Effects of Enteromorpha prolifera polysaccharides on growth performance,intestinal barrier function and cecal microbiota in yellow-feathered broilers under heat stress

Background Global warming leading to heat stress(HS)is becoming a major challenge for broiler production.This study aimed to explore the protective effects of seaweed(Enteromorpha prolifera)polysaccharides(EPS)on the intestinal barrier function,microbial ecology,and performance of broilers under HS.A total of 144 yellow-feathered broilers(male,56 days old)with 682.59±7.38 g were randomly assigned to 3 groups:1)TN(thermal neutral zone,23.6±1.8℃),2)HS(heat stress,33.2±1.5℃ for 10 h/d),and 3)HSE(HS+0.1%EPS).Each group contained 6 replicates with 8 broilers per replicate.The study was conducted for 4 weeks;feed intake and body weights were measured at the end of weeks 2 and 4.At the end of the feeding trial,small intestine samples were collected for histomorphology,antioxidant,secretory immunoglobulin A(s Ig A)content,apoptosis,gene and protein expression analysis;cecal contents were also collected for microbiota analysis based on 16S r DNA sequencing.Results Dietary EPS promoted the average daily gain(ADG)of broilers during 3–4 weeks of HS(P<0.05).At the end of HS on broilers,the activity of total superoxide dismutase(T-SOD),glutathione S-transferase(GST),and the content of s Ig A in jejunum were improved by EPS supplementation(P<0.05).Besides,dietary EPS reduced the epithelial cell apoptosis of jejunum and ileum in heat-stressed broilers(P<0.05).Addition of EPS in HS group broilers'diet upregulated the relative m RNA expression of Occludin,ZO-1,γ-GCLc and IL-10 of the jejunum(P<0.05),whereas downregulated the relative m RNA expression of NF-κB p65,TNF-αand IL-1βof the jejunum(P<0.05).Dietary EPS increased the protein expression of Occludin and ZO-1,whereas it reduced the protein expression of NF-κB p65 and MLCK(P<0.01)and tended to decrease the protein expression of TNF-α(P=0.094)in heat-stressed broilers.Furthermore,the proportions of Bacteroides and Oscillospira among the three groups were positively associated with jejunal apoptosis and pro-inflammatory cytokine expression(P<0.05)and negatively correlated with jejunal Occludin level(P<0.05).However,the proportions of Lactobacillus,Barnesiella,Subdoligranulum,Megasphaera,Collinsella,and Blautia among the three groups were positively related to ADG(P<0.05).Conclusions EPS can be used as a feed additive in yellow-feathered broilers.It effectively improves growth performance and alleviates HS-induced intestinal injury by relieving inflammatory damage and improving the tight junction proteins expression.These beneficial effects may be related to inhibiting NF-κB/MLCK signaling pathway activation and regulation of cecal microbiota.

Effect of organic mineral supplementation in reducing oxidative stress in Holstein calves during short‑term heat stress and recovery conditions

Background This study investigated the effects of inorganic and organic minerals on physiological responses,oxidative stress reduction,and rumen microbiota in Holstein bull calves(123.81±9.76 kg;5 months old)during short-term heat stress(HS)and recovery periods.Eight Holstein calves were randomly assigned to four treatment groups:no mineral supplementation(Con),inorganic minerals(IM),organic minerals(OM),and high-concentration organic minerals(HOM)and two thermal environments(HS and recovery)using 4×2 factorial arrangement in a crossover design of four periods of 35 d.Calves were maintained in a temperature-controlled barn.The experimental period consisted of 14 d of HS,14 d of recovery condititon,and a 7-d washing period.Results Body temperature and respiration rate were higher in HS than in the recovery conditions(P<0.05).Selenium concentration in serum was high in the HOM-supplemented calves in both HS(90.38μg/dL)and recovery periods(102.00μg/dL)(P<0.05).During the HS period,the serum cortisol was 20.26 ng/mL in the HOM group,which was 5.60 ng/mL lower than in the control group(P<0.05).The total antioxidant status was the highest in the OM group(2.71 mmol Trolox equivalent/L),followed by the HOM group during HS,whereas it was highest in the HOM group(2.58 mmol Trolox equivalent/L)during the recovery period(P<0.05).Plasma malondialdehyde and HSP70 levels were decreased by HOM supplementation during the HS and recovery periods,whereas SOD and GPX levels were not significantly affected(P>0.05).The principal coordinate analysis represented that the overall rumen microbiota was not influenced by mineral supplementation;however,temperature-induced microbial structure shifts were indicated(PERMANOVA:P<0.05).At the phylum level,Firmicutes and Actinobacteria decreased,whereas Fibrobacteres,Spirochaetes,and Tenericutes increased(P<0.05),under HS conditions.The genus Treponema increased under HS conditions,while Christensenella was higher in recovery conditions(P<0.05).Conclusion HOM supplementation during HS reduced cortisol concentrations and increased total antioxidant status in Holstein bull calves,suggesting that high organic mineral supplementation may alleviate the adverse effects of HS.

