Transfer of Non-Dioxin-Like Polychlorinated Biphenyls (ndl-PCBs) from Feed and Soil into Hen Eggs.

Understanding the transfer of non-dioxin-like polychlorinated biphenyls (ndl-PCBs) into foods of animal origin is crucial for human health risk assessment. In two experiments, we investigated the transfer of ndl-PCBs from contaminated feed and soil into eggs and meat of laying hens. The transfer from the feed was investigated with 30 laying hens. The treated hens were divided into two groups fed a contaminated diet (12.8 µg/kg sum of indicator ndl-PCBs; 88% dry matter (DM)) for 28 and 63 days, respectively, and then experienced a depuration period of 100 days with control feed. The transfer from soil was investigated with 72 laying hens kept in three separate outdoor pens (with three levels of ndl-PCB soil contamination) for 168 days. In both experiments, eggs were collected and analyzed for ndl-PCBs. In the second experiment, animals (n = 3 at the beginning, n = 6 per group after 42, 84, and 168 days) were slaughtered to determine ndl-PCBs in meat (breast muscle tissue) fat. The transfer of ndl-PCB from both feed and soil was clearly measurable and concentrations in eggs quickly exceeded maximum levels. Clear differences between individual congeners were observed. In particular, the low-chlorinated ndl-PCBs 52 and 101 are hardly found in eggs, despite their relatively high concentration in feed and soil. PCBs 138, 153, and 180, on the other hand, were found in large proportions in eggs and meat.

Effect of feeding different levels of lignocellulose on performance, nutrient digestibility, excreta dry matter, and intestinal microbiota in slow growing broilers.

Lignocellulose is a constituent of plant cell walls and might be used as a fiber source in poultry nutrition. The current study investigated the impact of increasing dietary levels of lignocellulose on performance, nutrient digestibility, excreta DM, intestinal microbiota, and bacterial metabolites in slow growing broilers. At an age of 10 wk, 60 male broilers of an intercross line (New Hampshire × White Leghorn) were allocated to cages and fed isoenergetic and isonitrogenous diets containing 0.8% (LC1), 5% (LC2), or 10% (LC3) lignocellulose. After 23 D of feeding, broilers were killed and digesta samples of ileum and excreta analyzed for nutrient digestibility and DM. Cecal contents were analyzed for microbial composition and metabolites. Broiler performance was not affected by feeding dietary lignocellulose. LC3 fed broilers showed reduced ileal digestibility of protein compared to chickens fed LC1 (P = 0.003). Moreover, increasing levels of dietary lignocellulose reduced apparent digestibility of organic matter and gross energy (P < 0.001). Feeding of lignocellulose had no impact on the excreta DM of broilers. Increasing levels of dietary lignocellulose lowered cecal counts of Escherichia/Hafnia/Shigella (P = 0.029) and reduced the total concentration of short-chain fatty acids (P < 0.001), lactate (P < 0.05), and ammonia (P = 0.009). The molar ratio of cecal acetic acid was higher in LC3 fed broilers (P < 0.001), while the proportions of cecal propionic acid and n-butyric acid were higher in LC1 and LC2 fed chickens (P < 0.001). Correlation analyses indicated that dietary lignocellulose was negatively related to the total concentration of cecal bacterial metabolites (P < 0.001). In conclusion, the feeding of lignocellulose did not affect growth performance, but impaired nutrient digestibility of slow growing broilers. While minor changes in cecal microbial composition were detected, cecal bacterial metabolite concentrations were significantly reduced with increasing levels of dietary lignocellulose. These findings suggest that lignocellulose is not extensively degraded by bacteria residing in the large intestine of broilers.

Effect of a "diluted" diet containing 10% lignocellulose on the gastrointestinal tract, intestinal microbiota, and excreta characteristics of dual purpose laying hens.

