Effects of different dietary threonine and glycine supplies in broilers fed low-protein diets.

The reduction of crude protein (CP) content of broiler diets with balanced amino acid supply can increase the nitrogen (N) utilization efficiency and reduce ammonia emission, the risk of many health problems in birds. Feeding low protein (LP) diets without the impairment of performance traits needs the optimized dietary levels of threonine (Thr) and the non-essential amino acid (AA) glycine (Gly) and serine (Ser). However, the required concentrations and interactions of Thr and Gly + Ser, expressed as Gly equivalent (Glyequi), in LP diets are not fully understood. Therefore, the aim of this study was to investigate the effects of three LP (LP1-3) grower (11-24 days) and finisher (25-35 days) diets with 2% CP reduction compared to the control (C), differing in standardized ileal digestible (SID) Thr to lysine (Lys) ratio (C, LP1, LP3: 63%, LP2: 72%) and Glyequi levels (C: 15.65 g/kg, LP1: 13.74 g/kg, LP2: 13.70 g/kg, LP3: 15.77). The LP treatments did not impair the performance traits of broilers. The LP2 treatment with increased SID Thr-to-Lys ratio (+9.0%) resulted in significantly higher body weight gain and a more advantageous feed conversion ratio in the whole fattening compared to the control treatment with normal CP level (p < 0.05). The LP3 treatment containing swine meat meal with similar Glyequi levels compared to the normal CP treatment led to the most advantageous feed conversion ratio in the finisher phase and the highest nitrogen retention efficiency (p < 0.05). However, the LP3 treatment with a high starch-to-CP ratio negatively influenced the relative carcass weight and the ratio of abdominal fat of broilers (p < 0.05).

Near-Infrared (NIR) Spectroscopy as an Alternative for Predicting n-Alkane Concentration in Excreta of Laying Hens: NIR-Generated Data for Dietary Composition Estimation.

N-alkanes offer a promising approach for assessing the nutritional contribution of external sources to the diets of laying hens in free-range production systems. However, traditional laboratory methods, involving extraction, purification and gas chromatographic analysis, are both economically burdensome and time-consuming. Near-infrared spectroscopy (NIRS) is emerging as a viable alternative, with varying degrees of accuracy depending on the chemical nature and concentration of the component of interest. In our research, we focus on the accuracy of NIRS in predicting the concentrations of n-alkanes (C25-C33) in excreta under simulated free-range conditions with two different diets: one containing a commercial feed with minimal n-alkane content and another containing 1% alfalfa on top of the commercial feed. Spectra processing and calibration were tailored for each n-alkane, with NIRS performance influenced by diet type. Notably, plant predictions using NIR-generated data were consistent with laboratory results, despite a slight tendency toward overestimation (3.40% using the NIRS-generated C25-C29-C33 combination versus 2.80% using laboratory analysis). This indicates the potential of NIRS as an efficient tool to assess n-alkanes in excreta of laying hens and, consequently, the nutritional contribution of the free-range environment, providing rapid and cost-effective results.

Feeding sunflower meal with pullets and laying hens even at a 30% inclusion rate does not impair the ileal digestibility of most amino acids.

The use of locally available protein sources in poultry nutrition is challenging for feed manufacturers and farmers. Sunflower meal (SFM) is available in high quantities in several European countries and could be used as a poultry feedstuff at higher inclusion rates. However, its maximum inclusion rate in the diets of different poultry species and age categories is unknown. Pullets and laying hens can probably tolerate higher amounts of SFM, but only limited information is available on these poultry groups. Therefore, a digestibility trial was carried out with 8-week-old layer type pullets and 50-week-old laying hens. Beside a basal diet, SFM was fed at 10, 20 and 30% inclusion rates. Feeding SFM significantly improved the digestibility of essential amino acids (AA) of threonine, valine, lysine, tyrosine, glycine, aspartic acid, and arginine in the pullet diets. No such improvement was found in laying hens. Only the absorption of the two branch-chain AAs, leucin (pullets) and isoleucine (hens), declined due to SFM. The AA digestibility of the SFM itself was also calculated by linear regression. The coefficients were, in all cases, higher in hens than in pullets. Comparing the measured digestibility coefficients of SFM with table values, it can be concluded that high variance exists because of the differences in the methodology and the test animals in the digestibility trials. From the present trial, it can be concluded that SFM can entirely replace extracted soybean meal in pullet and layer diets, without negative effects on the protein digestion of birds.

