O. Simon

Influence of xylanase-supplemented feed on the development of selected bacterial groups in the intestinal tract of broiler chicks

The colonization of Lactobacillus spp., enterobacteria and facultatively anaerobic gram-positive cocci was monitored in intestinal samples of growing broiler chicks from 24 h to 28 days of age. Rapid bacterial growth occurred within the first week, followed by stabilization and decline of colony forming units (CFU). Xylanase supplementation led to significantly lower CFU per gram of wet weight for total presumptive enterobacteria and total gram-positive cocci in luminal and tissue samples in the first 3 weeks. Lactobacillus spp. colony counts from tissue samples were higher for animals with the xylanase-supplemented diet, but luminal CFU were not. The composition of dominant Lactobacillus spp. strains was different in duodenal and jejunal tissues, but distribution of Lactobacillus spp. colony forms was unaffected by xylanase treatment. Mucosa-associated Enterococcus spp. displaced the dominant gram-positive cocci in the jejunal samples. d - and l -lactic acid and acetic acid concentrations were significantly higher in ileal samples from the control group on days 7 and 14, while butyric acid concentrations were higher in the xylanase-treated group. It is concluded that the less viscous intestinal environment caused by the xylanase slowed proliferation of gram-positive cocci and presumptive enterobacteria in enzyme-supplemented animals in the first 3 weeks of life.

Bacterial Responses to Different Dietary Cereal Types and Xylanase Supplementation in the Intestine of Broiler Chicken

Several studies were carried out to investigate the influence of dietary cereals differing in soluble non starch polysaccharides (NSP) content and a xylanase preparation on selected bacterial parameters in the small intestine of broiler chicken. Compared to a maize diet colony forming units (CFU) of mucosa associated bacteria were higher in a wheat/rye diet, most notably for enterobacteria and enterococci. Xylanase supplementation to the wheat/rye diet generally led to lower CFU, especially in the first week of life. However, xylanase supplementation also displayed higher in vitro growth potentials for enterobacteria and enterococci. Bacterial growth of luminal samples in minimal media supplemented with selected NSP showed that the wheat/rye diet enhanced bacterial capacities to utilize NSP only in ileal samples. The xylanase application generally shifted respective maximum growth to the proximal part of the small intestine. The presence of soluble NSP from wheat or rye in the diet per se did not enhance bacterial NSP hydrolyzing enzyme activities in the small intestine, but xylanase supplementation resulted in higher 1,3-1,4- g - glucanase activity. Compared to a maize diet the activity of bacterial bile salt hydrolases in samples of the small intestine was not increased due to inclusion of wheat/rye or triticale to the diet. However, xylanase supplementation led to a reduction with a corresponding increase of lipase activity. It was concluded that dietary cereals producing high intestinal viscosities lead to increased overall bacterial activity in the small intestine. The supplementation of a xylanase to cereal based diets producing high intestinal viscosity, changes composition and metabolic potential of bacterial populations and may specifically influence fat absorption in young animals.

Progression of mycotoxin and nutrient concentrations in wheat after inoculation with Fusarium culmorum.