Low performing dual purpose hens have different nutritional requirements compared to conventional hybrid hens. Lignocellulose is a low fermentable polymer, acting as a diet diluent and may influence physiological and digestive processes. This study investigated the effect of a 10% dietary lignocellulose dilution on the development of gastrointestinal organs, intestinal morphology, intestinal microbiota, and excreta characteristics of dual purpose hens. One-day-old female Lohmann Dual chicks were allocated to 12 pens and fed two different diets: A standard control diet (CON) and a treatment diet (LC), based on CON but diluted with 10% lignocellulose (ARBOCEL®). At 52 wk of age, gastrointestinal organs were extracted and weights determined. Colorectal tissue samples were chemically fixed and stained for histomorphological examinations. Cecal digesta samples were analyzed for bacterial metabolites and composition using gas chromatography, HPLC, photometry, and PCR. Excreta dry matter and viscosity was consistently assessed during the trial. LC-fed hens showed increased weights of the gizzard (P = 0.003), small (P < 0.001), and large intestine (P = 0.048) compared to hens fed CON. LC-fed hens had a larger colorectal villus area (P = 0.049), a higher mucosal enlargement factor of villi (P = 0.016) and crypts (P = 0.030) than CON-fed hens. The concentration of short-chain fatty acids (SCFAs) (P = 0.017) and ammonia (P = 0.013) was higher in CON-fed hens compared to LC-fed hens. Bacterial composition and activity was generally not affected by feeding the different diets. LC-fed hens had a higher excreta dry matter content than hens fed CON at 10 (P < 0.001), 17 (P < 0.001), and 22 (P = 0.002) wk of age. Correlation analyses revealed a negative relationship between the concentration of SCFAs in the cecum and the colorectal villus surface area (P < 0.01). In conclusion, the feeding of high levels of lignocellulose increased gastrointestinal organ weights and colorectal surface area in dual purpose laying hens. A higher intestinal surface area in combination with lower concentrations of SCFAs might indicate a compensatory reaction of hens fed LC enhancing the absorption of bacterial metabolites by increasing the intestinal mucosal surface.

Influence of differently processed yeast (Kluyveromyces fragilis) on feed intake and gut physiology in weaned pigs.

Two feeding trials were conducted to investigate the effects of hydrolyzed (HY) or non-hydrolyzed (NHY) yeast (Kluyveromyces fragilis) in isoenergetic and isonitrogeneous diets in the postweaning period. In experiment 1, a total of 550 unsexed pigs (6.5 ± 0.5 kg BW), weaned at 24 ± 2 d of age, were allocated to five treatment groups, receiving either a control diet (CON) or diets with 1%, 3%, and 5% HY (groups HY1, HY3, and HY5, respectively), or a diet with 3% NHY (group NHY3). In experiment 2, a total of 48 male and female pigs (6.2 ± 0.3 kg BW, weaned at d 25) were allocated to three dietary groups (n = 8 replicates with two pigs) receiving a control diet (CON) or diets with 1% NHY or 1% HY. Eight animals were sacrificed 2 wk after weaning for histological investigations in the jejunum and colon, determination of apparent ileal digestibility (AID) of CP and ether extract (EE), and electrophysiological measurements in the jejunal tissue after addition of carbachol or l-glutamine using Ussing chambers. In experiment 1, different treatments had no significant effect on pig performance, but diet HY1 tended to increase ADG and G:F in wk 2 after weaning (P < 0.1). In experiment 2, diet HY1 increased feed intake in wk 2 (P < 0.05), whereas NHY yeast had no effect on feed intake. Villus height, villus/crypt ratio in jejunum (P < 0.05), and crypt depth in colon (P < 0.01) were increased in group HY1. Crypt depth in jejunum and small intestinal length were not affected by different treatments. The AID of CP and EE tended to increase in group HY1 (P < 0.1) compared with groups CON and NHY. In the Ussing chamber experiments, no changes in basal electrophysiological parameters were observed, and the reactions of the treatment groups to carbachol and l-glutamine were comparable. ADFI was positively correlated with different parameters of intestinal morphology (villus height, villus/crypt ratio, crypt depth in colon, length of small intestine), AID of CP, EE, and performance. The results suggest that a supplementation of 1% HY based on K. fragilis to pig diets may positively influence ADFI and intestinal morphology in pig in the early postweaning period (d 1 to 14).

