Dietary Peppermint Extract Inhibits Chronic Heat Stress-Induced Activation of Innate Immunity and Inflammatory Response in the Spleen of Broiler Chickens.

The aim of this study was to investigate the effect of dietary peppermint extract (PE) on innate immunity and inflammatory responses in the spleen of broiler chickens under chronic heat stress. In order to further study the mechanism of the activation of innate immunity and inflammation induced by chronic heat stress and the regulatory effect of peppermint extract, we examined the spleen's histological change, the mRNA expression of major pattern recognition receptors (PRRs) (TLR2, TLR4, NOD1, MDA5 and DAI) and transcription factors (NF-κB, AP-1 and IRF3) and downstream inflammatory cytokines (IFN-α, IFN-ß, IL-1ß, IL-6 and TNF-α) of innate immune signaling pathways associated with heat stress in the spleen of broiler chickens. The results indicated that chronic heat stress damaged the spleen tissue. In addition, chronic heat stress induced the activation of innate immunity and inflammatory responses by increasing the mRNA expression of TLR2, TLR4 and DAI, mRNA expression of transcriptional factors (NF-κB, AP-1 and IRF3) and the concentration of downstream inflammatory cytokines in the spleen of broiler chickens. Dietary peppermint extract alleviated the damage of spleen tissue caused by chronic heat stress. In addition, peppermint extract reduced the mRNA expression of DAI, mRNA expression of transcriptional factors NF-κB, AP-1 and IRF3, and the concentration of inflammatory cytokines in the spleen of broiler chickens under chronic heat stress. In conclusion, dietary peppermint extract could have a beneficial effect on regulating inflammatory response and innate immunity via inhibiting the activation of NF-κB, AP-1 and IRF3 signaling pathways mediated by DAI in the spleen of broiler chickens induced by chronic heat stress.

Research Note: The effect of photoperiod on the NLRP3 inflammasome and gut microbiota in broiler chickens.

The present study aimed to investigate the effect of photoperiod on the intestinal inflammation and gut microbiota. A total of 96 broiler chickens were divided into 2 groups and fed separately under 2 different photoperiods (12L:12D group and 23L:1D group) for 21 d. The results showed that the photoperiod of 23L:1D damaged duodenal tissue structure (intestinal villus erosion, mucosal epithelial cell detachment, and inflammatory cell infiltration), significantly increased the concentration of inflammatory cytokines (IL-1ß, IL-18, IL-6, and TNF-α) and significantly increased the mRNA expression levels and protein expression levels of NOD-, LRR-, pyrin domain-containing protein 3 (NLRP3) and caspase1 (P <0.05) compared with 12L:12D, which indicating that extended photoperiod induced intestinal injury and activated NLRP3 inflammasome. 16S rRNA sequencing analysis revealed that Bacteroides was significantly decreased, Ruminococcus_torques_group, norank_f_Desulfovibrionaceae, GCA-900066575, Defluviitaleaceae_UCG-011, Lachnospiraceae_FCS020_group, norank_f_UCG-010 and norank_f_norank_o_Clostridia_vadinBB60_group and were significantly increased in the 23L:1D group, compared with the 12L:12D group (P < 0.05). The correlation analysis between differential microbial communities and intestinal inflammation showed that the relative abundance of Bacteroides was negatively correlated with the mRNA expression level of NLRP3 (P < 0.05) and the relative abundance of Ruminococcus_torques_group was positively correlated with the mRNA expression level of NLRP3 (P < 0.05). linear discriminant analysis (LDA) effect size (LEfSe) results (LDA > 4) showed that the relative abundance of Bacteroides was dramatically higher (P < 0.05) in the 12L:12D group, whereas the relative abundance of Ruminococcus_torques_group was noticeably higher (P < 0.05) in the 23L:1D group. By the comprehensive analysis of the gut microbiota, the interaction of gut microbiota (Bacteroides and Ruminococcus_torques_group) and NLRP3 inflammasome may contribute to the intestinal injury under the condition of extended photoperiod.

Dietary tryptophan alleviates intestinal inflammation caused by long photoperiod via gut microbiota derived tryptophan metabolites-NLRP3 pathway in broiler chickens.