Differential expression of miRNAs and mRNAs in the livers of largemouth bass Micropterus salmoides under heat stress

Global warming threatens freshwater ecosystems and compromises fish survival.To elucidate the role of miRNAs in the livers of heat stressed largemouth bass,juvenile fish was subject to heat stress under 37°C.Both mRNA-seq and miRNA-seq were conducted on the liver tissues under control and heat stress conditions.Differential gene expression analysis and enrichment analysis were performed on mRNA and miRNA expression profiles.A total of 406 differentially expressed genes(DEGs)were discovered,of which 212 were up-regulated and 194 were down-regulated.Most of the DEGs were significantly implicated in the regulation of“protein processing in endoplasmic reticulum”,“proteasome”,“steroid biosynthesis”,and“ornithine decarboxylase inhibitor activity”pathways.In addition,47 differentially expressed miRNAs(DEMs)were identified in largemouth bass livers under heat stress,including 21 up-regulated and 25 down-regulated miRNAs.A negatively regulated miRNA-mRNA network including 12 miRNAs and 19 mRNAs was constructed with DEMs involved in“protein degradation”,“calcium ion regulation”,“cell apoptosis”,and“lipid metabolism”.Moreover,this study indicated novel-miR-144 activated the IRE1 signaling pathway by targeting txndc5 to induce liver apoptosis in largemouth bass under heat stress.This study revealed the involvement of miRNA regulation in largemouth bass in response to heat stress.

Impact of Dietary Lactobacillus plantarum Postbiotics on the Performance of Layer Hens under Heat Stress Conditions

This experiment was conducted to determine the performance of heat-stressed layers fed a diet containing the probiotic Lactobacillus plantarum RS5 or its products of fermentation (postbiotics). Twenty-week-old Isa White layers, were subdivided into six treatments of 32 individually caged birds. Half of the birds were reared under regular temperature conditions, while the other half was subjected to cyclic daily heat stress. Layers were offered one of three diets: 1) Control;2) Control + Lactobacillus plantarum RS5 probiotic;3) Control + Lactobacillus plantarum RS5 postbiotics. Birds were tested for performance and visceral organ development for 5 months. Heat stress negatively affected the birds’ feed intake, egg weight, shell weight percentage, Haugh unit, shell thickness, yolk color, body weight and spleen weight percentage. Postbiotics significantly increased egg production (p < 0.05) in comparison to the control and the probiotic fed group (94.8% vs 92.6% vs 93.1%, respectively). Birds under probiotic or postbiotic diet showed a significantly higher (p < 0.05) feed intake and egg weight, although the probiotic had a more pronounced and gradual effect. Specific gravity, yolk weight percentage and shell thickness didn’t show differences among dietary groups. The Haugh Unit was significantly higher (p < 0.05) in probiotic group which also showed a significantly lower yolk color index (p < 0.05). The different feed treatments did not impact the bird’s viscera weight percentage, except for the ileum that was significantly lower (p < 0.05) under postbiotic supplementation. Both probiotics and postbiotics could be used as a potential growth promoters and might alleviate heat stress impact in poultry industry.