Effect of feeding soybean meal and differently processed peas on intestinal morphology and functional glucose transport in the small intestine of broilers.

Peas are locally grown legumes being rich in protein and starch. However, the broad usage of peas as a feed component in poultry nutrition is limited to anti-nutritional factors, which might impair gut morphology and function. This study investigated the effect of feeding raw or differently processed peas compared with feeding a soybean meal-based control diet (C) on intestinal morphology and nutrient transport in broilers. A total of 360 day-old broiler chicks were fed with one of the following diets: The C diet, and 3 diets containing raw peas (RP), fermented peas (FP) and enzymatically pre-digested peas (EP), each supplying 30% of dietary crude protein. After 35 d, jejunal samples of broilers were taken for analyzing histomorphological parameters, active glucose transport in Ussing chambers and the expression of genes related to glucose absorption, intestinal permeability and cell maturation. Villus length (P = 0.017) and crypt depth (P = 0.009) of EP-fed broilers were shorter compared to birds received C. The villus surface area was larger in broilers fed C compared to those fed with the pea-containing feed (P = 0.005). Glucose transport was higher for broilers fed C in comparison to birds fed with the EP diet (P = 0.044). The sodium-dependent glucose co-transporter 1 (SGLT-1) expression was down-regulated in RP (P = 0.028) and FP (P = 0.015) fed broilers. Correlation analyses show that jejunal villus length negatively correlates with the previously published number of jejunal intraepithelial T cells (P = 0.014) and that jejunal glucose transport was negatively correlated with the occurrence of jejunal intraepithelial leukocytes (P = 0.041). To conclude, the feeding of raw and processed pea containing diets compared to a soybean based diet reduced the jejunal mucosal surface area of broilers, which on average was accompanied by lower glucose transport capacities. These morphological and functional alterations were associated with observed mucosal immune reactions. Further studies are required elucidating the specific components in peas provoking such effects and whether these effects have a beneficial or detrimental impact on gut function and animal health.

Efficacy of a single oral dose of a live bivalent E. coli vaccine against post-weaning diarrhea due to F4 and F18-positive enterotoxigenic E. coli.

F4- and F18-positive enterotoxigenic E. coli strains (F4-ETEC and F18-ETEC) are important causes of post-weaning diarrhea (PWD) in pigs. F4 (antigenic variant ac) and F18 (ab and ac) fimbriae are major antigens that play an important role in the early stages of infection. Herein, the efficacy of a live oral vaccine consisting of two non-pathogenic E. coli strains, one F4ac- and one F18ac-positive, was evaluated using F4ac-ETEC and F18ab-ETEC challenge models. A randomized, masked, placebo-controlled, block design, parallel-group confirmatory study with two different vaccination-challenge intervals (7 and 21 days) was conducted for each challenge model. The vaccine was administered in one dose, to ≥18-day-old piglets via drinking water. Efficacy was assessed by evaluating diarrhea, clinical observations, weight gain and fecal shedding of F4-ETEC or F18-ETEC. Anti-F4 and anti-F18 immunoglobulins in blood were measured. The vaccination resulted in significant reductions in clinical PWD and fecal shedding of F4-ETEC and F18-ETEC after the 7- and 21-day-post-vaccination heterologous challenges, except for after the 21-day-post-vaccination F4-ETEC challenge, when the clinical PWD was too mild to demonstrate efficacy. A significant reduction of mortality and weight loss by vaccination were observed following the F18-ETEC challenge. The 7-day protection was associated with induction of anti-F4 and anti-F18 IgM, whereas the 21-day protection was mainly associated with anti-F4 and anti-F18 IgA. The 7-day onset and 21-day duration of protection induced by this vaccine administered once in drinking water to pigs of at least 18days of age were confirmed by protection against F4-ETEC and F18-ETEC, and induction of F4 and F18-specific immunity. Cross protection of the vaccine against F18ab-E. coli was demonstrated for both the 7- and 21-day F18-ETEC challenges.