Light pollution is a potential risk factor for intestinal health. Tryptophan plays an important role in the inhibition of intestinal inflammation. However, the mechanism of tryptophan in alleviating intestinal inflammation caused by long photoperiod is still unclear. This study investigated the anti-inflammatory effect of dietary tryptophan on intestinal inflammatory damage induced by long photoperiod and its potential mechanism in broiler chickens. We found that dietary tryptophan mitigated long photoperiod-induced intestinal tissue inflammatory damage and inhibited the activation of Nucleotide-Binding Oligomerization Domain, Leucine-Rich Repeat and Pyrin Domain-Containing 3 inflammasome. Moreover, dietary tryptophan significantly increased the relative abundance of Faecalibacterium, Enterococcus, and Lachnospiraceae_NC2004_group were significantly decreased the relative abundance of Ruminococcus_torques_group and norank_f_UCG-010 under the condition of long photoperiod (P < 0.05). The results of tryptophan targeted metabolomics show that tryptophan significantly increased indole-3-acetic acid (IAA) and indole-3 lactic acid (ILA), and significantly decreased xanthurenic acid (XA) under long photoperiod (P < 0.05). In conclusion, the results indicated that dietary tryptophan alleviates intestinal inflammatory damage caused by long photoperiod via the inhibition of Nucleotide-Binding Oligomerization Domain, Leucine-Rich Repeat and Pyrin Domain-Containing 3 inflammasome activation, which was mediated by tryptophan metabolites. Therefore, tryptophan supplementation could be a promising way to protect the intestine health under the condition of long photoperiod.

The amino acid pattern and dynamics of body protein, body fat deposition in male and female broilers under different temperatures.

The present study was conducted 1) to investigate the effects of gender and temperature on growth performance in broiler chickens and 2) to establish body protein and fat deposition curves and amino acid patterns for broilers of both genders at different ambient temperatures. A total of 432 1-day-old (d) Arbor Acres chickens with a male/female ratio of 1:1 were randomly divided into the following 4 treatment groups: the male thermoneutral group, the female thermoneutral group, the male heat stress group, and the female heat stress group. The chickens in the thermoneutral groups were kept at a comfortable temperature from 1 to 42 d, while chickens in the heat stress groups were kept at a comfortable temperature from 1 to 28 d and at a high ambient temperature from d 29 to 42. The body composition retention data were obtained by comparative slaughter method, and the models were constructed by the Gompertz model. The results revealed significant variation in body protein content (BPC) and body fat deposition efficiency (BFE) between both genders and the 2 temperatures. Moreover, a noteworthy interaction between gender and temperature was observed in terms of the BPC and protein deposition efficiency (BPE). The following equations for body protein and body fat deposition in the thermoneutral groups were obtained: Body protein weight of male broilers: [Formula: see text] ; Body protein weight of female broilers: [Formula: see text] ; Body fat weight of male broilers: [Formula: see text] ; Body fat weight of female broilers: [Formula: see text] . Where t means age (d). The following equations for body protein and body fat deposition in the heat stress groups were obtained: Body protein weight of male broilers: [Formula: see text] ; Body protein weight of female broilers: [Formula: see text] ; Body fat weight of male broilers: [Formula: see text] ; Body fat weight of female broilers: [Formula: see text] . Where t means age (d). In addition, no significant difference in amino acid content was found between different genders and temperatures. The amino acid pattern could be divided into 2 stages: 0 to 14 d and 15 to 42 d. Our equations and patterns enable a deeper understanding of the nutritional requirements in broiler chickens under various temperature conditions. This enables researchers to develop more accurate feeding programs to fulfill the growth and health requirements of broiler chickens.

Effects of Heat Stress on Breast Muscle Metabolomics and Lipid Metabolism Related Genes in Growing Broilers.