The objective of this study was to follow the mycotoxin formation and changes in nutrient composition of wheat (cv. Ritmo) artificially inoculated with Fusarium culmorum. From anthesis until harvest, samples were taken once a week from the inoculated and control plots. The investigations were focused on monitoring the progression of the contamination of the wheat kernels with deoxynivalenol (DON) and zearalenone (ZON). Both the uncontaminated control kernels and the contaminated kernels were examined also for the presence of zearalenone-4-beta-D-glucopyranoside and several trichothecenes at harvest. Furthermore, the impact of the Fusarium inoculation on some nutrients as starch, crude protein, amino acid composition, crude ash, non starch polysaccharides (NSP) as well as viscosity and thousand seed weight (TSW) was examined. Also proteolytic and amylolytic activity as well as the NSP-degrading enzyme activities of inoculated and control samples were analysed at the time of harvest. DON was detected in higher concentrations and in earlier stages, while ZON was found later and in smaller amounts. On average 7.79 mg/kg DM of DON and 100 μg/kg DM of ZON were found in the inoculated kernels at the time of harvest. Neither in the contaminated nor in the control samples glucose conjugates of ZON (Zearalenone-4-beta-D-glucopyranoside) were detected. Moreover, the infection with Fusarium culmorum had pronounced effects on some quality parameters. The crude protein content of the inoculated kernels showed significantly higher values over the whole period compared to the control kernels. The protein content of the inoculated kernels amounted 13.9% DM at harvest, while only a concentration of 12.5% DM was detected in the control samples. Similarly, in almost all stages of development the crude ash content of inoculated samples was higher than in control samples. These distinct differences in kernel composition resulted possibly from the changes of the thousand seed weight. In the present work the grain harvested from the control plots showed a significantly higher TSW (24.2 g) as compared to their inoculated counterparts (15.5 g). Despite lower extract viscosity of inoculated samples at time of harvest, the content of soluble NSP of inoculated plots was higher than in control samples at the same time. Moreover, inoculation resulted in markedly increased activities of protease, amylase and several NSP-degrading enzyme activities. This would suggest that the cell wall penetrating properties of the fungus itself and/or that the fungus induced alterations of the metabolic activity of the embryo or other constituents of the wheat kernel could be responsible.

An investigation into the variability of extract viscosity of wheat-relationship with the content of non-starch-polysaccharide fractions and metabolisable energy for broiler chickens.

The in vitro extract-viscosity and the content of non-starch-polysaccharides were investigated in 34 defined wheat varieties grown at 5 locations each. Both, wheat genotype as well as growing location clearly influenced the viscosity of soluble extract from wheat. Furthermore, the content of non-starch-polysaccharides (soluble/total) and pentosans (soluble/total) were determined in 13 wheat varieties each grown at two locations. Soluble pentosan contents were highly positively correlated with extract viscosity of wheat at the locations Hayn (r = 0.86) and Biendorf (r = 0.90). The classical apparent metabolisable energy of 5 wheat samples having different extract viscosities was assessed. The AMEN values ranged from 14.0 to 14.6 MJ/kg DM and were significant negatively correlated to content of soluble arabinoxylans (r = 0.67) and to the extract viscosity (r = 0.83). Furthermore, the viscosity of jejunal (4.0 to 22.8 mPas) and ileal (13.1 to 78.0 mPas) digesta exhibited a clear relationship with soluble pentosan contents and extract viscosity. Under the conditions applied in this study the technique of extract viscosity measurement can predict the AME.

Comparative studies on the in vitro properties of phytases from various microbial origins.

The physical and chemical properties of six crude phytase preparations were compared. Four of these enzymes (Aspergillus A, Aspergillus R, Peniophora and Aspergillus T) were produced at commercial scale for the use as feed additives while the other two (E. coli and Bacillus) were produced at laboratory scale. The encoding genes of the enzymes were from different microbial origins (4 of fungal origin and 2 of bacterial origin, i.e., E. coli and Bacillus phytases). One of the fungal phytases (Aspergillus R) was expressed in transgenic rape. The enzymes were studied for their pH behaviour, temperature optimum and stability and resistance to protease inactivation. The phytases were found to exhibit different properties depending on source of the phytase gene and the production organism. The pH profiles of the enzymes showed that the fungal phytases had their pH optima ranging from 4.5 to 5.5. The bacterial E. coli phytase had also its pH optimum in the acidic range at pH 4.5 while the pH optimum for the Bacillus enzyme was identified at pH 7.0. Temperature optima were at 50 and 60°C for the fungal and bacterial phytases, respectively. The Bacillus phytase was more thermostable in aqueous solutions than all other enzymes. In pelleting experiments performed at 60, 70 and 80°C in the conditioner, Aspergillus A, Peniophora (measurement at pH 5.5) and E. coli phytases were more heat stable compared to other enzymes (Bacillus enzyme was not included). At a temperature of 70°C in the conditioner, these enzymes maintained a residual activity of approximately 70% after pelleting compared to approximately 30% determined for the other enzymes. Incubation of enzyme preparations with porcine proteases revealed that only E. coli phytase was insensitive against pepsin and pancreatin. Incubation of the enzymes in digesta supernatants from various segments of the digestive tract of hens revealed that digesta from stomach inactivated the enzymes most efficiently except E. coli phytase which had a residual activity of 93% after 60 min incubation at 40°C. It can be concluded that phytases of various microbial origins behave differently with respect to their in vitro properties which could be of importance for future developments of phytase preparations. Especially bacterial phytases contain properties like high temperature stability (Bacillus phytase) and high proteolytic stability (E. coli phytase) which make them favourable for future applications as feed additives.

