Performance parameters were measured on d 7, 14, 28 and 42. The synbiotic fed birds exhibited more standing, sitting, walking, feeding, preening and less wing spreading and panting behaviors (P < 0.05) compared to birds fed the control diet. The synbiotic group also had higher BW, BW gain and feed intake on d 7, 14 and 42 (P < 0.05), and higher BW, feed intake and feed conversion ratio at d 28 (P < 0.01). There were no treatment effects on drinking behavior, BW gain on d 28 and feed conversion ratio on d 42 (P > 0.05). There were few dose-related differences of the synbiotic on production performance; namely, the 1.0X concentration resulted in the highest BW and feed intake on d 14 and 42 (P < 0.05), while BW gain was higher compared to the control group only on d 42 (P < 0.05). The results suggest that the synbiotic supplement may prove to be an important management tool for the broiler industry to diminish the negative effects of HS, potentially safeguarding the welfare and production of broiler chickens, particularly in areas that experience hot climates.

Effect of synbiotic supplementation on layer production and cecal Salmonella load during a Salmonella challenge.

This study analyzed the inhibitory effects of a synbiotic product (PoultryStar® me) on production parameters, intestinal microflora profile, and immune parameters in laying hens with and without a Salmonella challenge. The synbiotic product contained 4 probiotic bacterial strains (Lactobacillus reuteri, Enterococcus faecium, Bifidobacterium animalis, and Pediococcus acidilactici) and a prebiotic fructooligosaccharide. Layers were supplemented with the synbiotic from d of hatch to 28 wk of age. At 16 wk of age, birds were either vaccinated with Salmonella enterica Enteritidis (SE) vaccine or left unvaccinated. At 24 wk of age, a portion of the birds was challenged with 1 × 109 CFU of SE or left unchallenged, resulting in a 3 (vaccinated, challenged, or both vaccinated and challenged) X 2 (control and synbiotics) factorial arrangement of treatments. At 18 and 20 wk of age, birds fed synbiotics in both vaccinated and unvaccinated groups had increased (P < 0.05) BW more than those in the un-supplemented groups. Birds fed synbiotics had 0.7, 17.8, 21.7, 3, and 4.2% higher (P < 0.05) hen d egg production (HDEP) at 19, 20, 21, 22, and 23 wk of age, compared to the birds without supplementation, respectively. After administering the SE challenge, supplemented birds had 3, 6.7, 4.3, 12.5, and 14.4% higher (P < 0.05) HDEP at 24, 25, 26, 27, and 28 wk of age, compared to the birds not supplemented, respectively. Irrespective of the vaccination status, birds fed synbiotics and challenged with SE had a lower (P < 0.05) SE cecal load compared to the un-supplemented groups. At 22 d post Salmonella challenge, birds supplemented, vaccinated, and challenged had the highest bile IgA content. It can be concluded that supplementation of the synbiotic product could be beneficial to layer diets as a growth promoter, performance enhancer, and for protection against SE infection.

Prevalence and effects of mycotoxins on poultry health and performance, and recent development in mycotoxin counteracting strategies.

Extensive research over the last couple of decades has made it obvious that mycotoxins are commonly prevalent in majority of feed ingredients. A worldwide mycotoxin survey in 2013 revealed 81% of around 3,000 grain and feed samples analyzed had at least 1 mycotoxin, which was higher than the 10-year average (from 2004 to 2013) of 76% in a total of 25,944 samples. The considerable increase in the number of positive samples in 2013 may be due to the improvements in detection methods and their sensitivity. The recently developed liquid chromatography coupled to (tandem) mass spectrometry allows the inclusion of a high number of analytes and is the most selective, sensitive, and accurate of all the mycotoxin analytical methods. Mycotoxins can affect the animals either individually or additively in the presence of more than 1 mycotoxin, and may affect various organs such as gastrointestinal tract, liver, and immune system, essentially resulting in reduced productivity of the birds and mortality in extreme cases. While the use of mycotoxin binding agents has been a commonly used counteracting strategy, considering the great diversity in the chemical structures of mycotoxins, it is very obvious that there is no single method that can be used to deactivate mycotoxins in feed. Therefore, different strategies have to be combined in order to specifically target individual mycotoxins without impacting the quality of feed. Enzymatic or microbial detoxification, referred to as "biotransformation" or "biodetoxification," utilizes microorganisms or purified enzymes thereof to catabolize the entire mycotoxin or transform or cleave it to less or non-toxic compounds. However, the awareness on the prevalence of mycotoxins, available modern techniques to analyze them, the effects of mycotoxicoses, and the recent developments in the ways to safely eliminate the mycotoxins from the feed are very minimal among the producers. This symposium review paper comprehensively discusses the above mentioned aspects.

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.

Validation of the effects of small differences in dietary metabolizable energy and feed restriction in first-cycle laying hens.

An experiment was conducted to evaluate energy utilization of laying hens fed diets containing 2 ME concentrations, using response criteria including performance, BW, abdominal fat pad, and energy digestibility. The experiment was a 2 × 2 factorial with 2 feeding regimens (ad libitum and restriction fed), and 2 dietary ME levels [2,880 kcal/kg of ME (CON); and 2,790 kcal/kg of ME (LME)]. A total of 60 Hy-Line W36 first-cycle laying hens were fed experimental diets, resulting in 15 individually caged hens for each of the 4 treatments. Hens in the restriction-fed group were fed 90 g of feed per day. The CON diet was formulated to meet or exceed the NRC (1994) recommendations with 2,880 kcal/kg, whereas the LME diet was similar with the exception of a 90 kcal/kg reduction in ME. Hens were fed experimental diets for 12 wk from hen 28 to 39 wk of age. Hen day egg production, weekly feed intake, and every 2 wk, egg weights and egg mass were recorded, whereas hen BW was measured every 4 wk. Excreta samples were collected over the last 5 d of experiment to determine AMEn. Abdominal fat pads were measured individually for all hens at the end of experiment. There were no interactions between feeding regimens and dietary ME levels throughout the experiment. Feed restriction resulted in reductions (P ≤ 0.01) in hen day egg production, BW, and abdominal fat pad, indicating reduced nutrient availability to partition toward production, maintenance, and storage functions. The reduction in energy intake between CON and LME fed birds (90 kcal/kg) did not change the energy partitioned toward production or maintenance, but reduced (P = 0.03) the energy stored (reduced fat pad) of LME-fed hens. These results suggest that energy is used following the pattern of production and maintenance before storage requirements and that fat pad (energy storage) may be the most sensitive indicator of dietary energy status over short-term in Hy-Line W36 laying hens.