Quantitative and Qualitative Evaluation of Plant Intake in Laying Hens: n-Alkanes as Predictive Fecal Markers for Dietary Composition Assessment.

The shift in animal welfare standards towards free-range housing for laying hens in the EU has raised questions about changes in dietary composition. Accurate assessment of outdoor plant material intake is crucial for effective feeding strategies. This study introduces an approach using n-alkanes as markers to determine plant intake in laying hens, involving n-alkane recovery rate assessment, discriminant analysis and linear equation-solving for both qualitative and quantitative assessment, respectively, considering systematic n-alkane combinations. Two diets: a standard commercial diet and a diet incorporating 1% alfalfa were tested. Chemical analyses showed an altered n-alkane profile due to alfalfa inclusion, resulting a recovery rates ranging from 30-44% depending on the n-alkane type and diet. Statistical analysis revealed significant differences in recovery rates among the different alkanes for the same diets and between the diets for the same alkane, together with an interaction between n-alkane carbon chain length and initial concentration in the diet. The method accurately predicted plant inclusion, with a slight overestimation (2.80%) using the combination C25-C29-C33. Accurate qualitative classification of the animals based on fecal n-alkanes profiles was observed. The study successfully demonstrated the utility of n-alkanes for estimating dietary composition, providing a non-invasive approach for future free-range studies.

The Hatching Time of Broiler Chickens Modifies Not Only the Production Traits but Also the Early Bacteriota Development of the Ceca.

This trial was carried out to find out the effects of the parent flock and hatching time of broiler chickens on the production traits and bacteriota development of animals. Two sets of 730 hatching eggs were collected from two different parent flocks with ages of 25 and 50 weeks. In the hatchery, both groups were divided into two subgroups: those hatched during the first 10 and the subsequent 10 h of the hatching window. A feeding trial was carried out afterwards, using the four treatments in six replicate floor pens and feeding commercial starter, grower, and finisher diets that contained all the nutrients according to the breeder's recommendations. The day-old chickens of the older parent flock and those hatched later were heavier, and this advantage remained until the end of the production period. The different ages and origins of the parent flocks failed to modify the microbiological parameters of the chicken's ceca; however, the hatching time significantly influenced the different bacteriota diversity indices: the late-hatched chickens showed higher Bacteroidetes and lower Firmicutes and Actinobacteria abundances at day 11. These treatments resulted in differences in the main families, Ruminococcaceae, Lactobacillaceae, and Bacteroidaceae. These differences could not be found at day 39.

Antimicrobial Peptides (AMP) in the Cell-Free Culture Media of Xenorhabdus budapestensis and X. szentirmaii Exert Anti-Protist Activity against Eukaryotic Vertebrate Pathogens including Histomonas meleagridis and Leishmania donovani Species.

Anti-microbial peptides provide a powerful toolkit for combating multidrug resistance. Combating eukaryotic pathogens is complicated because the intracellular drug targets in the eukaryotic pathogen are frequently homologs of cellular structures of vital importance in the host organism. The entomopathogenic bacteria (EPB), symbionts of entomopathogenic-nematode species, release a series of non-ribosomal templated anti-microbial peptides. Some may be potential drug candidates. The ability of an entomopathogenic-nematode/entomopathogenic bacterium symbiotic complex to survive in a given polyxenic milieu is a coevolutionary product. This explains that those gene complexes that are responsible for the biosynthesis of different non-ribosomal templated anti-microbial protective peptides (including those that are potently capable of inactivating the protist mammalian pathogen Leishmania donovanii and the gallinaceous bird pathogen Histomonas meleagridis) are co-regulated. Our approach is based on comparative anti-microbial bioassays of the culture media of the wild-type and regulatory mutant strains. We concluded that Xenorhabdus budapestensis and X. szentirmaii are excellent sources of non-ribosomal templated anti-microbial peptides that are efficient antagonists of the mentioned pathogens. Data on selective cytotoxicity of different cell-free culture media encourage us to forecast that the recently discovered "easy-PACId" research strategy is suitable for constructing entomopathogenic-bacterium (EPB) strains producing and releasing single, harmless, non-ribosomal templated anti-microbial peptides with considerable drug, (probiotic)-candidate potential.