Effect of feeding soybean meal and differently processed peas on the gut mucosal immune system of broilers.

Peas are traditionally used as a protein source for poultry. However, peas contain antinutritional factors (ANF), which are associated with the initiation of local and systemic immune reactions. The current study examined the effect of feeding raw or differently processed peas in comparison with feeding a soybean meal (SBM) based control diet (C) on the gut mucosal immune system of broilers in a 35 day feeding trial. In six replicates, a total of 360 one-day-old male broilers were randomly allocated to four different groups receiving C, or three treatment diets containing raw, fermented, and enzymatically pre-digested peas, each supplying 30% of required crude protein. After slaughtering, jejunal samples were taken for immunohistochemical, flow cytometric, and gene expression analyses. Investigations were focused on the topological distribution of intraepithelial leukocytes (villus tip, villus mid, and crypt region) as well as on the further characterization of the different intraepithelial lymphocytes (IEL) and concomitant pro- and anti-inflammatory cytokines. Broilers receiving the raw or processed pea diets had higher numbers of intraepithelial CD45+ leukocytes in the tip (P = 0.004) and mid region (P < 0.001) of villi than birds fed C. Higher numbers of intraepithelial CD3+ lymphocytes were found in the villus tip (P = 0.002) and mid region (P = 0.003) of birds fed raw or processed pea containing diets in comparison with those fed C. The flow cytometric phenotyping showed a similar relative distribution of IEL among the feeding groups. The expression of intestinal pro- and anti-inflammatory cytokines was affected by feeding the different diets only to a minor extent. To conclude, feeding of diets formulated with raw and processed peas in comparison with feeding a SBM control diet initiated mucosal immune responses in the jejunum of broilers indicated by a quantitative increase of intraepithelial T cells. Further research is needed in order to ascertain the specific factors which are responsible for observed local immune reactions and how these local reactions might affect the immune status and health of broilers.

Bovine milk-based formula leads to early maturation-like morphological, immunological, and functional changes in the jejunum of neonatal piglets.

Artificial rearing and formula feeding is coming more into the focus due to increasing litter sizes and limited nursing capacity of sows. The formula composition is important to effectively support the development of the gut and prevent intestinal dysfunction in neonatal piglets. In this study, newborn piglets ( = 8 per group) were fed a bovine milk-based formula (FO), containing skimmed milk and whey as the sole protein and carbohydrate sources, or were suckled by the sow (sow milk [SM]). After 2 wk, tissue from the jejunum was analyzed for structural (i.e., morphometry) and functional (i.e., disaccharidase activity, glucose transport, permeability toward macromolecules, and immune cell presence) changes and concomitant expression of related genes. Formula-fed piglets had more liquid feces ( < 0.05) over the entire experimental period. Although FO contained twice as much lactose (46% on a DM basis) as SM (21%) and no maltose or starch, the lactase activity was lower ( < 0.05) and glucose transport capacity was higher ( < 0.05) in FO-fed pigs. The relative proportion of intraepithelial natural killer cells and proinflammatory cytokine gene expression (, , and ) was higher in FO-fed pigs ( < 0.05). Piglets fed FO had deeper crypts, larger villus area, and higher expression of caspase 3 and proliferating cell nuclear antigen ( < 0.05). Epithelial permeability toward fluorescein isothiocyanate-dextran was higher and expression of claudin-4 was lower in FO-fed piglets ( < 0.05). The data suggest an early response to bovine milk-based compounds in the FO accompanied with early onset of functional maturation and impaired barrier function. Whether lactose, absence of species-specific protective factors, or antigenicity of foreign proteins lead to to the observed intestinal reactions requires further clarification.

