Ricarda M. Engberg

Effects of Dietary Fat Source and Subtherapeutic Levels of Antibiotic on the Bacterial Community in the Ileum of Broiler Chickens at Various Ages

ABSTRACT The effect of dietary fat source (soy oil or a mixture of lard and tallow) and dietary supplementation with antibiotics (a combination of avilamycin at 10 mg kg of feed−1 and salinomycin at 40 mg kg of feed−1) on the bacterial community in the ileum of broiler chickens at different ages (7, 14, 21, and 35 days) was studied using PCR with denaturing gradient gel electrophoresis (DGGE) analysis and bacteriological culture. The bacterial origin of fragments in DGGE profiles was identified by sequencing. Bacterial enumeration results, together with PCR-DGGE profiles, showed that the composition of the microflora was age dependent and influenced by dietary fat source and antibiotic supplementation. An increased incidence of streptococci, enterobacteria, and Clostridium perfringens with age of the chickens was demonstrated. Lactobacilli and C. perfringens were the bacterial groups most strongly affected by the dietary treatments. Moreover, different strains (clonal variants of the alpha-toxin gene) of C. perfringens type A were detected in response to age, dietary fat source, and dietary supplementation with antibiotics.

The influence of grinding and pelleting of feed on the microbial composition and activity in the digestive tract of broiler chickens

1. The influence of feed grinding (coarsely or finely ground feed) and feed form (mash or pellets) on the intestinal environment was investigated in a growth experiment with broiler chickens taking the intestinal microflora, intestinal viscosity, and the activities of pancreatic digestive enzymes into consideration. 2. As compared to mash the feeding of pellets was associated with a significantly higher body weight due to increased feed intake and improved feed utilisation. 3. Pellet-fed birds had significantly decreased gizzard weights, a higher gizzard pH and a lower intestinal pH than mash-fed birds. 4. Pellet-fed birds had significantly lower relative pancreas weights and lower activities of pancreatic digestive enzymes (amylase, lipase, chymotrypsin), which indicates the existence of a feedback mechanism, which may have been triggered by the intestinal concentration of enzymatically hydrolysed products or of the respective digestive enzymes. 5. Pellet-fed birds had larger numbers of coliform bacteria and enterococci in the ileum and a reduced number of Clostridium perfringens and lactobacilli in the distal end of the digestive tract (caeca and rectum). Microbial fermentation in terms of volatile fatty acid (VFA) concentration was found to be significantly higher in the caeca of pellet-fed birds than in mash-fed birds.

Methylotrophic methanogenic Thermoplasmata implicated in reduced methane emissions from bovine rumen

Rumen methanogens are major sources of anthropogenic methane emissions, and these archaea are targets in strategies aimed at reducing methane emissions. Here we show that the poorly characterised Thermoplasmata archaea in bovine rumen are methylotrophic methanogens and that they are reduced upon dietary supplementation with rapeseed oil in lactating cows. In a metatranscriptomic survey, Thermoplasmata 16S rRNA and methyl-coenzyme M reductase (mcr) transcripts decreased concomitantly with mRNAs of enzymes involved in methanogenesis from methylamines that were among the most abundant archaeal transcripts, indicating that these Thermoplasmata degrade methylamines. Their methylotrophic methanogenic lifestyle was corroborated by in vitro incubations, showing enhanced growth of these organisms upon methylamine supplementation paralleled by elevated methane production. The Thermoplasmata have a high potential as target in future strategies to mitigate methane emissions from ruminant livestock. Our findings and the findings of others also indicate a wider distribution of methanogens than previously anticipated.

The Influence of Whole Wheat Feeding on Salmonella Infection and Gut Flora Composition in Broilers

Abstract In an experiment with broilers infected with a rifampicin-resistant Salmonella Typhimurium strain at 15 days of age, it was demonstrated that whole wheat feeding influenced the course of infection. The pH in the contents of the gizzard decreased as the amount of whole wheat in the diet increased over time. Following infection, lower numbers of Salmonella Typhimurium were found in the gizzard and ileum of birds receiving whole wheat compared to pellet-fed birds. However, there was no difference with respect to the numbers of Salmonella Typhimurium in the ceca and rectum between the two experimental groups. Quantitative cultivation of different groups of bacteria belonging to the natural intestinal microflora showed that whole wheat feeding significantly reduced the numbers of Clostridium perfringens in the intestinal tract of the birds. The present results indicate that the gizzard has an important function as a barrier organ, one that prevents pathogenic bacteria from entering the distal digestive tract.