Representative Survey for Evaluating Housing and Manure Handling Technologies of the Hungarian Pig Sector.

In Hungary, there is a lack of information on the pig production technologies in place in the base year of 2005 and changes since then, as well as a lack of information on the number of pigs kept in different age and production categories, which makes it difficult to calculate ammonia emissions and reductions in the national inventories. Our research team conducted a representative survey of pig farms to assess housing and manure management technologies in the Hungarian pig sector in 2005 and 2015. Novel expert-based calculation methods were developed to convert farm data on pig populations into daily average numbers (DAN) of animals in different statistical categories and feeding phases. The survey resulted in a representative database of housing, manure handling, storage and manure application practices in Hungarian pig production. The data and methodology from the survey helped to develop an ammonia emission calculator and knowledge transfer tool (AGEM-S) for use by farmers.

Effects of Feeding Low Protein Diets with Different Energy-to-Protein Ratios on Performance, Carcass Characteristics, and Nitrogen Excretion of Broilers.

This study shows the effects of feeding low protein (LP) diets with different energy-to-protein ratios were evaluated on the production parameters, carcass composition, meat quality, nitrogen retention, and excreta composition of broilers. A total of 576-day-old Ross 308 broilers were fed a control diet (C) and three LP diets containing 1.5% less crude protein than diet C for 41 days. The LP1 treatment was isocaloric with diet C, while the dietary apparent metabolizable energy corrected by nitrogen (AMEn) levels in the case of the LP2 and LP3 treatments were reduced by 1.5% and 3%, respectively. The LP diets were supplemented with six crystalline essential amino acids (AA) to meet the standardized ileal digestible AA requirements of broilers. The LP1 treatment did not affect the performance parameters of broilers and increased the breast meat yield, the nitrogen retention and decreased drip loss of breast meat and the total-N and uric acid-N nitrogen excretion of birds in comparison with the C group. Although the energy-reduced LP2 and LP3 diets resulted in lower final body weight, they did not affect the carcass composition, breast meat quality, nitrogen retention, and excreta composition of birds compared with the control treatment.

XENOFOOD-An Autoclaved Feed Supplement Containing Autoclavable Antimicrobial Peptides-Exerts Anticoccidial GI Activity, and Causes Bursa Enlargement, but Has No Detectable Harmful Effects in Broiler Cockerels despite In Vitro Detectable Cytotoxicity on LHM Cells.

Entomopathogenic bacteria are obligate symbionts of entomopathogenic nematode (EPN) species. These bacteria biosynthesize and release non-ribosomal-templated hybrid peptides (NR-AMPs), with strong, and large-spectral antimicrobial potential, capable of inactivating pathogens belonging to different prokaryote, and eukaryote taxa. The cell-free conditioned culture media (CFCM) of Xenorhabdus budapestensis and X. szentirmaii efficiently inactivate poultry pathogens like Clostridium, Histomonas, and Eimeria. To learn whether a bio-preparation containing antimicrobial peptides of Xenorhabdus origin with accompanying (in vitro detectable) cytotoxic effects could be considered a safely applicable preventive feed supplement, we conducted a 42-day feeding experiment on freshly hatched broiler cockerels. XENOFOOD (containing autoclaved X. budapestensis, and X. szentirmaii cultures developed on chicken food) were consumed by the birds. The XENOFOOD exerted detectable gastrointestinal (GI) activity (reducing the numbers of the colony-forming Clostridium perfringens units in the lower jejunum. No animal was lost in the experiment. Neither the body weight, growth rate, feed-conversion ratio, nor organ-weight data differed between the control (C) and treated (T) groups, indicating that the XENOFOOD diet did not result in any detectable adverse effects. We suppose that the parameters indicating a moderate enlargement of bursas of Fabricius (average weight, size, and individual bursa/spleen weight-ratios) in the XENOFOOD-fed group must be an indirect indication that the bursa-controlled humoral immune system neutralized the cytotoxic ingredients of the XENOFOOD in the blood, not allowing to reach their critical cytotoxic concentration in the sensitive tissues.

Diet Composition and Using Probiotics or Symbiotics Can Modify the Urinary and Faecal Nitrogen Ratio of Broiler Chicken's Excreta and Also the Dynamics of In Vitro Ammonia Emission.

