Transcriptome analysis reveals steroid hormones biosynthesis pathway involved in abdominal fat deposition in broilers

Excessive abdominal fat deposition reduces the feed efficiency and increase the cost of production in broilers.Therefore,it is an important task for poultry breeders to breed broilers with low abdominal fat.Abdominal fat deposition is a highly complex biological process,and its molecular basis remains elusive.In this study,we performed transcriptome analysis to compare gene expression profiles at different stages of abdominal fat deposition to identify the key genes and pathways involved in abdominal fat accumulation.We found that abdominal fat weight(AFW)increased gradually from day 35(D35)to 91(D91),and then decreased at day 119(D119).Accordingly,after detecting differentially expressed genes(DEGs)by comparing gene expression profiles at D35 vs.D63 and D35 vs.D91,and identifying gene modules associated with fat deposition by weighted gene co-expression network analysis(WGCNA),we performed intersection analysis of the detected DEGs and WGCNA gene modules and identified 394 and 435 intersecting genes,respectively.The results of the Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses showed that the steroid hormone biosynthesis and insulin signaling pathways were co-enriched in all intersecting genes,steroid hormones have been shown that regulated insulin signaling pathway,indicating the importance of the steroid hormone biosynthesis pathway in the development of broiler abdominal fat.We then identified 6 hub genes(ACTB,SOX9,RHOBTB2,PDLIM3,NEDD9,and DOCK4)related to abdominal fat deposition.Further analysis also revealed that there were direct interactions between 6 hub genes.SOX9 has been shown to bind to proteins required for steroid hormone receptor binding,and RHOBTB2 indirectly regulates the steroid hormones biosynthesis through cyclin factor,and ultimately affect fat deposition.Our results suggest that the genes RHOBTB2 and SOX9 play an important role in fat deposition in broilers,by regulating steroid hormone synthesis.These findings provide new targets and directions for further studies on the mechanisms of fat deposition in chicken.

Abdominal Fat Accumulation with Hyperuricemia and Hypercholesterolemia Quail Model Induced by High Fat Diet

Objective To establish abdominal fat accumulation with hyperuricemia and hypercholesterolemia quail model fed with high fat diet. And then to investigate the pathological characteristics of this quail model. Methods Thirty Longcheng quails were randomly divided into two groups： control group and model group （n=15）. The control group quails were fed with normal diet and model group quails were fed with high fat diet for 14 days. After a 12-hour overnight fast, liver and abdominal fat at euthanasia as well as serum were collected. The levels of serum uric acid, total cholesterol, high density lipoprotein cholesterol （HDL-C）, low density lipoprotein cholesterol （LDL-C）, triglyceride, free fatty acid （FFA）, and blood glucose were assayed. The activity changes of adenosine deaminase （ADA）, xanthine oxidase （XOD）, lipoprotein lipase （LPL）, hepatic lipase （HL）, and fatty acid synthetase （FAS） were analyzed. Results Compared with control group, the abdominal fat content （0.74±0.63 vs. 1.36±0.65 g, P〈0.05） and abdominal fat index （0.44%±0.30% vs. 0.85%±0.30%, P〈0.01） as well as live lipid index （3.61%±0.65% vs. 11.33%±2.14%, P〈0.01） in model group significantly increased; the levels of serum uric acid （210.61±94,76 vs. 304.25±141.94 /amol/L, P〈0.05）, total cholesterol （4.20±0.51 vs. 20.10±11.25 mmol/L, P〈0.01）, LDL-C （1.16±0.29 vs. 10.78±6.48 mmol/L, P〈0.01）, and FFA （0.39±0.14 vs. 0.55±0.15 mmol/L, P〈0.01） in model group significantly increased; HDL-C （5.85±0.95 vs. 4.14±2.03 mmol/L, P〈0.05） significantly decreased; the levels of triglyceride and blood glucose had no significant changes （P〉0.05）; the activities of ADA （9.71±3.05 vs. 17.19±5.10 U/ml, P〈0.01） and XOD （10.58±6,88 vs. 19.22＋9.44 U/L, P〈0.01） in model group significantly increased; and FAS, LPL, HL had no significant changes （P〉0.05）. Conclusions High fat diet can induce abdominal fat accumulation with hyperuricemia and hypercholesterolemia quail model. The changes of uric acid and lipid metabolic enzyme activities may he the pathological mechanism of abdominal fat accumulation with hyperuricemia and hypercholesterolemia.