Heat stress in pigs and broilers:role of gut dysbiosis in the impairment of the gut-liver axis and restoration of these effects by probiotics,prebiotics and synbiotics

Heat stress is one of the most challenging stressors for animal production due to high economic losses resulting from impaired animal’s productivity,health and welfare.Despite the fact that all farm animal species are susceptible to heat stress,birds and pigs are particularly sensitive to heat stress due to either lacking or non-functional sweat glands.Con-vincing evidence in the literature exists that gut dysbiosis,a term used to describe a perturbation of commensal gut microbiota,develops in broilers and pigs under heat stress.Owing to the protective role of commensal bacteria for the gut barrier,gut dysbiosis causes a disruption of the gut barrier leading to endotoxemia,which contributes to the typical characteristics of heat stressed broilers and growing and growing-finishing pigs,such as reduced feed intake,decreased growth and reduced lean carcass weight.A substantial number of studies have shown that feeding of probiotics,prebiotics and synbiotics is an efficacious strategy to protect broilers from heat stress-induced gut barrier disruption through altering the gut microbiota and promoting all decisive structural,biochemical,and immunologi-cal elements of the intestinal barrier.In most of the available studies in heat stressed broilers,the alterations of gut microbiota and improvements of gut barrier function induced by feeding of either probiotics,prebiotics or synbiot-ics were accompanied by an improved productivity,health and/or welfare when compared to non-supplemented broilers exposed to heat stress.These findings indicate that the restoration of gut homeostasis and function is a key target for dietary interventions aiming to provide at least partial protection of broilers from the detrimental impact of heat stress conditions.Despite the fact that the number of studies dealing with the same feeding strategy in heat stressed pigs is limited,the available few studies suggest that feeding of probiotics might also be a suitable approach to enhance productivity,health and welfare in pigs kept under heat stress conditions.

Chronic heat stress induces renal fibrosis and mitochondrial dysfunction in laying hens

Background Heat stress in laying hens negatively affects egg production and shell quality by disrupting the homeo-stasis of plasma calcium and phosphorus levels.Although the kidney plays an important role in calcium and phos-phorus homeostasis,evidence regarding the effect of heat stress on renal injury in laying hens is yet to be elucidated.Therefore,the aim of this study was to evaluate the effects of chronic heat stress on renal damage in hens during laying periods.Methods A total of 16 white-leghorn laying hens(32 weeks old)were randomly assigned to two groups(n=8).One group was exposed to chronic heat stress(33°C for 4 weeks),whereas the other group was maintained at 24°C.Results Chronic heat exposure significantly increased plasma creatinine and decreased plasma albumin levels(P<0.05).Heat exposure also increased renal fibrosis and the transcription levels of fibrosis-related genes(COLA1A1,αSMA,and TGF-β)in the kidney.These results suggest that renal failure and fibrosis were induced by chronic heat exposure in laying hens.In addition,chronic heat exposure decreased ATP levels and mitochondrial DNA copy number(mtDNA-CN)in renal tissue,suggesting that renal mitochondrial dysfunction occurs under conditions of heat stress.Damaged mitochondria leak mtDNAs into the cytosol and mtDNA leakage may activate the cyclic GMP-AMP synthase(cGAS)stimulator of interferon genes(STING)signaling pathway.Our results showed that chronic heat exposure activated the cGAS-STING pathway as indicated by increased expression of MDA5,STING,IRF7,MAVS,and NF-κB levels.Furthermore,the expression of pro-inflammatory cytokines(IL-12)and chemokines(CCL4 and CCL20)was upregulated in heat-stressed hens.Conclusions These results suggest that chronic heat exposure induces renal fibrosis and mitochondrial damage in laying hens.Mitochondrial damage by heat stress may activate the mtDNA-cGAS-STING signaling and cause subse-quent inflammation,which contributes to the progression of renal fibrosis and dysfunction.

Heat stress affects dairy cow health status through blood oxygen availability

Background Rises in global warming and extreme weather occurrence make the risk of heat stress(HS)induced by high ambient temperatures more likely in high-yielding dairy cows,resulting in low milk quality and yield.In ani-mals,oxygen is involved in many physiological and metabolic processes,but the effects of HS on oxygen metabolism remain unclear.Thus,the current study aimed to investigate how oxygen metabolism plays a role in health status of dairy cows by measuring the milk yield,milk composition,and blood biochemical variables of cows under different levels of HS:none(No-HS),mild(Mild-HS),and moderate HS(Mod-HS).Results The HS significantly increased rectal temperature(Ptreat<0.01)and respiration rate(Ptreat<0.01).Under Mod-HS,greater Na+(P<0.05)and lower total CO_(2),and pH(P<0.05)were observed relative to those under No-HS and Mild-HS.Oxygen concentrations in both coccygeal artery and mammary vein(Ptreat<0.01)were lower under Mod-HS than under No-HS.Coccygeal vein concentrations of heat shock protein 90(HSP90)(P<0.05)increased during Mod-HS compared with those in cows under No-HS.Malondialdehyde increased during Mod-HS,and glu-tathione peroxidase(P<0.01)increased during Mild-HS.Coccygeal vein concentrations of vascular endothelial growth factor(P<0.01),heme oxygenase-1(P<0.01),and hypoxia-inducible factor 1α(P<0.01)were greater in cows under Mod-HS than those under No-HS.Red blood cell count(P<0.01)and hemoglobin concentration(P<0.01)were lower in the coccygeal vein of dairy cows under Mild-and Mod-HS than those of cows under No-HS.Conclusions Exposure to HS negatively impacts the health status and lactation performance of dairy cows by limit-ing oxygen metabolism and transportation.However,the specific mechanism by which HS affects mammary function in cows remains unclear and requires further exploration.