The objective of this research was to determine whether diet composition, or adding probiotic or symbiotic feed additives to broiler diets can modify the N composition of the excreta and the dynamics of ammonia volatilization from the manure. A total of 574 one-day-old Ross 308 broiler chickens were fed four different diets. The treatments included a corn and soybean meal-based control diets (C), wheat-based and wheat bran containing diets (W), a multi-strain probiotic treatment (Broilact®; Br), and a symbiotic additive containing Bacillus subtilis, inulin, and Saccharomices cerevisiae (Sy). Feeding the wheat-based diet significantly improved the weight gain and FCR of chickens. Treatment W also significantly increased the dry matter content of the excreta compared with the probiotic and symbiotic treatments. Both Br and Sy tended to decrease the amount of excreted uric acid, which is the main substrate of ammonia. Treatment Sy reduced the urinary N ratio of the excreta in comparison with treatment W. The symbiotic additive resulted in significantly higher ammonia emission in the first two hours. On the other hand, the dynamics of the emission was slow at the beginning and increased steeply after 15 h when the wheat-based diets were fed. Based on our results, the wheat-based diets, containing soluble arabinoxylans, and the symbiotic treatments of broiler diets have an impact on the urinary and faecal nitrogen composition of the excreta, and also on the dynamics of ammonia release from the manure.

Effects of Dietary Milk Thistle (Silybum marianum) Supplementation in Ducks Fed Mycotoxin-Contaminated Diets.

The medicinal plant milk thistle (Silybum marianum) has been widely used due to its hepatoprotective properties. The main objective of our study was to investigate the health protective effects of dietary milk thistle seed (MS), oil (MO), and seed cake (MSC) in ducks fed diets naturally contaminated with deoxynivalenol (DON; 3.43-3.72 mg/kg feed) and zearalenone (ZEN; 0.46-0.50 mg/kg feed). Female White Hungarian ducks were randomly allocated to four dietary treatments consisting of the control diet (C), the control diet supplemented with 0.5% MS, 0.5% MSC, or 0.1% MO. The feeding of experimental diets did not result in mortality cases, clinical signs of mycotoxicosis, or in differences of clinical chemistry values of blood serum. The positive effect of MO on vacuolar hepatocyte degeneration exceeded that of the MSC on d14 and both MS and MSC on d42. Each treatment was equally effective in the decrease of the severity of solitary cell death and infiltration of lympho- and histiocytes in the liver on d28 as well as in the prevention of lymphocyte depletion in the spleen and bursa of Fabricius on d14. In conclusion, the applied treatments have been proven effective in the prevention of histopathological changes caused by DON and ZEN.

Comparing effects of natural betaine and betaine hydrochloride on gut physiology in broiler chickens.

Betaine is a well-known component of poultry diets with various effects on nutritional physiology. For example, increased water retention due to the osmolytic effect of betaine increases the volume of the cell, thereby accelerating the anabolic activity, integrity of cell membrane, and overall performance of the bird. Betaine is a multifunctional component (trimethyl derivative) acting as the most efficient methyl group donor and as an organic osmolyte, which can directly influence the gastrointestinal tract integrity, functionality, and health. So far, nothing is known about the effect of betaine on the intestinal barrier in chickens. In addition, little is known about comparing natural betaine with its synthetic form. Therefore, an animal study was conducted to ascertain the effects of betaine supplementation (natural and synthetic) on performance and intestinal physiological responses of broilers. One hundred and five 1-day-old broiler chicks were randomly assigned into 3 groups with 35 birds each: control, natural betaine (1 kg active natural (n)-betaine/ton of feed) and synthetic (syn)-betaine-HCL (1 kg active betaine /ton of feed). Histological assessment showed lower jejunal crypt depth and villi height/crypt depth ratio in syn-betaine-HCL group compared with natural n-betaine fed birds. Furthermore, it was found that syn-betaine-HCL negatively affects the integrity of the intestine by increasing the intestinal paracellular permeability in both jejunum and cecum as evidenced by a higher mannitol flux. Additionally, syn-betaine-HCl significantly upregulated the IFN-γ mRNA expression at certain time points, which could promote intestinal permeability, as it plays an important role in intestinal barrier dysfunction. Body weight (BW) and body weight gain (BWG) did not differ (P > 0.05) between the control birds and birds supplemented with syn-betaine-HCL. However, the BW and BWG were significantly (P < 0.05) improved by the dietary inclusion of n-betaine compared with other treatments. Altogether, the dietary inclusion of n-betaine had a positive effect on performance and did not negatively affect gut paracellular permeability. Furthermore, our results show that syn-betaine-HCl induces changes in the intestine, indicating an alteration of the intestinal histology and permeability. Thus, natural or synthetic betaine has different effects, which needs to be considered when using them as a feed supplement.