Correlation of fatty liver and abdominal fat distribution using a simple fat computed tomography protocol

AIM: To evaluate the relationship between hepatic fat infiltration and abdominal fat volume by using computed tomography (CT).METHODS: Three hundred and six patients who visited our obesity clinic between November 2007 and April 2008 underwent fat protocol CT scans.The age range of the patients was 19 to 79 years and the mean age was 49 years.The male to female ratio was 116:190.Liver and spleen attenuation measurements were taken with three regions of interests (ROIs) from the liver and two ROIs from the spleen.Hepatic attenuation indices (HAIs) were measured as follows: (1) hepatic parenchymal attenuation (CTLP);(2) liver to spleen attenuation ratio (LS ratio);and (3) difference between hepatic and splenic attenuation (LSdif).Abdominal fat volume was measured using a 3 mm slice CT scan starting at the level of the umbilicus and was automatically calculated by a workstation.Abdominal fat was classified into total fat (TF),visceral fat (VF),and subcutaneous fat (SF).We used a bivariate correlation method to assess the relationship between the three HAIs and TF,VF,and SF.RESULTS: There were significant negative correlations between CTLP,LS ratio,and LSdif with TF,VF,and SF,respectively.The CTLP showed a strong negative correlation with TF and VF (r = -0.415 and -0.434,respectively,P < 0.001).The correlation between CTLP and SF was less significant (r = -0.313,P < 0.001).CONCLUSION: Fatty infiltration of the liver was correlated with amount of abdominal fat and VF was more strongly associated with fatty liver than SF.

Integration of association and computational methods reveals functional variants of LEPR gene for abdominal fat content in chickens

Leptin receptor(LEPR)plays a vital role in obesity in humans and animals.The objective of this study is to assess LEPR functional variants for chicken adipose deposition by integration of association and in-silico analysis using a unique chicken population,the Northeast Agricultural University broiler lines divergently selected for abdominal fat content(NEAUHLF).Five online bioinformatics tools were used to predict the functionality of the single nucleotide polymorphisms(SNPs)in coding region.Further,the possible structure–function relationship of high confidence SNPs was determined by bioinformatics analyses,including the conservation and stability analysis based on amino acid residues,prediction of protein ligand-binding sites,and the superposition of protein tertiary structure.Meanwhile,we analyzed the association between abdominal fat traits and 20 polymorphisms of chicken LEPR gene.The integrated results showed that rs731962924(N867I)and rs13684622(C1002R)could lead to striking changes in the structure and function of proteins,of which rs13684622(C1002R)was significantly associated with abdominal fat weight(AFW,P=0.0413)and abdominal fat percentage(AFP,P=0.0260)in chickens.Therefore,we are of the opinion that rs13684622(C1002R)may be an essential functional SNP affecting chicken abdominal fat deposition,and potentially applied to improvement of broiler abdominal fat in molecular marker-assisted selection(MAS)program.Additionally,the coupling of association with computer electronic predictive analysis provides a new avenue to identify important molecular markers for breeders.

Classifying Abdominal Fat Distribution Patterns byUsing Body Measurement Data

This study aims to explore new categorization that characterizes the distribution clusters of visceral and subcutaneous adipose tissues(VAT and SAT)measured by magnetic resonance imaging(MRI),to analyze the relationship between the VAT-SAT distribution patterns and the novel body shape descriptors(BSDs),and to develop a classifier to predict the fat distribution clusters using the BSDs.In the study,66 male and 54 female participants were scanned by MRI and a stereovision body imaging(SBI)to measure participants’abdominal VAT and SAT volumes and the BSDs.A fuzzy c-means algorithm was used to form the inherent grouping clusters of abdominal fat distributions.A support-vector-machine(SVM)classifier,with an embedded feature selection scheme,was employed to determine an optimal subset of the BSDs for predicting internal fat distributions.A fivefold cross-validation procedure was used to prevent over-fitting in the classification.The classification results of the BSDs were compared with those of the traditional anthropometric measurements and the Dual Energy X-Ray Absorptiometry(DXA)measurements.Four clusters were identified for abdominal fat distributions:(1)low VAT and SAT,(2)elevated VAT and SAT,(3)higher SAT,and(4)higher VAT.The cross-validation accuracies of the traditional anthropometric,DXA and BSD measurements were 85.0%,87.5% and 90%,respectively.Compared to the traditional anthropometric and DXA measurements,the BSDs appeared to be effective and efficient in predicting abdominal fat distributions.