Quantitative Determination of Bile Salt Hydrolase Activity in Bacteria Isolated from the Small Intestine of Chickens

ABSTRACT A quantitative assay based on high-performance liquid chromatography analysis of bile salts and bacterial protein determination was established for investigating bile salt hydrolase (BSH) activity in bacteria isolated from the small intestine of chickens. Bacteria were isolated using various media and were subsequently grouped according to cell morphology, fermentation profile, and 16S ribosomal DNA sequence. Representative isolates from each bacterial group were assayed for BSH activity. The isolates differed in BSH activity with respect to the state of growth and preculturing with and without taurochenodeoxycholate. The highest levels of BSH activity were found with Enterococcus faecium and Clostridium perfringens.

Influence of the oxidative quality of dietary oil on broiler meat storage stability.

Broilers were fed a high fat diet containing 11% oil (9% rapeseed oil, 2% soya bean oil) and the oil was given either as fresh (peroxide value of 1 meqv. O(2)kg(-1) oil) or as highly oxidised (peroxide value of 156 meqv. O(2)kg(-1) oil). Diets were supplemented with 46 mg all-rac-α-tocopheryl acetate kg(-1) diet, resulting in a tocopherol content of 80.8 mg α-tocopherol and 58.6 mg γ-tocopherol per kg diet in the fresh oil diet and of 44.0 mg α-tocopherol and 18.3 mg γ-tocopherol per kg diet in the oxidised oil diet, respectively, reflecting the degradation of the natural occurring tocopherols in the oxidised diet. Only minor differences were seen with respect to fatty acid composition in muscles from birds fed the two diets. The oxidation of the dietary oil lowered lipid stability significantly (p < 0.01) in both raw and precooked meats during chill storage, whereas only minor effects on the stability of frozen meat were seen. Tocopherol levels were significantly lower (p < 0.01) in muscles from birds fed the oxidised oil diet, explaining the decreased lipid stability of meat from these birds. Thigh meat was more susceptible to lipid oxidation during storage than breast meat, regardless of dietary treatment, although thigh meat had markedly higher tocopherol levels than breast meat. The molar ratio of PUFA > 18:2 (polyunsaturated fatty acids with three or more double bonds) to α-tocopherol was significantly (p < 0.01) higher in thigh meat compared with breast meat, explaining the lower stability of the former during storage.

Effect of feeding silages or carrots as supplements to laying hens on production performance, nutrient digestibility, gut structure, gut microflora and feather pecking behaviour

1. An experiment was carried out to examine the suitability of using maize silage, barley–pea silage and carrots as foraging materials for egg-laying hens. Production performance, nutrient digestibility, gastrointestinal characteristics, including the composition of the intestinal microflora as well as feather pecking behaviour were the outcome variables. 2. The protein content of the foraging material (g/kg DM) was on average 69 g in carrots, 94 g in maize silage and 125 g in barley–pea silage. The starch content was highest in the maize silage (312 g/kg DM), and the content of non-starch polysaccharides (NSP) varied from 196 to 390 g/kg, being lowest in carrots. Sugars were just traceable in the silages, whereas carrots contained on average 496 g/kg DM. 3. Egg production was highest in hens fed either carrots or maize silage, whereas hens fed barley–pea silage produced less (219 vs. 208). Although the consumption of foraging material was high (33, 35 and 48% of the total feed intake on ‘as fed’ basis for maize silage, barley–pea silage and carrots, respectively) only a minor effect on nitrogen corrected apparent metabolisable energy (AMEn) and apparent digestibility was seen. At 53 weeks of age, hens fed maize silage had AMEn and apparent digestibility values close to the control group (12·61 and 12·82, respectively), whereas access to barley–pea silage and carrots resulted in slightly lower values (12·36 and 12·42, respectively). Mortality was reduced dramatically in the three groups given supplements (0·5 to 2·5%) compared to the control group (15·2%). 4. Hens receiving silage had greater relative gizzard weights than the control or carrot-fed groups. At 53 weeks of age, the gizzard-content pH of hens receiving silage was about 0·7 to 0·9 units lower than that of the control or carrot-fed hens. Hens fed both types of silage had higher concentrations of lactic acid (15·6 vs. 3·2 µmoles/g) and acetic acid (3·6 vs. 6·1 µmoles/g) in the gizzard contents than the other two groups. The dietary supplements had a minor effect on the composition of the intestinal microflora of the hens. 5. Access to all three types of supplements decreased damaging pecking in general (to feathers as well as skin/cloaca), reduced severe feather pecking behaviour and improved the quality of the plumage at 54 weeks of age. 6. In conclusion, access to different types of foraging material such as silages and carrots improved animal welfare.