Analysis of differentially expressed genes in the sea cucumber Apostichopus japonicus under heat stress

The sea cucumber Apostichopus japonicus plays important roles in marine benthic ecosystem as environmental cleaners,and it is the important aquaculture species in China.High water temperature poses critical threat for the survival of A.japonicus,which has resulted in extensive death in summer.To explore the genes expression profiles under different levels of heat stress,the high-throughput RNA-seq was applied in this study.Our results revealed a total of 1371,1225 and 1408 differentially expressed genes(DEGs)in 26℃for 6 h,26℃for 48 h and 30℃for 6 h respectively in comparison with Control group.The pathway analysis suggested“Protein processing in endoplasmic reticulum(ER)”was significantly enriched in all these heat stress(HS)treatment groups.The expression results of key DEGs in this pathway(Hsp70,Derlin,NEF,PDI,GPR94 and ERP57)by qRT-PCR was in accordance with the RNA-seq data.The subcluster analysis of DEGs revealed that a variety of heat shock proteins(HSPs)and calcium ion binding proteins had an obvious up-regulated expression in 26℃for 6 h,comparatively low expression in 26℃for 48 h,and the highest expression in 30℃for 6 h.The other DEGs subcluster,consisting of critical components of extracellular matrix(ECM)and a subset of peptidases and proteases,showed significantly rising tendency in 30℃for 6 h.Additionally,the expression of matrix metalloproteases(MMP1,MMP16 and MMP19)was prominently affected by HS,and peaked in 30℃for 6 h.This study provides a series of candidate genes for further study about heat shock response in A.japonicus,especially genes associated with protein processing in ER and regulation of ECM,which also offers new insights into cellular homeostasis under stressful conditions in marine invertebrates.

Transcriptome analysis reveals the mechanism of chronic heat stress on meat quality of broilers

Background:Chronic heat stress has a negative impact on poultry meat quality.Although this has been extensively investigated,previous studies have primarily focused on metabolic alterations and oxidative stress in the pectoralis major(PM)muscle under chronic heat stress,and not all of the underlying molecular mechanisms are completely understood.Methods:A total of 144 male Arbor Acres broilers(28 d old)were randomly allocated into 3 treatment groups:(1)the normal control(NC)group,with broilers raised at 22℃and fed a basal diet;(2)the heat stress(HS)group,with birds raised at 32℃and fed a basal diet;and(3)the pair-fed(PF)group,with birds raised at 22℃and fed the amount of feed equal to the feed consumed on the previous day by the HS group.The experiment lasted for 14 d.Results:Chronic heat stress decreased the average daily feed intake and average daily gain,increased feed:gain ratio(P<0.05);and increased drip loss,cooking loss,shear force,hardness,and decreased p H,redness(a*);and springiness of PM muscle(P<0.05).Furthermore,chronic heat stress decreased muscle fiber density,increased connective tissue,and led to intracellular vacuolation.The transcriptome analyses indicated that the effect of chronic heat stress on meat quality was not only related to metabolism and oxidative stress,but also to signal transduction,immune system,transport and catabolism,cell growth and death,and muscle structure.Conclusions:Chronic heat stress has a negative impact on the growth performance,meat quality,and the PM muscle structure of broilers.Transcriptome analysis revealed a comprehensive understanding of the mechanism of the chronic heat stress-induced deterioration of broiler meat quality at the transcriptional level.