With global warming and worsening climatic conditions, heat stress (HS) has become a significant challenge affecting the development of poultry production. In this study, we aimed to determine the effects of HS on breast muscle metabolomics and lipid metabolism-related genes in growing broilers. One hundred twenty 29-day-old Arbor Acres broilers were randomly divided into normal temperature (NT; 21 ± 1 °C) and heat stress (HS; 31 ± 1 °C) groups, with six replicates (ten birds in each replicate) in each group, raised for 14 days in two environment chambers at 60 ± 7% relative humidity. Compared with the broilers in the NT group, the average daily food intake, average daily gain and breast muscle yield in the HS group were significantly lower (p < 0.05). The feed conversion ratio was significantly higher in the HS group (p < 0.05). The concentrations of serum corticosterone, free fatty acids and cholesterol and the percentage of abdominal fat of broilers in the HS group were significantly higher (p < 0.05) than the values of the broilers in the NT group. Untargeted breast muscle metabolome analysis revealed 14 upregulated differential metabolites, including glycerophosphocholine, and 27 downregulated differential metabolites, including taurine, in the HS group compared to the NT group; the HS group also displayed significant effects on six metabolic pathways compared to the NT group (p < 0.05). The mRNA expression levels of peroxisome proliferator-activated receptor gamma coactivator-1-alpha, peroxisome proliferator-activated receptor alpha (PPARα) and ATP-binding cassette transporter A1 in the liver and breast muscles were significantly decreased in the HS group compared with the NT group (p < 0.05). The collective findings reveal that HS can cause disorders in breast muscle lipid metabolism in broilers. The PPARα gene might be the key gene in the mechanism of the lipid metabolism that is induced by HS in breast muscle of broilers. These findings provide novel insights into the effects of HS on chicken growth.

Effects of Atmospheric Ammonia on Skeletal Muscle Growth in Broilers.

Ammonia, one of the most polluted gases in poultry houses, has always been an urgent problem to solve. Exposure to ammonia can threaten the respiratory tract, induce inflammation, and decrease growth performance. To date, there are few studies investigating the effects of ammonia on skeletal muscle growth. In this experiment, a total of 144 broilers were randomly divided into two groups, and 0 ppm and 35 ppm atmospheric ammonia were administered in the chambers. The trial lasted for 21 days. The breast muscle, thigh muscle, dressed weight, and serum biochemical indexes were measured. The skeletal muscle fibre morphology was observed using light microscopy, and the expressions of genes associated with skeletal muscle development and myosin heavy chain genes were assessed. After 7 days of ammonia exposure, the broilers' weight in the ammonia group decreased. On the 21st day of the experiment, in the ammonia group, the breast muscle weight, thigh muscle weight, and dressed weight decreased, the blood urea nitrogen content increased, skeletal muscle fibre diameter shortened, the expression of myostatin increased, and the expression of myosin heavy chain-FWM and myosin heavy chain-FRM decreased significantly. This article suggests that 35 ppm atmospheric ammonia seriously affects the skeletal muscle gain rate of broilers, and the myostatin pathway could be a potential regulation of the growth rate of muscle fibre under ammonia exposure.

Interactive effects of high temperature and crude protein levels on growth performance, nitrogen excretion, and fecal characteristics of broilers.

In the present study, we aimed to explore the interactive effects of high temperature (HT) and dietary crude protein (CP) levels on nitrogen (N) excretion, fecal characteristics, and growth performance of broilers. A total of 288 broilers (Arbor Acres) were divided into six groups with eight replicates (six broilers per replicate). Two temperatures (ambient temperature: AT, 23 °C; HT: 28 ~ 32 ~ 28 °C) and three diets (CP: 14.90%, 18.18%, or 21.19%, with equal amounts of essential amino acids) were examined in a 2 × 3 factorial design. The experiment arrangement was from 4 to 6 weeks of age. The results showed that HT led to a significant decrease in the N excretion (P < 0.0001), average daily feed intake (P < 0.0001), and weight gain of broilers (P < 0.0001), while it markedly increased the fecal pH (P = 0.015), fecal moisture (P = 0.0014), uric acid (UA) contents (P = 0.0018), and feed/gain ratio (P < 0.0001). A low CP diet significantly decreased the N excretion (P < 0.001), fecal pH (P = 0.016), fecal moisture (P < 0.0001), and UA contents (P < 0.0001), while it markedly increased the feed/gain ratio (P < 0.001). In conclusion, HT had a negative impact on the fecal characteristics and growth performance of broilers but showed positive effects on N excretion. Moreover, decreased CP levels had a positive effect on the N excretion and fecal characteristics in broilers.

Gut microbiota dysbiosis exaggerates ammonia-induced tracheal injury Via TLR4 signaling pathway.