Microbiota Composition of Mucosa and Interactions between the Microbes of the Different Gut Segments Could Be a Factor to Modulate the Growth Rate of Broiler Chickens.

The study reported here aimed to determine whether correlations can be found between the intestinal segment-related microbiota composition and the different growing intensities of broiler chickens. The bacterial community structures of three intestinal segments (jejunum chymus-JC, jejunum mucosa-JM, caecum chymus-CC) from broiler chickens with low body weight (LBW) and high body weight (HBW) were investigated. Similar to the previous results in most cases, significant differences were found in the bacteriota diversity and composition between the different sampling places. However, fewer body weight (BW)-related differences were detected. In the JM of the HBW birds, the Bacteroidetes/Firmicutes ratio (B/F) was also higher. At the genus level significant differences were observed between the BW groups in the relative abundance of Enterococcus, mainly in the JC; Bacteroides and Ruminococcaceae UCG-010, mainly in the JM; and Ruminococcaceae UCG-013, Negativibacillus, and Alistipes in the CC. These genera and others (e.g., Parabacteroides and Fournierella in the JM; Butyricoccus, Ruminiclostridium-9, and Bilophila in the CC) showed a close correlation with BW. The co-occurrence interaction results in the JC revealed a correlation between the genera of Actinobacteria (mainly with Corynebacterium) and Firmicutes Bacilli classes with different patterns in the two BW groups. In the JM of LBW birds, two co-occurring communities were found that were not identifiable in HBW chickens and their members belonged to the families of Ruminococcaceae and Lachnospiraceae. In the frame of the co-occurrence evaluation between the jejunal content and mucosa, the two genera (Trichococcus and Oligella) in the JC were found to have a significant positive correlation with other genera of the JM only in LBW chickens.

The relationship between small heat shock proteins and redox homeostasis during acute heat stress in chickens.

As heat stress is a major emerging issue in poultry farming, investigations on the molecular mechanisms of the heat-triggered cellular response in chickens are of special importance. In the present study, 32-day-old Ross 308 broiler chickens were subjected to 37 °C environmental temperature combined with 50% relative humidity for 4 or 8 h respectively. Following sampling, redox parameters such as malondialdehyde (MDA), reduced glutathione (GSH), protein carbonyl levels as well as glutathione peroxidase activity were assessed in liver, spleen, and kidney homogenates. The concentrations of small heat shock proteins (sHSP-s) HSP27, αA- and αB-crystallins were also investigated. Among these organs, the liver was found the most susceptible to heat-provoked oxidative stress, indicated by enhanced lipid peroxidation and rapid activation of protective pathways, including the definite increase of glutathione peroxidase activity and the excessive utilization of αA- and αB-crystallin proteins. Heat-associated decline of protein carbonylation and GSH content was observed in the liver in correlation with the increased involvement of αA- and αB-crystallins in cellular defense, resulting supposedly in an overcompensation mechanism. These data highlight the hepatic sensitivity to acute heat shock, potential adaptation mechanisms, and the specific role of sHSP-s in the restoration of physiologic cell function.

Effects of Probiotics and Wheat Bran Supplementation of Broiler Diets on the Ammonia Emission from Excreta.

Ammonia emission is a concern for the poultry industry from both environmental and animal welfare points of view. The objective of this research was to determine whether probiotics or wheat bran supplementation of broiler diets can modify the N composition of the excreta and the dynamics of ammonia volatilisation emission from the manure. A total of 120-day-old Ross 308 broiler chickens were fed six different diets. The treatments included a corn and soybean meal-based control diet (C) and diets containing wheat bran (WB). Both diets were fed alone and with supplementation of a lactic acid (Lactobacillus farciminis, LAB) and a butyric acid (Clostridium butyricum, BAB) producing bacterial strain. Treatment BAB had a significant effect on the dry matter content of the excreta and both probiotics decreased the amount of excreted uric acid. Treatment WB resulted in a significantly lower NH+4-N concentration of excreta and a tendency toward reduced uric acid content. Treatment LAB reduced the urinary N ratio of excreta. Among dietary treatments, WB resulted in the highest urease producing cell numbers in the excreta, but this difference was not significant. Based on our results, similar to pigs, the soluble fibre fraction of poultry diets can also modify the urinary to faecal N ratio of the excreta.