Effects of Heat Stress during Seed Filling Stage on Brassica napus Seed Oil Accumulation and Chlorophyll Fluorescence Characteristics

As global temperature rise,the threat of heat stress to rapeseed production is becoming more obvious.Exploring the response characteristics of two important biological pathways,oil accumulation and photosynthesis,to heat stress during B.napus seed filling is helpful in the genetic improvement of heat-tolerant rapeseed.The effects of heat stress on seed oil accumulation and chlorophyll fluorescence characteristics of 29 B.napus germplasms with different oil content and environmental sensitivity,including 6 rapeseed varieties which exhibited environmentsensitive/insensitive and with high,medium or low oil content,were tested by whole plant heat stress or the in vitro silique culture system.Both assay exhibited similar trend on oil content of the rapeseed germplasms.The heat effect on the chlorophyll fluorescence kinetic parameters F_(v)/F_(m),ETR and Y(Ⅱ)were also consistent.Heat stress significantly decreased oil content,although there was abundant genetic variation on heat tolerance among the genotypes.Correlation analysis showed that the decrease rate of F_(v)/F_(m) of silique heat-stressed B.napus developing seed was positive correlative to the decrease rate of mature seed oil content of the whole plant heat-stressed rapeseed(R=0.9214,P-value<0.01).Overall,the results indicated that heat stress inhibited oil accumulation and photosynthesis in B.napus developing seed.The decrease rate of chlorophyll fluorescence parameter F_(v)/F_(m) of heat-stressed developing seed could be used as the index of heat tolerant rapeseed identification.Further,two heat insensitive rapeseed varieties with high oil content were identified.

Performance of Dairy Cows Supplemented with By-Pass Fat under Heat Stress Conditions

The objective of this study was to determine the effect of supplementation with a protected fat source on the productive response, metabolic environment and physiological indicators in Holstein cows under heat stress conditions during a 12-week experimental period. Thirty Holstein cows were distributed in 15 blocks by parity (2.0 ± 1.1), days in milk (182 ± 80) and milk production (29.4 ± 5.7 kg·day-1) at the beginning of the trial and randomly assigned within each block to the following treatments (diets): SPF: supplementation with protected fat or WPF: without supplementation with protected fat. All the cows were kept in a dry-lot where they were given a partial mixed ration (PMR) ad libitum while in the milking parlor they received individual supplementation depending on the treatment. The SPF diet contained 4.0 kg·day-1 concentrate in pellet form + 0.6 kg·day-1 ground corn grain + 0.7 kg·day-1 protected fat, while the WPF diet was similar to that offered in SPF, but the protected fat was isoenergetically replaced by ground corn grain. The fat supplement contained fats of animal and vegetable origin and microencapsulation was used for its preparation. Total dry matter and metabolic energy intakes were similar (p > 0.05) between treatments. Fat corrected milk (4% FCM) production was higher (p = 0.04), while energy corrected milk and fat productions tended (p = 0.06) to be higher in cows from the SPF group, without effects (p > 0.05) on the rest of the milk production and composition parameters. These results could be attributed to an improvement in the efficiency of the use of the energy consumed. Protected fat supplementation neither modified the metabolic profile, nor reduced the respiratory rate and body temperature of heat-stressed cows. Future research is needed to explain this latter result.

Preliminary Studies on the Relationship betweenAutoantibodies to Heat Stress Proteins and Heat Injury of Pilots during Acute Heat Stress

Comparison in the heart rate, oral temperature and lymphocyte DNAdamage during heat stress was made in pilots with negative antibodies to heatstress proteins (HSPs) and those with positive antibodies in the man-made climate room with Western blot and comet assay. Our results showed that the increase in oral temperature, heart rate and lymphocyte DNA damage in pilots with the posi-tive antibodies to HSPs were higher than those in pilots with the negative antibod-ies during heat stress. These results indicated that the presence of autoantibodies in plasma of pilots might reflect heat damage and high sensitivity to heat.