Ammonia is a toxic air pollutant that causes severe respiratory tract injury in animals and humans. Gut microbiota dysbiosis has been found to be involved in the development of respiratory tract injury induced by air pollutants, however, the specific mechanism requires investigation. Here, we found that, inhaled ammonia induced tracheal injury by reducing expression of claudin-1, increasing expression of muc5ac, TLR4, MyD88, NF-κB and cytokines (TNF-α, IL-1ß, IL-6 and IL-10), and also altering tracheal microbiota composition. Spearman correlation analysis indicated that gut microbiota dysbiosis positively correlated with TLR4 level in the trachea. Antibiotic depletion intestinal microbiota treatment reduced the severity of ammonia-induced tracheal injury via TLR4 signaling pathway. Microbiota transplantation induced the tracheal injury via TLR4 signaling pathway even without the ammonia exposure. These results indicate that gut microbiota dysbiosis exaggerates ammonia-induced tracheal injury via TLR4 signaling pathway. In addition, the [Ruminococcus]_torques_group, Faecalibacterium, unclassified_f_Lachnospiraceae may be the key gut microbiota contributing to the alterations of tracheal microbiota composition.

Effects of Increasing Stocking Density on the Performance and Ileal Microbiota of Broilers.

This study was conducted to investigate the effects of increasing stocking density under suitable environmental conditions on the performance and ileal microbiota of broilers. A total of 108 Arbor Acres male broilers (28 days old) were allocated to a normal stocking density (NSD, normal stocking density; 31 kg/m2) and a maximum allowed stocking density group (MSD, maximum stocking density; 39 kg/m2). All birds were reared at a constant temperature of 21°C. At 42 days of age, bacterial DNA was extracted from ileal content, and the V3-4 hypervariable region of 16S rRNA was amplified. Increasing stocking density had no significant effect on average daily gain, average daily feed intake, and feed conversion ratio (P>0.05). The alpha and beta diversities of the ileal microbiomes did not differ significantly between the NSD and MSD groups; however, increasing stocking density altered the composition of ileal microbiota. The relative abundance of Lactobacillales, including Lactobacillus, Enterococcus, and Streptococcus, significantly decreased in MSD broilers, compared with NSD broilers. The present results suggest that even under suitable environmental conditions, an increase in stocking density to a level of 39 kg/m2 may disturb the composition of ileal microbiota in broilers. Further studies are needed to determine the reasons and the potential consequences for animal health and physiology.

Effects of Dietary Chromium Picolinate on Gut Microbiota, Gastrointestinal Peptides, Glucose Homeostasis, and Performance of Heat-Stressed Broilers.

The current research was devoted to evaluating the effects on gut microbiota, gastrointestinal peptides, and glucose homeostasis of chromium picolinate applied to heat-stressed broilers. In a 14 d experiment, 220 28-day-old AA broilers were randomly assigned into one thermal-neutral and three high-temperature groups dietary-supplemented with 0, 0.4, or 0.8 mg/kg of chromium as chromium picolinate. The temperature for the thermal-neutral group was set at 21 °C, while that for the other three groups (high temperature) was set at 31 °C. The results showed that the average daily gain and average daily feed intake of the 0.4 mg/kg chromium-supplemented group significantly increased compared with the high-temperature groups (p < 0.05). The content of cholecystokinin in the 0.4 mg/kg group significantly decreased, and the gastric inhibitory polypeptide level was significantly elevated in jejunum (p < 0.05). The cecal microbiota of heat-stressed broilers was substantially different from that of the thermal-neutral group. After diet-supplemented chromium, compared to the high-temperature groups, the 0.4 mg/kg chromium supplemented group was characterized by a reduction of Actinobacteriota and Proteobacteria at the phylum level. The Bacilli were elevated, while proportions of Coriobacteria and Gammaproteobacteria were reduced significantly at the class level. The proportions of Lactobacillaceae, Christensenellaceae, and Erysipelotrichaceae were elevated significantly, while that of Clostridiaceae was reduced significantly at the family level. The proportion of Turicibacter was elevated significantly and the proportions of Olsenella and Ruminococcus were reduced significantly at the genus level (p < 0.05). Compared to the high-temperature groups, in the 0.4 mg/kg chromium-supplemented group, the insulin concentration and insulin resistance index were reduced (p < 0.05), and sodium-glucose transporter 1 expression was up-regulated in jejunum (p < 0.05). Performance, microbiota, gastrointestinal peptides, or serum parameters of the 0.8 mg/kg group were almost unaffected by chromium compared with the high-temperature groups. In conclusion, diet supplemented with 0.4 mg/kg Cr improved performance, insulin resistance and sodium-glucose transporter 1 expression and altered gut microflora structure and secretion of gastrointestinal peptides, thus showing that supplementation with chromium is beneficial to maintain glucose homeostasis and alleviate heat stress.