Overweight and abdominal fat are associated with normal bone mineral density in patients with ulcerative colitis

BACKGROUND Low bone mineral density(BMD)is common in patients with inflammatory bowel disease.However,nutritional risk factors for low BMD in the ulcerative colitis(UC)population are still poorly understood.AIM To investigate the association of anthropometric indicators and body composition with BMD in patients with UC.METHODS This is a cross-sectional study on adult UC patients of both genders who were followed on an outpatient basis.A control group consisting of healthy volunteers,family members,and close people was also included.The nutritional indicators evaluated were body mass index(BMI),total body mass(TBM),waist circumference(WC),body fat in kg(BFkg),body fat in percentage(BF%),trunk BF(TBF),and also lean mass.Body composition and BMD assessments were performed by dual-energy X-ray absorptiometry.RESULTS The sociodemographic characteristics of patients with UC(n=68)were similar to those of healthy volunteers(n=66)(P>0.05).Most patients(97.0%)were in remission of the disease,58.8%were eutrophic,33.8%were overweight,39.0%had high WC,and 67.6%had excess BF%.However,mean BMI,WC,BFkg,and TBF of UC patients were lower when compared to those of the control group(P<0.05).Reduced BMD was present in 41.2%of patients with UC(38.2%with osteopenia and 2.9%with osteoporosis)and 3.0%in the control group(P<0.001).UC patients with low BMD had lower BMI,TBM,and BFkg values than those with normal BMD(P<0.05).Male patients were more likely to have low BMD(prevalence ratio[PR]=1.86;95%confidence interval[CI]:1.07-3.26).Those with excess weight(PR=0.43;95%CI:0.19-0.97)and high WC(PR=0.44;95%CI:0.21-0.94)were less likely to have low BMD.CONCLUSION Patients with UC in remission have a high prevalence of metabolic bone diseases.Body fat appears to protect against the development of low BMD in these patients.

Visceral Fat Area,Waist Circumference and Metabolic Risk Factors in Abdominally Obese Chinese Adults

Objective To examine the association of visceral adiposity as measured by VFA and WC with lipid and glucose metabolic biomarkers in abdominally obese Chinese adults,and to assess whether WC could be an indicator of visceral fat.Methods WC and VFA were measured in 155 overweight and obese adults.A fasting blood sample was collected from participant（n=118） whose VFA ≥100 cm2 for analyses of lipid and glucose profile.The relationship between VFA and WC and biomarkers was investigated.Results WC and VFA were significantly interrelated.The coincidence rate of abdominal obesity determined by Japanese VFA and Chinese WC criteria increased across age quartiles in women from 51.7% to 96.2%.A large WC was associated significantly with low HDL-cholesterol concentration（P0.01） and the association was weakened by additional control of VFA.WC and VFA were positively associated with glucose,hemoglobin A1c and insulin concentrations（P0.05 except for the association of VFA with insulin：P0.01）,and all the associations were not significant by additional control of either WC or VFA.As WC quartiles increased,significant stepwise increments in triglyceride,glucose,hemoglobin A1c and insulin and descent in HDL-cholesterol were observed.However,triglyceride and HDL-cholesterol were not significantly different when compared across VFA quartiles.Conclusion Higher visceral fat was associated with an adverse lipid and glucose profile.WC can be a moderate predictor for visceral fat and provides a feasible measurement to estimate glucose metabolic risks.Further studies are warranted to establish age-specific WC cutoffs.