Response of performance characteristics and fecal consistency to long-lasting dietary supplementation with the probiotic strain Bacillus cereus var. toyoi to sows and piglets

Abstract As part of an interdisciplinary research project, we studied the performance response of sows and their litters to the probiotic strain Bacillus cereus var. toyoi as well as feces consistency of piglets. Gestating sows (n = 26) were randomly allotted into two groups. The probiotic B. cereus var. toyoi was administered by dietary supplementation to one group of sows and their respective litters (probiotic group) whereas the second group (control group) received no probiotic supplementation. The duration of the application was nearly 17 weeks for sows (day 90 ante partum until day 28 post partum) and six weeks for piglets (day 15–56). Piglets were weaned after 28 days. Body weight and feed consumption were recorded weekly and fecal consistency of weaned piglets was studied daily. B. cereus var. toyoi was recovered from feces of sows and piglets as well as from digesta of piglets in the probiotic group, while being absent from all samples of control animals. In addition, the probiotic was detected in piglet feces and digesta before pre-starter feed was offered, indicating a second route of uptake besides diet. Sows of the probiotic group nursed numerically more piglets and supported a higher sum of total nursing days of all piglets within each litter than control sows (p = 0.04). In turn, body weight (BW) up to day 35 was greater for control piglets (p < 0.01), while average daily gain and gain to feed ratio (G:F) in weeks six and eight postweaning was higher in the probiotic group (p < 0.05). The overall G:F of the total postweaning period was 680 g/kg and 628 g/kg in the probiotic group and control group, respectively (p = 0.009). During the trial a high prevalence of liquid feces with its maximum in the second week after weaning was observed. Probiotic supplementation led to a reduction in the incidence of liquid feces and postweaning diarrhea by 38% and 59%, respectively (p < 0.001).

Growth behaviour of a spore forming probiotic strain in the gastrointestinal tract of broiler chicken and piglets

The growth behaviour of the probiotic strain Bacillus cereus var. toyoi in the gastrointestinal tract of broiler chicken and suckling piglets was evaluated. The strain germinated rapidly in intestinal samples from both animal species. Less than 10% of spores were recovered from the chicken crop and piglet stomach, respectively. Lumen samples and mucosal tissues from the hind gut of piglets displayed increasing colonization of the probiotic strain throughout the trial period. After oral administration of vegetative cells to broiler chicken and weaned piglets, sporulation was detected in all intestinal samples. The distribution of spore CFU indicated repeated germination and sporulation during the intestinal passage in piglets. B. cereus var. toyoi was not able to colonize the intestinal tract of both animal species. However, the probiotic strain was detected in suckling piglets before uptake of B. cereus var. toyoi supplemented feed. It is concluded that B. cereus var. toyoi germinates rapidly in broiler chicken and piglets, which is a necessary prerequisite for its possible probiotic effects. Germination and in vivo sporulation of vegetative cells indicated that the probiotic strain was metabolically active in the intestine of both animal species.