Effects of Different Ammonia Concentrations on Pulmonary Microbial Flora, Lung Tissue Mucosal Morphology, Inflammatory Cytokines, and Neurotransmitters of Broilers.

Atmospheric ammonia is one of the main environmental stressors affecting the performance of broilers. Previous studies demonstrated that high levels of ammonia altered pulmonary microbiota and induced inflammation. Research into the lung-brain axis has been increasing in recent years. However, the molecular mechanisms in pulmonary microbiota altered by ambient ammonia exposure on broilers and the relationship between microflora, inflammation, and neurotransmitters are still unknown. In this study, a total of 264 Arbor Acres commercial meal broilers (21 days old) were divided into 4 treatment groups (0, 15, 25, and 35 ppm group) with 6 replicates of 11 chickens for 21 days. At 7 and 21 D during the trial period, the lung tissue microflora was evaluated by 16S rDNA sequencing, and the content of cytokines (IL-1ß, IL-6, and IL-10) and norepinephrine (NE), 5-hydroxytryptamine (5-HT) in lung tissue were measured. Correlation analysis was established among lung tissue microflora diversity, inflammatory cytokines, and neurotransmitters. Results showed that the broilers were not influenced after exposure to 15 ppm ammonia, while underexposure of 25 and 35 ppm ammonia resulted in significant effects on pulmonary microflora, inflammatory cytokines, and neurotransmitters. After exposure to ammonia for 7 and 21 days, both increased the proportion of Proteobacteria phylum and the contents of IL-1ß and decreased the content of 5-HT. After exposure to ammonia for 7 days, the increase in Proteobacteria in lung tissue was accompanied by a decrease in 5-HT and an increase in IL-1ß. In conclusion, the microflora disturbance caused by the increase in Proteobacteria in lung tissue may be the main cause of the changes in inflammatory cytokines (IL-1ß) and neurotransmitters (5-HT), and the damage caused by ammonia to broiler lungs may be mediated by the lung-brain axis.

Ammonia exposure induced intestinal inflammation injury mediated by intestinal microbiota in broiler chickens via TLR4/TNF-α signaling pathway.

Ammonia is a known environmental pollutant that causes injury to the intestine. Growing evidence suggests that intestinal microbiota dysbiosis involves in the development of intestinal injury under environmental pollution. However, the specific mechanism remains unexplored. To do this, broiler chicken ileal exposed to ammonia was selected as the research object. Further, antibiotic depletion of intestinal microbiota and flora transplantation were used to clarify the role of intestinal microbiota in the intestinal injury. Histopathological examination indicated inhaled ammonia caused intestinal injury. Then we observed a decrease in intestinal muc-2, claudin-1, IL-6, IL-10 in ammonia inhalation, as opposed to the control group, associated with a significant increase in TLR4, MyD88, NF-κB, TNF-α, IL-1ß, caspase3. Moreover, there was a significant increase of Streptococcus, Escherichia-Shigella, Faecalibacterium, [Ruminococcus]_torques_group, Ruminococcaceae_UCG-014, unclassified_f_Lachnospiraceae, Rothia, unclassified_f_Ruminococcaceae in the inhaled ammonia exposure. Correlation analysis suggested that the altered genera were positively correlated with the expression of TLR4 and TNF-α. Moreover, transferring intestinal microbiota from ammonia exposure broiler into healthy broiler caused intestinal injury and increased TLR4 and TNF-α concentrations in recipient broiler. Furthermore, antibiotic depletion of intestinal microbiota attenuated ammonia-caused intestinal injury and reduced TLR4 and TNF-α productions. In summary, TLR4/TNF-α signaling pathway was an important regulated mechanism involved in the intestinal injury mediated by intestinal microbiota dysbiosis under inhaled ammonia.

Effects of Ammonia on Gut Microbiota and Growth Performance of Broiler Chickens.