Feed supplemented with organic acids does not affect starch digestibility, nor intestinal absorptive or secretory function in broiler chickens.

The current study aimed to determine the impact of acidified feed on apparent ileal starch digestibility, intestinal transport and barrier function and intestinal glucose transporter expression. The experiment included a control group and a treatment group with broilers fed a standard diet without or with 1.5% of a commercial organic acid product (64% formic acid, 25% propionic acid, 11% water). Broilers were fed with the experimental diets from hatching until days 32-35. Starch digestibility was determined using 0.2% titanium dioxide as ingestible marker. Gene expressions of the intestinal sodium glucose transporter 1 (SGLT-1) and glucose transporter 2 (GLUT-2) were analysed using qPCR analysis. Additionally, SGLT-1 function and chloride secretion were analysed in Ussing chamber experiments. Jejunal samples were sequentially exposed to 10 mm glucose, 100 µm phloridzin, 100 µm histamine and 100 µm carbachol. Apparent ileal starch digestibility (±SEM) of the control group (97.5 ± 0.35%) and the acid-treated group (97.0 ± 0.59%) did not differ (p = 0.674). The mean tissue conductance of intestinal samples obtained from the control group and the treatment group was similar [10.6 mS/cm(2) (±0.68) and 9.4 mS/cm(2) (±0.80) respectively (p = 0.147)]. The mean short-circuit currents (ΔIsc ) of the samples exposed to glucose, phloridzin, histamine and carbachol did not differ (p > 0.05). Additionally, no differences in the expression of SGLT-1 and GLUT-2 could be observed (p = 0.942, p = 0.413). Based on this study, the consumption of feed supplemented with organic acids was not associated with effects on ileal starch digestibility and functional traits of jejunal tissues, indicating that these additives have no major impact on the small intestinal function in broilers.

Implication of milling methods, thermal treatment, and particle size of feed in layers on mineral digestibility and retention of minerals in egg contents.

Feed production with different milling methods, thermal treatment, and particle size may influence mineral digestibility and retention in eggs. The present study investigated the impact of roller (R) and hammer (H) mills, mash (M) and expandate (E) with fine (F) and coarse (C) particle sizes, on apparent ileal absorption (AIA) and apparent total digestibility (ATD) and retention of calcium, phosphorus, magnesium, zinc, manganese, copper and iron in yolk, albumen, and shell. A total of 384 hens (Lohmann Brown), 19 weeks old, were assigned using a randomized design with a 2×2×2 factorial arrangement. Eight experimental diets were offered ad libitum during the whole experimental period and one week before for diet adaption. The AIA of magnesium, zinc, copper, and iron was higher in treatment R in comparison with treatment H (P<0.01, P≤0.03, P<0.01 and P<0.01, respectively). The AIA of magnesium was higher in treatment M than treatment E (P<0.01). The AIA of magnesium was higher in treatment C in comparison with treatment F (P≤0.05) due to particle size. The ATD of copper and iron was higher in treatment R than treatment H (P<0.01 and P≤0.03, respectively). The ATD was higher for phosphorus and lower for iron in treatment F than treatment C (P≤0.05 and P≤0.02. respectively). The copper concentration in yolk and albumen was higher in treatment C than treatment F (P<0.01 and P≤0.03, respectively). Besides a few overall interactions, the AIA and ATD of copper and manganese were lower in H+M group than R+M group (P≤0.05). The ATD of iron was higher in the M+C group compared to the M+F group (P<0.01), whereas the albumen zinc concentration was higher in the E+C group than E+F group (P < 0.01). In conclusion, the feed produced by hammer mill had negative effects on AIA and ATD for trace elements in particular, but mineral concentrations in egg contents were mostly comparable for all treatments. Therefore, milling methods, thermal treatment, and particle sizes used in the present study can be used for layer feed formulation without negatively affecting egg quality.