Weakened carbon and nitrogen metabolisms under post-silking heat stress reduce the yield and dry matter accumulation in waxy maize

Post-silking high temperature is one of the abiotic factors that affects waxy maize(Zea mays L. sinensis Kulesh) growth in southern China. We conducted a pot trial in 2016–2017 to study the effects of post-silking daytime heat stress(35°C) on the activities of enzymes involved in leaf carbon and nitrogen metabolisms and leaf reactive oxygen species(ROS) and water contents. This study could improve our understanding on dry matter accumulation and translocation and grain yield production. Results indicated that decreased grain number and weight under heat stress led to yield loss, which decreased by 20.8 and 20.0% in 2016 and 2017, respectively. High temperature reduced post-silking dry matter accumulation(16.1 and 29.5% in 2016 and 2017, respectively) and promoted translocation of pre-silking photoassimilates stored in vegetative organs, especially in leaf. The lower leaf water content and chlorophyll SPAD value, and higher ROS(H2O2 and O2^-·) content under heat stress conditions indicated accelerated senescent rate. The weak activities of phosphoenolpyruvate carboxylase(PEPCase), Ribulose-1,5-bisphosphate carboxylase(Ru BPCase), nitrate reductase(NR), and glutamine synthase(GS) indicated that leaf carbon and nitrogen metabolisms were suppressed when the plants suffered from a high temperature during grain filling. Correlation analysis results indicated that the reduced grain yield was mainly caused by the decreased leaf water content, weakened NR activity, and increased H2O2 content. The increased accumulation of grain weight and post-silking dry matter and the reduced translocation amount in leaf was mainly due to the increased chlorophyll SPAD value and NR activity. Reduced PEPCase and Ru BPCase activities did not affect dry matter accumulation and translocation and grain yield. In conclusion, post-silking heat stress down-regulated the leaf NR and GS activities, increased the leafwater loss rate, increased ROS generation, and induced pre-silking carbohydrate translocation. However, it reduced the post-silking direct photoassimilate deposition, ultimately, leading to grain yield loss.

Foliar application of sodium hydrosulfide(NaHS),a hydrogen sulfide(H2S) donor,can protect seedlings against heat stress in wheat(Triticum aestivum L.)

Temperature extremes represent an important limiting factor to plant growth and productivity. Low concentration of hydrogen sulfide （H2S） has been proven to function in physiological responses to various stresses. The present study evaluated the effect of foliar application of wheat seedlings with a H2S donor, sodium hydrosulfide （NariS）, on the response to acute heat stress. The results showed that pretreatment with NariS could promote heat tolerance of wheat seedlings in a dose-depen- dent manner. Again, it was verified that H2S, rather than other sulfur-containing components or sodion derived from NariS solution, should contribute to the positive role in promoting wheat seedlings against heat stress. To further study antioxidant mechanisms of NariS-induced heat tolerance, superoxide dismutase （SOD, EC 1.15.1.1 ）, catalase （CAT, EC 1.11.1.6） and ascorbate peroxidase （APX, EC 1.11.1.11 ） activities, and HzS, hydrogen peroxide （H2O2）, malonaldehyde （MDA）, and soluble sugar contents in wheat seedlings were determined. The results showed that, under heat stress, the activities of SOD, CAT, and APX, H2S, H2O2, MDA, and soluble sugar contents in NaHS-pretreated seedlings and its control all increased. Meanwhile, NaHS-pretreated seedlings showed higher antioxidant enzymes activities and gene expression levels as well as the H2S and soluble sugar levels, and lower H2O2, MDA contents induced by heat stress. While little effect was detected in antioxidant enzymes activities and soluble substances contents in pretreated wheat seedlings compared with its control under normal culture conditions （data not shown）. All of our results suggested that exogenous NariS could alleviate oxidative damage and improve heat tolerance by regulating the antioxidant system in wheat seedlings under heat stress.

Heat Stress Upregulates the Expression of TLR4 and Its Alternative Splicing Variant in Bama Miniature Pigs

Alternative splicing is a cellular mechanism in eukaryotes that results in considerable diversity ofgene products. It plays an important role in several diseases and cellular signal regulation. Heat stress is a major factor that induces immunosuppression in pigs. Little is known about the correlation between alternative splicing and heat stress in pigs. Therefore, this study aimed to clone, sequence and quantify the alternative splicing variant of toll-like receptor 4 （TLR4） in Bama miniature pigs （Sus scrofa domestica） following exposure to heat stress. The results showed that the second exon of TLR4 was spliced and 167 bp shorter in the alternative splicing variant, and the protein was putatively identified as a type of truncated membrane protein consisting of extramembrane, transmembrane and intramembrane regions lacking a signal peptide. Further, it was not a non- classical secretory protein. Five potential reference genes were screened for their potential as reliable standards to quantify the expression of TLR4 alternative spliced variants by real-time quantitative reverse-transcriptase polymerase chain reaction （qRT-PCR）. The stability of these reference genes was ranked using the geNorm and NormFinder programs, and ribosomal protein L4 （RPL4） and TATA box-binding protein （TBP） were found to be the two genes showing the most stable expression in the in vitro cultured peripheral blood mononuclear ceils （PBMCs） during heat shock. The mRNA level of the TLR4 gene （both classical and spliced） in stressed pigs increased significantly （P〈0.05）. Further, the expression levels of the alternative spliced variant of TLR4 （TLR4-ASV） showed a 2-3 folds increase in heat-stressed PBMCs as compared to control pigs. The results of the present study suggested that heat shock might modulate the host immune response by regulating the expressions of TLR4 and its alternative splicing variant.