In order to investigate the influence of ammonia on broiler intestinal microflora and growth performance of broiler chickens, 288 21-day-old male Arbor Acres broilers with a similar weight were randomly divided into four groups with different NH3 levels: 0 ppm, 15 ppm, 25 ppm, and 35 ppm. The growth performance of each group was recorded and analyzed. Additionally, 16s rRNA sequencing was performed on the cecal contents of the 0 ppm group and the 35 ppm group broilers. The results showed the following: a decrease in growth performance in broilers was observed after 35 ppm ammonia exposure for 7 days and 25 ppm ammonia exposure for 14 days. At phylum level, the relative abundance of Proteobacteria phylum was increased after 35 ppm ammonia exposure. At genus level, ammonia increased the relative abundance of Escherichia-Shigella and decreased the relative abundance of Butyricicoccus, Parasutterella, Lachnospiraceae_UCG-010, Ruminococcaceae_UCG-013 and Ruminococcaceae_UCG-004. Negative correlation between Escherichia-Shigella and growth performance, and positive correlation between bacteria genera (including Butyricicoccus, Parasutterella, Lachnospiraceae_UCG-010, Ruminococcaceae_UCG-013 and Ruminococcaceae_UCG-004) and growth performance was observed. In conclusion, ammonia exposure caused changes in the structure of cecal microflora, and several species were either positively or negatively correlated with growth performance. These findings will help enhance our understanding of the possible mechanism by which ammonia affect the growth of broilers.

Myostatin and Related Factors Are Involved in Skeletal Muscle Protein Breakdown in Growing Broilers Exposed to Constant Heat Stress.

Heat stress has an adverse effect on the development of poultry farming, which has always aroused great concern. This study was carried out to investigate the protein breakdown mechanism responsible for the suppressive effect of constant heat stress on muscle growth in growing broilers. A total of 96, 29-day-old, Arbor Acres male broilers were randomly divided into two groups, a thermoneutral control (21 ± 1 °C, TC) and a heat stress (31 ± 1 °C, HS) group, with six replicates in each group and eight birds in each replicate. The trial period lasted for 14 d, and the trial was performed at 60 ± 7% relative humidity, a wind speed of <0.5 m/s and an ammonia level of <5 ppm. The results showed that the average daily feed intake and average daily gain in the HS group were distinctly lower than those in the TC group (p < 0.05), whereas the HS group showed a significantly increased feed conversion ratio, nitrogen excretion per weight gain and nitrogen excretion per feed intake compared to the TC group (p < 0.05). In addition, the HS group showed a significantly reduced breast muscle yield and nitrogen utilization in the broilers (p < 0.05). The HS group showed an increase in the serum corticosterone level (p < 0.05) and a decrease in the thyroxine levels in the broiler chickens (p < 0.05) compared to the TC group, whereas the HS group showed no significant changes in the serum 3,5,3'-triiodothyronine levels compared to the TC group (p > 0.05). Moreover, the HS group showed increased mRNA expression levels of myostatin, Smad3, forkhead box O 4, muscle atrophy F-box and muscle ring-finger 1, but reduced mRNA expression levels of the mammalian target of rapamycin, the protein kinase B and the myogenic determination factor 1 (p < 0.05). In conclusion, the poor growth performance of birds under constant heat stress may be due to an increased protein breakdown via an mRNA expression of myostatin and related factors.

Effects of Heat Stress on Gut-Microbial Metabolites, Gastrointestinal Peptides, Glycolipid Metabolism, and Performance of Broilers.

This paper investigated the effects of heat stress on gut-microbial metabolites, gastrointestinal peptides, glycolipid metabolism, and performance of broilers. Thus, 132 male Arbor Acres broilers, 28-days-old, were randomly distributed to undergo two treatments: thermoneutral control (TC, 21 °C) and high temperature (HT, 31 °C). The results showed that the average daily gain (ADG), average daily feed intake (ADFI), and gastric inhibitory polypeptide (GIP) concentration in the jejunum significantly decreased the core temperature, feed conversion ratio (FCR), and ghrelin of the hypothalamus, and cholecystokinin (CCK) in jejunum, and serum significantly increased in the HT group (p < 0.05). Exploration of the structure of cecal microbes was accomplished by sequencing 16S rRNA genes. The sequencing results showed that the proportion of Christensenellaceae and Lachnospiraceae decreased significantly whereas the proportion of Peptococcaceae increased at the family level (p < 0.05). Ruminococcus and Clostridium abundances significantly increased at the genus level. Furthermore, the content of acetate in the HT group significantly increased. Biochemical parameters showed that the blood glucose concentration of the HT group significantly decreased, and the TG (serum triglycerides), TC (total cholesterol), insulin concentration, and the insulin resistance index significantly increased. Nonesterified fatty acid (NEFA) in the HT group decreased significantly. In conclusion, the results of this paper suggest that the poor production performance of broilers under heat stress may be related to short-chain fatty acids (SCFAs) fermented by intestinal microbiota involved in regulating metabolic disorders.