The effect of &#x1d6c3;-glucanase supplementation of barley- and oat-based diets on growth performance and fermentation in broiler chicken gastrointestinal tract

1. The aim of the present study was to investigate whether grain type (barley or oats) and β-glucanase inclusion influence the performance and the gastrointestinal ecosystem of broiler chickens, taking the concentrations of short-chain fatty acids and lactic acid, pH, ileal viscosity, and the weight of the caeca and liver into consideration. 2. The inclusion of β-glucanase in the oat-based diet improved body weight gains. Enzyme supplementation of barley-based diets improved feed conversion efficiency and reduced intestinal viscosity. 3. Irrespective of the type of cereal, β-glucanase supplementation increased the lactic acid concentration and lowered the pH of the crop contents. No such changes in fermentation were observed in the contents of the gizzard and ileum. 4. Larger amounts of total dietary fibre and its fractions (arabinoxylans and β-glucans) in oats decreased the weight of the caeca. 5. Molar ratios of acetate, propionate and butyrate in the caecal chyme were affected by cereal type but not by enzyme supplementation. The barley-based diet increased the butyrate:propionate ratio but the opposite effect was observed with the oat-based diet.

Effect of silver nanoparticles on growth performance, metabolism and microbial profile of broiler chickens

This study evaluated the potential of silver nanoparticles (AgNano) as an antimicrobial growth-promoting supplement for broiler chickens. One hundred forty-four seven-day-old broiler chicks were distributed randomly to AgNano treatments at 0, 10 and 20 mg/kg (Control, Group AgNano10, and Group AgNano20, respectively) provided via the drinking water from day 7 to 36 post-hatching. Body weight and feed consumption were measured weekly. In addition, balance and respiration experiments were carried out to determine nitrogen (N) utilisation and energy retention. At days 22 and 36, blood samples and intestinal content were collected to evaluate the effects of AgNano on plasma concentration of immunoglobulins and the intestinal microflora, repectively. The provision of water solutions containing different concentrations of AgNano had no effect on postnatal growth performance and the energy metabolism of broiler chickens. However, in Group AgNano10 N intake (p = 0.05) and retention (p = 0.03) was increased, but N excretion and efficiency of utilisation was not affected. The populations of bacteria in the intestinal samples were not affected by AgNano supplementation. The concentration of immunoglobulin (IgG) in the blood plasma of broilers supplemented with AgNano decreased at day 36 (p = 0.012). The results demonstrated that AgNano affects N utilisation and plasma IgG concentration; however, it does not influence the microbial populations in the digestive tract, the energy metabolism and growth performance of chickens.

In vitro production of necrotic enteritis toxin B, NetB, by netB-positive and netB-negative Clostridium perfringens originating from healthy and diseased broiler chickens

The Clostridium perfringens necrotic enteritis toxin B, NetB, was recently proposed as a new key virulence factor for the development of necrotic enteritis (NE) in broilers. The aim of the present study was to investigate the presence of the netB gene and the in vitro production of the NetB toxin in a well characterized collection of 48 C. perfringens Type A isolates, obtained from Danish broiler flocks. The investigation revealed netB gene prevalences of approx. 50% and 60% among isolates from diseased (NE) and healthy flocks, respectively. Only minor nucleotide variations were observed between the isolates in the coding sequence (CDS) of the netB gene, and the promoter region was observed to be completely conserved. However, in vitro NetB production was only observed in 4 out of 14 netB-positive C. perfringens isolates recovered from healthy birds, whereas 12 out of 13 netB-positive isolates from NE birds were shown to produce the NetB toxin. It is therefore proposed that genotype, i.e. presence of the netB gene, in itself is inadequate for predicting virulence of C. perfringens, and future investigations should focus on the bacterial phenotypes; the regulatory mechanisms involved in the expression of NetB, and potentially also other toxins, and its implications for the virulence of individual C. perfringens strains.