CircDOCK7 facilitates the proliferation and adipogenic differentiation of chicken abdominal preadipocytes through the gga‑miR‑301b‑3p/ACSL1 axis

Background Abdominal fat deposition depends on both the proliferation of preadipocytes and their maturation into adipocytes,which is a well-orchestrated multistep process involving many regulatory molecules.Circular RNAs(circRNAs)have emergingly been implicated in mammalian adipogenesis.However,circRNA-mediated regulation in chicken adipogenesis remains unclear.Our previous circRNA sequencing data identified a differentially expressed novel circRNA,8:27,886,180|27,889,657,during the adipogenic differentiation of chicken abdominal preadipocytes.This study aimed to investigate the regulatory role of circDOCK7 in the proliferation and adipogenic differentiation of chicken abdominal preadipocytes,and explore its molecular mechanisms of competing endogenous RNA underlying chicken adipogenesis.Results Our results showed that 8:27,886,180|27,889,657 is an exonic circRNA derived from the head-to-tail splicing of exons 19–22 of the dedicator of cytokinesis 7(DOCK7)gene,abbreviated as circDOCK7.CircDOCK7 is mainly distributed in the cytoplasm of chicken abdominal preadipocytes and is stable because of its RNase R resistance and longer half-life.CircDOCK7 is significantly upregulated in the abdominal fat tissues of fat chickens compared to lean chickens,and its expression gradually increases during the proliferation and adipogenic differentiation of chicken abdominal preadipocytes.Functionally,the gain-and loss-of-function experiments showed that circDOCK7 promoted proliferation,G0/G1-to S-phase progression,and glucose uptake capacity of chicken abdominal preadipocytes,in parallel with adipogenic differentiation characterized by remarkably increased intracellular lipid droplet accumulation and triglyceride and acetyl coenzyme A content in differentiated chicken abdominal preadipocytes.Mechanistically,a pull-down assay and a dual-luciferase reporter assay confirmed that circDOCK7 interacted with gga-miR-301b-3p,which was identified as an inhibitor of chicken abdominal adipogenesis.Moreover,the ACSL1 gene was demonstrated to be a direct target of gga-miR-301b-3p.Chicken ACSL1 protein is localized in the endoplasmic reticulum and mitochondria of chicken abdominal preadipocytes and acts as an adipogenesis accelerator.Rescue experiments showed that circDOCK7 could counteract the inhibitory effects of gga-miR-301b-3p on ACSL1 mRNA abundance as well as the proliferation and adipogenic differentiation of chicken abdominal preadipocytes.Conclusions CircDOCK7 serves as a miRNA sponge that directly sequesters gga-miR-301b-3p away from the ACSL1 gene,thus augmenting adipogenesis in chickens.These findings may elucidate a new regulatory mechanism underlying abdominal fat deposition in chickens.

Effect of resistance training on non-alcoholic fatty-liver disease a randomized-clinical trial

AIM: To evaluate the effect of resistance training (RT) on non alcoholic liver disease (NAFLD) patients.

The Study on Correlation Analysis of Single Nucleotide Polymorphism of IGF2 Gene and Body Fatness Traits in Chicken

Insulin-like growth factor Ⅱhas profound effects on the growth and differentiation of animal embryo. Some researches indicated that it affects the fat metabolism of poultry. This study was designed to investigate the effect of IGF2 on chicken fatness traits. Broiler, Hyline Brown layer and three native breeds (Shiqiza, Beijing You, Baier) were used in this research. Body weight and body composition traits were measured in broiler line at the age of 7 weeks. Primers for exon2 in IGF2 were designed from database of chicken genomic sequence. Polymorphisms were detected by PCR-SSCP and DNA sequencing. The total χ2 test results showed that there was a significant difference (P < 0.01) in the frequency of genotype among breeds. A C/G mutation at base position 139 was found among individuals in broiler line and the least square analysis showed that BB genotype birds had significant lower (P <0.05) abdominal fat weight and percentage of abdominal fat than AA or AB genotype birds. From the results we can conclude putatively that IGF2 gene is the major gene affecting the fatness traits of chicken or it links with the major gene, and the mutation could be used as the molecular genetic marker to select the chicken for low abdominal fat.