The alterations of tracheal microbiota and inflammation caused by different levels of ammonia exposure in broiler chickens.

Ammonia (NH3) is a known harmful gas and exists in haze, forming secondary organic aerosols. Exposure to ambient ammonia correlates with the respiratory tract infection, and microbiota in the upper respiratory tract is an emerging crucial player in the homeostatic regulation of respiratory tract infection, and microbiota perturbation is usually accompanied by the inflammatory reactions; however, the effects of different levels of ammonia exposure on tracheal microbiota and inflammation are unclear. A total of 288 22-day-old male Arbor Acres broilers were chosen and divided into 4 groups with 6 replicates of 12 chickens, and respectively exposed to ammonia at 0, 15, 25, and 35 ppm for 21-d trial period. Cytokines (interleukin (IL)-1ß, IL-6, and IL-10) in the trachea were measured at the 21 d of exposure to NH3. Tracheal microbiota at the 21 d was analyzed by the 16S rRNA gene analysis. The results showed that an increase in ammonia levels, even in 15 ppm, significantly decreased the alpha diversity and changed the bacterial community structure. Six genera (Faecalibacterium, Ruminococcus]_torques_group, unclassified_f__Lachnospiraceae, Ruminococcaceae_UCG-014, Streptococcus, Blautia) significantly increased, whereas Lactobacillus significantly decreased under different levels of ammonia exposure. We also observed positive associations of Faecalibacterium, Blautia, g__Ruminococcaceae_UCG-014, unclassified_f__Lachnospiraceae and Ruminococcus]_torques_group abundances with tracheal IL-1ß concentration. Moreover, an increase in ammonia levels, even in 15 ppm, caused respiratory tract inflammatory injury. The results indicated that 15 ppm ammonia exposure changed the composition of tracheal microbiota that caused the tracheal injury possibly through increasing the IL-1ß, which might make the broiler more sensitive to the changes of environment and pathogenic micro-organisms in the poultry house, and may be also a critical value that needs high alertness. Herein, the present experiment also suggested that the standard limit of ammonia concentration in adult poultry house is 15 ppm. This research provides an insight into the relationship between the upper respiratory tract microbiota and inflammation under ammonia exposure.

Changes in the cecal microbiota of laying hens during heat stress is mainly associated with reduced feed intake.

The aim of the present study was to examine the effect of high temperature on the cecal microbiota of laying hens and analyze the correlation between variation in feed intake, body temperature, and the relative abundance of specific bacterial genera. Two hundred and sixteen 28-wk-old Hy-Line laying hens were randomly divided into 3 groups with 6 replicates and raised in 3 controlled climate chambers. Birds were allocated to either a thermoneutral group (TC, 21 ± 1°C, ad libitum), high cyclic temperature group (HT, 29-35°C, ad libitum), or a pair-feeding group (PF, 21 ± 1°C, pair-feeding on the daily feed intake of HT hens) for 4 wk. There was no significant difference in diversity estimators of laying hens between the HT and TC or PF groups (P > 0.05). Principal component analysis (PCoA) based on unweighted UniFrac distance showed that the HT group did not form a distinct cluster that was clearly separated from the PF group, but was separated from the TC group. Similarity analysis (ANOSIM) confirmed that the TC group was significantly different from the HT (R = 0.259, P = 0.020) and the PF (R = 0.348, P = 0.011) groups, but the HT group was similar with the PF group (R = 0.107, P = 0.153). In addition, there was a significant correlation between feed intake and principal coordinate 1 (PC1) in PCoA (R = -0.494, P = 0.037), but no correlation between core temperature and PC1 (R = 0.025, P = 0.923). Linear discriminant analysis identified 15 genera that discriminated between the HT and TC groups (score ≥2). Among these, the feed intake was significantly related with the relative abundance of 10 genera, however, the core temperature was only significantly correlated with one specific genus. In conclusion, a high cyclic temperature of 29-35°C had significant effects on microbial composition and structure of laying hens, which was mainly related with decreased feed intake.

iTRAQ-based quantitative proteomics analysis of the spleen reveals innate immunity and cell death pathways associated with heat stress in broilers (Gallus gallus).