The effects of different thermal treatments and organic acid levels in feed on microbial composition and activity in gastrointestinal tract of broilers.

Thermal treatments of feed and organic acids are known to affect the gastrointestinal microbiota in chickens. The present study evaluated the effect of different thermal processes including pelleting (P), long-term conditioning at 85°C for 3 min (L), expanding at 110°C (E110), and 130°C for 3 to 5 s (E130) as well as organic acid (63.75% formic acid, 25.00% propionic acid, and 11.25% water) inclusion levels (0, 0.75, and 1.5%) on gastrointestinal microbiota in broilers. In total, 960 one-day-old chicks were randomly assigned to 8 replicates using a 3 × 4 factorial arrangement. At d 35, bacterial cell numbers in the crop, ileum, and cecum, and bacterial metabolites in the crop, gizzard, ileum, and cecum were determined. The inclusion of 1.5% organic acids increased cell numbers of all clostridial clusters in the crop. The organic acid supplementation increased the propionic acid concentration in the crop and gizzard and there was a decrease in lactic acid concentration. In the ileum, the 0% organic acid group had the highest numbers of Lactobacillus spp. and enterobacteria. Inclusion of 1.5% organic acids increased ileal acetate concentration. Increasing the feed processing temperature led to an increase of lactobacilli in the crop and ileum, whereas clostridia and enterobacteria seemed unaffected. Similarly, lactate concentrations increased in the ileum, but short-chain fatty acids remained identical. In the crop, an increase for acetate was found for the E130 group compared with all other thermal treatments. In conclusion, our study demonstrated that thermal treatments and organic acid supplementation to broiler diets more markedly influenced the bacterial status of the crop compared with the downstream segments and their effects decreased along the length of gastrointestinal tract. Whereas organic acids markedly modified bacterial composition and activity in the crop, expansion increased lactobacilli and lactate in the crop and ileum.

Effects of grinding method, particle size, and physical form of the diet on gastrointestinal morphology and jejunal glucose transport in laying hens.

Several studies illustrated that the structure of feed, i.e., the particle size, particle-size distribution, and the physical form of the diet, affects the avian gastrointestinal function and health leading to changes in productive performance. However, investigations concerning the effects of feeding differently processed diets on laying hens are limited and primarily concentrated on bird performance. The current study examines the effect of feed processing on the gastrointestinal morphology and on the jejunal glucose transport of laying hens. In 8 replicates, a total of 384 hens (Lohmann Brown) aged 20 wk were randomly allocated to 8 different groups and fed over a period of 21 d in a 3-factorial design. Diets differed in 1) grinding method, either hammer or roller mill; 2) physical form, either mash or expandate; and 3) particle size, either coarsely or finely ground. During the experimental trial, the laying performance of each feeding group was recorded daily and the feed intake and BW determined weekly. After slaughtering, the weights of the pancreas, proventriculus, gizzard, and small intestine were measured. Villus lengths and crypt depths of the duodenum, jejunum, and ileum were determined. The jejunal electrogenic glucose transport was studied in Ussing chambers. Hens that received mash instead of expandate had higher proventriculus (P = 0.011), gizzard (P < 0.001), and pancreas (P = 0.019) weights, whereas the feeding of coarsely instead of finely ground diets led to higher gizzard weights (P < 0.001). Mash-fed hens showed longer duodenal (P < 0.001) and shorter ileal villi (P = 0.047) and increased duodenal villus height-to-crypt depth ratios (P < 0.001) than those given the expandate. Mash-fed hens had higher glucose transport rates than expandate-fed hens (P < 0.001). In conclusion, the feeding of coarsely ground as well as mash diets had stimulating effects on the development of the gastrointestinal organs. Moreover, the feeding of mash influenced the intestinal microstructure of the epithelium that was accompanied by higher glucose transport capacities.