Relative Effects of Dietary Administration of a Competitive Exclusion Culture and a Synbiotic Product, Age and Sampling Site on Intestinal Microbiota Maturation in Broiler Chickens.

In this research, the effects of early post-hatch inoculation of a competitive exclusion product (Br) and the continuous feeding of a synbiotic supplement (Sy) containing probiotic bacteria, yeast, and inulin on the production traits and composition of ileal chymus (IC), ileal mucosa (IM), and caecal chymus (CC) microbiota of broiler chickens were evaluated. The dietary treatments had no significant effects on the pattern of intestinal microbiota or production traits. The digestive tract bacteriota composition was affected mostly by the sampling place and age of birds. The dominant family of IC was Lactobacillaceae, without change with the age. The abundance of the two other major families, Enterococcaceae and Lachnospiraceae decreased with the age of birds. In the IM, Clostridiaceae was the main family in the first three weeks. Its ratio decreased later and Lactobacillaceae became the dominant family. In the CC, Ruminococcaceae and Lachnospiraceae were the main families with decreasing tendency in the age. In IC, Br treatment decreased the abundance of genus Lactobacillus, and both Br and Sy increased the ratio of Enterococcus at day 7. In all gut segments, a negative correlation was found between the IBD antibody titer levels and the ratio of genus Leuconostoc in the first three weeks, and a positive correlation was found in the case of Bifidobacterium, Rombutsia, and Turicibacter between day 21 and 40.

Effects of Wheat Bran and Clostridium butyricum Supplementation on Cecal Microbiota, Short-Chain Fatty Acid Concentration, pH and Histomorphometry in Broiler Chickens.

Feed additives that can improve intestinal health and maintain a diverse and resilient intestinal microbiota of poultry are of great importance. Thus, the current study investigated the effects of a single strain butyric acid-producing Clostridium (C. butyricum) with (symbiotic) or without wheat bran supplementation on cecal microbiota composition and gut health characteristics of broiler chickens. In total, 384 male Ross 308 day-old chickens were divided into four dietary treatment groups and fed ad libitum until day 37 of life. Cecal samples were taken for Illumina sequencing and pH and short-chain fatty acid analyses, as well as for histological analysis at the end of the experimental period. Neither of the supplemented diets improved chicken growth performance. Caecum was dominated by the members of Bacteroidetes phyla followed by Firmicutes in each dietary group. At the genus level, Bacteroides, Oscillospira, Akkermansia, Faecalibacterium, Ruminococcus and Streptococcus genera exceeded 1% relative abundance. Dietary treatment influenced the relative abundance of the Akkermansia genus, which had a lower relative abundance in the C. butyricum group than in the other groups and in the symbiotic group compared to the wheat bran supplemented group. Dietary treatment also altered cecal crypt depth and had a trend to modify the cecal fermentation profile. Additive effects of wheat bran and C. butyricum supplementation were not detected. Our results suggest that Akkermansia muciniphila colonization in chicken can be influenced by diet composition.

Effects of Acute Heat Stress on a Newly Established Chicken Hepatocyte-Nonparenchymal Cell Co-Culture Model.