Heat stress induces immune dysfunction and cell death, but the mechanisms by which this occurs are not fully understood. Therefore, the isobaric tags for relative and absolute quantification (iTRAQ) was used to identify differentially abundant proteins (DAPs) in the spleen between heat-stressed group and control group, and real time qPCR (RT-qPCR), and Parallel Reaction Monitoring (PRM) were performed to validate the differentially abundant proteins of interest. The results showed that nine down regulated DAPs related to innate immunity were enriched in the Toll-like receptor signaling pathway (IRF3 and CD40), NOD-like receptor signaling pathway (TNFAIP3, IL-18, CathL2, IRF3, IAP3 and CYBA), RIG-I-like receptor signaling pathway (TRIM25 and IRF3), and Cytosolic DNA-sensing pathway (IL-18, POLR3F and IRF3). Six down or up regulated DAPs related to cell death were enriched in apoptosis (CTSD, PARP3 and IAP3), ferroptosis (FTH) and necroptosis (FTH, CHMP1B, TNFAIP3, PARP3 and IAP3). In addition, compared with control group, heat stress significantly increased serums IL-1ß, IL-6, TNF-α, and IFN-α, as well as the splenocyte apoptosis rate, whereas significantly decreased serum IFN-ß. Taken together, these findings indicate that heat stress inhibits innate immunity and induces cell death through different pathways. SIGNIFICANCE: Our study identified potential signaling pathways and differentially abundant proteins related to the innate immunity and cell death of broilers under high temperature. These findings will facilitate a better understanding of the mechanisms of broiler response to heat stress and provide possible targets for alleviating heat stress in broiler production.

Probing the molecular regulation of lipopolysaccharide stress in piglet liver by comparative proteomics analysis.

Lipopolysaccharide (LPS) can induce inflammatory responses in piglets, causing immunological stress and tissue damage. However, chronic LPS infection may lead to LPS-induced immunological stress resistance. The molecular mechanisms underlying LPS stress have not been fully elucidated. Here, we conducted a global comparative proteomics analysis to investigate the molecular regulation of LPS stress using an immunological stress model of weaned piglets. A shotgun-based SWATH-MS workflow was used for global proteomes of the piglet livers after 15-day LPS treatment. Out of 3700 quantified proteins, 93 proteins showed differential changes under LPS stress. Bioinformatics analysis indicated that the differentially expressed proteins were mainly involved in inflammatory response, oxidation-redox processes and defense reactions, and were enriched in a phagosome pathway. Several key proteins associated with oxidative stress (SOD2), inflammation response (STEAP4 and S100 family) and the phagosome pathway were verified by activity and targeted-MS analyses. The observed responses appear to mitigate hepatic damage due to excessive oxidative stress, inflammation, and repression of the phagosome pathway. Our results reveal that an increased STEAP4 expression in piglets appears involved in cellular regulation by LPS stress and subsequent immunological stress resistance. This study sheds new light on the mechanism of prevention and relieving injury by LPS-induced immune responses.

Effects of a diet containing genetically modified rice expressing the Cry1Ab/1Ac protein (Bacillus thuringiensis toxin) on broiler chickens.

The aim of this study was to evaluate the effect of feeding Bacillus thuringiensis (Bt) rice expressing the Cry1Ab/1Ac protein on broiler chicken. The genetically modified (GM) Bt rice was compared with the corresponding non-GM rice regarding performance of feeding groups, their health status, relative organ weights, biochemical serum parameters and occurrence of Cry1Ab/1Ac gene fragments. One hundred and eighty day-old Arbor Acres female broilers with the same health condition were randomly allocated to the two treatments (6 replicate cages with 15 broilers in each cage per treatment). They received diets containing GM rice (GM group) or its parental non-GM rice (non-GM group) at 52-57% of the air-dried diet for 42 days. The results show that the transgenic rice had a similar nutrient composition as the non-GM rice and had no adverse effects on chicken growth, biochemical serum parameters and necropsy during the 42-day feeding period. In birds fed the GM rice, no transgenic gene fragments were detected in the samples of blood, liver, kidneys, spleen, jejunum, ileum, duodenum and muscle tissue. In conclusion, the results suggest that Bt rice expressing Cry1Ab/1Ac protein has no adverse effects on broiler chicken. Therefore, it can be considered as safe and used as feed source for broiler chicken.