The effects of different thermal treatments and organic acid levels on nutrient digestibility in broilers.

Poultry feed is a potential vector for pathogens. Heat processing and organic acid treatments may decontaminate feed and can affect bird performance as well as feed digestibility. The present study was performed to investigate the effect of different thermal treatments including pelleting (P), long-term conditioning at 85°C for 3 min (L), or expanding at 110°C (E110) and 130°C for 3 to 5 s (E130) without or with 0.75 and 1.5% organic acid supplementation (63.75% formic acid, 25.00% propionic acid, and 11.25% water) on performance, nutrient digestibility, and organ weights of broilers. In total, 960 one-day-old broiler chicks were randomly assigned to 8 replicates using a 3 × 4 factorial arrangement. Performance variables were determined, and the relative organ weights and ileal and total amino acid (AA) digestibilities were measured at d 35. The organic acid inclusion linearly improved feed efficiency in the first week (P ≤ 0.05). The acid inclusion levels and thermal treatments had no significant effect on the performance variables at later intervals of the growing period of the birds. The L group showed the lowest ileal AA and CP digestibility. The inclusion of organic acids had a quadratic effect on total and ileal digestibility of isoleucine (P ≤ 0.05), whereas it had no significant effect on the ileal digestibility of other AA and nutrients. The relative weights of the jejunum and small intestine were significantly higher in the E130 group compared with P and L (P ≤ 0.05). In conclusion, our study demonstrated that long-term heat conditioning can decrease ileal nutrient digestibility, whereas pelleting and expansion, independently of organic acid addition, seemed to have no negative impact on broiler performance and nutrient digestibilities. Moreover, adding a blend of organic acids to broiler diets had neither positive nor negative effects on nutrient digestibility and final broiler performance. This indicates the feasibility of short-term thermal treatment and acid supplementation for hygienization of broiler feed without negatively influencing performance.

Impact of thermal and organic acid treatment of feed on apparent ileal mineral absorption, tibial and liver mineral concentration, and tibia quality in broilers.

Minerals play an important role for growth and bone stability in broilers. Thermal treatment and inclusion of organic acids in feed may affect the mineral absorption and tibial quality in broilers. The study was conducted to investigate the effect of thermal processing of feed including pelleting (P), long-term conditioning at 85°C (L), and expanding at 130°C (E) without and with 1.5% of an acid mixture containing 64% formic and 25% propionic acid on the apparent ileal absorption (AIA) of calcium, phosphorus, magnesium, potassium, sodium, iron, copper, manganese, and zinc, their concentrations in liver and tibia, as well as various tibial quality parameters in broilers. In total, 480 one-day-old Cobb broiler chicks were assigned using a completely randomized design with a 3 × 2 factorial arrangement. The ileal digesta, liver, and tibia were collected at d 35. The AIA of calcium and sodium was improved in group E compared with L (P ≤ 0.02 and P ≤ 0.01). Group P and E showed higher AIA for potassium than L (P ≤ 0.01). Bone ash content was increased in group E compared with L (P ≤ 0.04). The BW to bone weight ratio was lower and tibial zinc content was higher in group P compared with E (P ≤ 0.05). Tibial iron content was higher in group L than E (P ≤ 0.03). Acid addition did not affect AIA, mineral content in tibia, or tibial quality parameters. Thermal and acid treatment did not affect mineral concentrations in the liver, except an inconsistent interaction effect for DM content and sodium (P ≤ 0.03 and P ≤ 0.04, respectively). In conclusion, long-term thermal treatment reduced AIA of some minerals compared with short-term thermal treatments, but had no impact on tibia composition. Acid inclusion had no effect on AIA of minerals and tibia quality. Thermal treatment and the use of organic acids can therefore be considered as safe with regard to their impact on bone development in broilers.