Heat stress is one of the most important issues in broiler flocks impairing animal health and productivity. On a cellular level, excess heat exposure can trigger heat shock response acting for the restoration of cell homeostasis by several mechanisms, such as affecting heat shock protein synthesis, redox homeostasis and pro-inflammatory cytokine production. The major aim of this study was to establish a novel avian hepatocyte-nonparenchymal cell co-culture as a model for investigating the cellular effects of heat stress and its interaction with inflammation in chicken liver. Cell fractions were isolated by differential centrifugation from a freshly perfused chicken liver, and hepatocyte mono-cultures as well as hepatocyte-nonparenchymal cell co-cultures (with cell ratio 6:1, hepatocytes to nonparenchymal cells, mimicking a milder hepatic inflammation) were prepared. Isolated and cultured cells were characterized by flow cytometry and immunocytochemistry applying hepatocyte- and macrophage-specific antibodies. Confluent cell cultures were exposed to 43 °C temperature for 1 or 2 h, while controls were cultured at 38.5 °C. The metabolic activity, LDH enzyme activity, reactive oxygen species (H2O2) production, extracellular concentration of heat shock protein 70 (HSP70), and that of the pro-inflammatory cytokines interleukin (IL-)6 and IL-8 were assessed. Shorter heat stress applied for 1 h could strongly influence liver cell function by significantly increasing catabolic metabolism and extracellular H2O2 release, and by significantly decreasing HSP70, IL-6, and IL-8 production on both cell culture models. However, all these alterations were restored after 2 h heat exposure, indicating a fast recovery of liver cells. Hepatocyte mono-cultures and hepatocyte-nonparenchymal cell co-cultures responded to heat stress in a similar manner, but the higher metabolic rate of co-cultured cells may have contributed to a better capability of inflamed liver cells for accommodation to stress conditions. In conclusion, the established new primary cell culture models provide suitable tools for studying the hepatic inflammatory and stress response. The results of this study highlight the impact of short-term heat stress on the liver in chickens, underline the mediatory role of oxidative stress in acute stress response, and suggest a fast cellular adaptation potential in liver cells.

Cellular Effects of T-2 Toxin on Primary Hepatic Cell Culture Models of Chickens.

Trichothecene mycotoxins such as T-2 toxin cause severe problems for agriculture, as well as for veterinary medicine. As liver is one of the key organs in metabolism, the main aim of our study was to investigate the immunomodulatory and cytotoxic effects of T-2 toxin, using primary hepatocyte mono-culture and hepatocyte-nonparenchymal cell (predominantly Kupffer cell) co-culture models of chicken. Cultures were exposed to 10 (T10 group), 100 (T100 group) and 1000 (T1000 group) nmol/L T-2 toxin treatment for 8 or 24 h. Alterations of cellular metabolic activity, the production of reactive oxygen species (extracellular H2O2), heat shock protein 70 (HSP70), and the concentration of different inflammatory cytokines such as interleukin (IL-)6 and IL-8 were investigated. Metabolic activity was intensely decreased by T-2 toxin administration in all of the cell culture models, in every applied concentration and incubation time. Concentrations of HSP70 and IL-8 were significantly increased in hepatocyte mono-cultures exposed to higher T-2 toxin levels (both in T100 and T1000 groups for HSP70 and in T1000 group for IL-8, respectively) compared to controls after 24 h incubation. Similarly, IL-6 levels were also significantly elevated in the T100 and T1000 groups in both of mono- and co-cultures, but only after 8 h of incubation time. In spite of the general harmful effects of T-2 toxin treatment, no significant differences were observed on reactive oxygen species production. Furthermore, the two cell culture models showed different levels of H2O2, HSP70, and IL-8 concentrations independently of T-2 toxin supplementation. In conclusion, the established primary cell cultures derived from chicken proved to be proper models to study the specific molecular effects caused by T-2 toxin. Metabolic activity and immune status of the different examined cell cultures were intensively affected; however, no changes were found in H2O2 levels.

Soluble nondigestible carbohydrates improve intestinal function and increase caecal coliform load in broiler chickens.

Diets rich in various soluble nondigestible carbohydrates (sNDCs) were evaluated on different intestinal characteristics (histological, physico-chemical and microbiological) of chickens and compared with a maize-based diet as a control. A total of 160 Ross 308 male chickens were kept in deep litter pens (n = 40) and fed their appropriate diets from Day 1 to Day 35 of life. Four isocaloric and isonitrogenous diets, differing in their sNDC content, were composed; control (containing maize as the only cereal), maize-wheat-based (M + W) and maize-based supplemented with either 20 g/kg inulin (M + I) or 30 g/kg lactose (M + L). All of the diets tested decreased ileal crypt depth, ileal muscle layer thickness and increased caecal coliform counts relative to the control group. Villus-crypt ratio increased only in the M + L group. Ileal digesta of chickens fed the M + W diet had the highest ileal viscosity and the highest caecal butyrate, valerate and total short-chain fatty acid concentrations while the lowest pH was observed in caecal contents of chickens fed the M + I diet. The diet had no effect on ileal or caecal goblet cell and intraepithelial lymphocyte numbers. Lactobacillus counts in the caecal content remained unchanged. According to the present study, various sNDC sources may have beneficial gut health effects, however, some of the intestinal variables are dependent on the type of sNDCs.