Review of the impact of heat stress on reproductive performance of sheep

Heat stress significantly impairs reproduction of sheep,and under current climatic conditions is a significant risk to the efficiency of the meat and wool production,with the impact increasing as global temperatures rise.Evidence from field studies and studies conducted using environmental chambers demonstrate the effects of hot temperatures(≥32℃)on components of ewe fertility(oestrus,fertilisation,embryo survival and lambing)are most destructive when experienced from 5 d before until 5 d after oestrus.Temperature controlled studies also demonstrate that ram fertility,as measured by rates of fertilisation and embryo survival,is reduced when mating occurs during the period 14 to 50 d post-heating.However,the contribution of the ram to heat induced reductions in flock fertility is difficult to determine accurately.Based primarily on temperature controlled studies,it is clear that sustained exposure to high temperatures(≥32℃)during pregnancy reduces lamb birthweight and will,therefore,decrease lamb survival under field conditions.It is concluded that both ewe and ram reproduction is affected by relatively modest levels of heat stress(≥32℃)and this is a concern given that a significant proportion of the global sheep population experiences heat stress of this magnitude around mating and during pregnancy.Despite this,strategies to limit the impacts of the climate on the homeothermy,behaviour,resource use and reproduction of extensively grazed sheep are limited,and there is an urgency to improve knowledge and to develop husbandry practices to limit these impacts.

Physiological responses of artificial moss biocrusts to dehydration-rehydration process and heat stress on the Loess Plateau, China

Ex-situ cultivation of biological soil crusts （biocrusts） is a promising technology to produce materials that can induce the recovery of biocrusts in the field for the purposes of preventing soil erosion and improving hydrological function in degraded ecosystems. However, the ability of artificially cultivated biocrusts to survive under adverse field conditions, including drought and heat stresses, is still relatively unknown. Mosses can bolster biocrust resistance to the stresses （e.g., drought and heat） and the resistance may be introduced prior to field cultivation. In this study, we subjected the well-developed artificial moss biocrusts （dominant species of Didjmodon vinealis （Brid.） Zand.） that we cultivated in the phytotron to a dehydration-rehydration experiment and also a heat stress experiment and measured the activities of protective enzymes （including peroxidase （POD）, superoxide dismutase （SOD）, and catalase （CAT）） and the contents of osmoregulatory substances （including soluble proteins and soluble sugars） and malondialdehyde （MDA, an indicator of oxidative stress） in the stem and leaf fragments of mosses. The results showed that, during the dehydration process, the activities of protective enzymes and the contents of osmoregulatory substances and MDA gradually increased with increasing duration of drought stress （over 13 days）. During the rehydration process, values of these parameters decreased rapidly after 1 d of rehydration. The values then showed a gradual decrease for 5 days, approaching to the control levels. Under heat stress （45℃）, the activities of protective enzymes and the content of soluble proteins increased rapidly within 2 h of heat exposure and then decreased gradually with increasing duration of heat exposure. In contrast, the contents of soluble sugars and MDA always increased gradually with increasing duration of heat exposure. This study indicates that artificial moss biocrusts possess a strong drought resistance and this resistance can be enhanced after a gradual dehydration treatment. This study also indicates that artificial moss biocrusts can only resist short-term heat stress （not long-term heat stress）. These findings suggest that short-term heat stress or prolonged drought stress could be used to elevate the resistance of artificial moss biocrusts to adverse conditions prior to field reintroduction.