Interactions between dietary fat type and xylanase supplementation when rye‐based diets are fed to broiler chickens 2. Performance, nutrient digestibility and the fat‐soluble vitamin status of livers

1. The interactions between dietary fat type and xylanase supplementation of rye-based diets were investigated using a 2 x 2 factorial design in which a rye-based diet (610 g rye/kg) was combined with 100 g/kg of soya oil or beef tallow, with or without xylanase supplementation at 3000 IU/kg, and fed to 1-d-old male broilers for 35 d. Growth, nutrient digestibility and AMEN values were determined and the vitamin status of livers assessed at various ages. 2. There were significant interactions between crude fat and xylanase supplementation on the faecal digestibility of crude fat and crude protein. Fat digestibility was improved by xylanase in both fat type groups but to a greater extent for the tallow diets. However, the effects were found to be significant only for oleic, linoleic and linolenic acids. Similarly, protein digestibility and AMEN values were significantly improved by xylanase, but only for the tallow diet. 3. Ileal digestibility of nitrogen and amino acids also were affected by enzyme for both fat type diets, the effect generally being more pronounced for the tallow diet. 4. The deposition of the fat soluble vitamins A and E in livers was significantly increased by xylanase supplementation and was also better for soya oil than for tallow. 5. Xylanase supplementation increased the digestibility of insoluble pentosans whereas for the soluble pentosans the opposite effect was noted. 6. Without xylanase supplementation the tallow-based diet caused high mortality. Liveweight gain and the efficiency of food utilisation were greatest in the soya oil-based, xylanase-supplemented diet, followed by the unsupplemented soya oil-, supplemented tallow- and unsupplemented tallow-based diets.

In vitro and in vivo characteristics of bacterial phytases and their efficacy in broiler chickens

1. Three bacterial phytases derived from Bacillus, Escherichia coli or Klebsiella were compared with a phytase derived from Aspergillus niger in vitro and in vivo. 2. The in vitro results indicated that Aspergillus, E. coli and Klebsiella phytase displayed their activity optima in an acid pH range while Bacillus phytase did so in neutral pH. 3. The trials also revealed that only Bacillus phytase is more resistant to heat treatments, while E. coli and Klebsiella phytases are more stable against proteolytic inactivation. 4. In vivo phytases derived from Aspergillus, Bacillus, E. coli, Klebsiella or a combination of Bacillus and E. coli improved the utilisation of phosphorus (P balance) significantly to 0·54, 0·54, 0·55, 0·55 or 0·58, respectively, compared to 0·42 in the negative control. 5. The phytases used in this study seemed to be equally effective in improving P utilisation regardless of proposed intestinal site of activity. Combination of phytases acting in the gizzard with phytases acting in the intestine seems to be a promising way to further improving in vivo efficacy of phytases in poultry.

Effects of conjugated linoleic acids on protein to fat proportions, fatty acids, and plasma lipids in broilers

In a performance trial, broiler chickens received 29 g per kg feed of a preparation containing 70% linoleic acid (LA) in the control treatment and another preparation containing approximately the same amount of conjugated linoleic acids (CLA) in the experimental treatment. Diets of CLA treatment contained 18 g CLA per kg feed. The CLA preparation contained the isomers cis-9,trans-11 and trans-10,cis-12 at a proportion 1:1, other CLA isomers were quantitively negligible. Performance parameters (weight gain and feed conversion ratio over a 42 day period) were not significantly influenced by CLA intake. However, fat content of liver, breast, and leg muscles was reduced and protein contents in liver and leg muscles were elevated significantly. Fat to protein ratios in the main edible parts were shifted in favour of protein in CLA treated animals. In all analysed tissue lipids the content of saturated fatty acids was increased and that of monounsaturated fatty acids was decreased significantly. At the same time CLA was incorporated in tissue lipids effectively reaching more than 10 g per 100 g of total fatty acids. With regard to isomers the cis-9,trans-11 isomer was found in higher concentrations in tissue lipid fractions compared to the trans-10,cis-12 isomer. It was concluded that nutrient repartitioning due to CLA intake described for other species is also valid for broilers. Using appropriate feeding strategies it is possible to produce CLA enriched food from broilers.