Functional roles of taurine,L-theanine,Lcitrulline,and betaine during heat stress in poultry

Heat stress(HS)is an important environmental stress factor affecting poultry production on a global scale.With the rise in ambient temperature and increasing effects of global warming,it becomes pertinent to understand the effects of HS on poultry production and the strategies that can be adopted to mitigate its detrimental impacts on the performance,health,welfare,immunity,and survival of birds.Amino acids(AAs)have been increasingly adopted as nutritional modifiers in animals to ameliorate the adverse effects of HS.They are essential for protein synthesis,growth,maintenance,reproduction,immunity,stress response,and whole-body homeostasis.However,HS tends to adversely affect the availability,transport,absorption,and utilization of these AAs.Studies have investigated the provision of these AAs to poultry during HS conditions,and variable findings have been reported.Taurine,L-theanine,and L-citrulline are non-essential amino acids that are increasingly gaining attention as nutritional supplements in HS animals.Similarly,betaine is an amino acid derivative that possesses favorable biological properties which contributes to its role as a functional additive during HS.Of particular note,taurine is negligible in plants,while betaine,L-theanine,and L-citrulline can be found in selected plants.These nutrients are barely found in feed ingredients,but their supply has been shown to elicit important physiological roles including anti-stress effects,anti-oxidative,anti-inflammatory,gut promoting,and immunomodulatory functions.The present review provides information on the use of these nutritionally and physiologically beneficial nutrients as functional additives to poultry diets during HS conditions.Presently,although several studies have reported on the positive effects of these additives in human and murine studies,however,there is limited information regarding their utilization during heat stress in poultry nutrition.Therefore,this review aims to expound on the functional properties of these nutrients,their potentials for HS alleviation,and to stimulate further researches on their biological roles in poultry nutrition.

Application of moderate nitrogen levels alleviates yield loss and grain quality deterioration caused by post-silking heat stress in fresh waxy maize

High temperature(HT)during grain filling is one of the most important environmental factors limiting maize yield and grain quality.Nitrogen(N)fertilizer is essential for maintaining normal plant growth and defense against environmental stresses.The effects of three N rates and two temperature regimes on the grain yield and quality of fresh waxy maize were studied using the hybrids Suyunuo 5(SYN5)and Yunuo 7(YN7)as materials.N application rates were 1.5,4.5,and 7.5 g plant-1,representing low,moderate,and high N levels(LN,MN,and HN,respectively).Mean day/night temperatures during the grain filling of spring-and summer-sown plants were 27.6/21.0°C and 28.6/20.0°C for ambient temperature(AT)and 35/21.0°C and 35/20.0°C for HT,respectively.On average,HT reduced kernel number,weight,yield,and moisture content by 29.8%,17.9%,38.7%,and 3.3%,respectively.Kernel number,weight,yield,moisture,and starch contents were highest under MN among the three N rates under both temperature regimes.HT reduced grain starch content at all N levels.HT increased grain protein content,which gradually increased with N rate.Mean starch granule size under MN was larger(10.9μm)than that under LN and HN(both 10.4μm)at AT.However,the mean size of starch granules was higher under LN(11.7μm)and lower under MN(11.2μm)at HT.Iodine binding capacity(IBC)was lowest under MN and highest under HN among the three N levels under both temperature regimes.In general,IBC at all N rates was increased by HT.Peak viscosity(PV)was gradually reduced with increasing N rate at AT.In comparison with LN,PV was increased by MN and decreased by HN at HT.Retrogradation percentage gradually increased with N rate at AT,but was lowest under MN among the three N rates at HT.LN+AT and MN+HT produced grain with high pasting viscosity and low retrogradation tendency.MN application could alleviate the negative effects of HT on the grain yield and quality of fresh waxy maize.

Relationship Between Heart Damages and HSPs mRNA in Persistent Heat Stressed Broilers

The relationship between myocardial cell damages and HSPs mRNA transcription in heat stressed broilers was studied using a spectrophotometer, the histopathological technique, and fluorescence quantitative reverse transcription PCR （FQ RT-PCR）. The results showed that the activities of creatine kinase （CK） and glutamic-pyruvic transaminase （GPT） were induction during the persistent heat stress. The major lesions of the myocardial fibers were granular degeneration and necrosis. The transcription of constitutive or cognate heat shock protein 70 （HSC70） mRNA was changeable. The transcription of heat shock protein 70 （HSPT0） mRNA was increased obviously in the course of persistent heat stress. The results showed that the change of HSC70 mRNA transcription was contrary to the activity of CK, and the level of HSC70 mRNA transcription must be used as a symbol of the myocardial cell damages in the course of persistent heat stress.