Prediction of Abdominal Visceral Obesity From Body Mass Index,Waist Circumference and Waist-hip Ratio in Chinese Adults:Receiver Operating Characteristic Curves Analysis

Objective To evaluate the sensitivity and specificity of body mass index (BMI), waist circumference (WC) and waist-to-hip ratio (WHR) measurements in diagnosing abdominal visceral obesity. Methods BMI, WC, and WHR were assessed in 690 Chinese adults (305 men and 385 women) and compared with magnetic resonance imaging (MRI) measurements of abdominal visceral adipose tissue (VA). Receiver operating characteristic (ROC) curves were generated and used to determine the threshold point for each anthropometric parameter. Results 1) MRI showed that 61.7% of overweight/obese individuals (BMI≥25 kg/m2) and 14.2% of normal weight (BMI<25 kg/m2) individuals had abdominal visceral obesity (VA≥100 cm2). 2) VA was positively correlated with each anthropometric variable, of which WC showed the highest correlation (r=0.73-0.77, P<0.001). 3) The best cut-off points for assessing abdominal visceral obesity were as followed: BMI of 26 kg/m2, WC of 90 cm, and WHR of 0.93, with WC being the most sensitive and specific factor. 4) Among subjects with BMI≥28 kg/m2 or WC≥95 cm, 95% of men and 90% of women appeared to have abdominal visceral obesity. Conclusion Measurements of BMI, WC, and WHR can be used in the prediction of abdominal visceral obesity, of which WC was the one with better accuracy.

Integrative analysis of miRNA and mRNA profiles reveals that gga-miR-106-5p inhibits adipogenesis by targeting the KLF15 gene in chickens

Background:Excessive abdominal fat deposition in commercial broilers presents an obstacle to profitable meat quality,feed utilization,and reproduction.Abdominal fat deposition depends on the proliferation of preadipocytes and their maturation into adipocytes,which involves a cascade of regulatory molecules.Accumulating evidence has shown that microRNAs(miRNAs)serve as post-transcriptional regulators of adipogenic differentiation in mammals.However,the miRNA-mediated molecular mechanisms underlying abdominal fat deposition in chickens are still poorly understood.This study aimed to investigate the biological functions and regulatory mechanism of miRNAs in chicken abdominal adipogenesis.Results:We established a chicken model of abdominal adipocyte differentiation and analyzed miRNA and mRNA expression in abdominal adipocytes at different stages of differentiation(0,12,48,72,and 120 h).A total of 217 differentially expressed miRNAs(DE-miRNAs)and 3520 differentially expressed genes were identified.Target prediction of DE-miRNAs and functional enrichment analysis revealed that the differentially expressed targets were significantly enriched in lipid metabolism-related signaling pathways,including the PPAR signaling and MAPK signaling pathways.A candidate miRNA,gga-miR-106-5p,exhibited decreased expression during the proliferation and differentiation of abdominal preadipocytes and was downregulated in the abdominal adipose tissues of fat chickens compared to that of lean chickens.gga-miR-106-5p was found to inhibit the proliferation and adipogenic differentiation of chicken abdominal preadipocytes.A dual-luciferase reporter assay suggested that the KLF15 gene,which encodes a transcriptional factor,is a direct target of gga-miR-106-5p.gga-miR-106-5p suppressed the posttranscriptional activity of KLF15,which is an activator of abdominal preadipocyte proliferation and differentiation,as determined with gain-and loss-of-function experiments.Conclusions:gga-miR-106-5p functions as an inhibitor of abdominal adipogenesis by targeting the KLF15 gene in chickens.These findings not only improve our understanding of the specific functions of miRNAs in avian adipogenesis but also provide potential targets for the genetic improvement of excessive abdominal fat deposition in poultry.

Effects of Tecar Therapy on Adipose Tissue: Clinical Trial

Introduction: Localised adiposity is one of the leading aesthetic alteration complaints. Tecar therapy has been used in clinical practice for the treatment of fat located in the abdominal region. Objective: The objective is to analyse the effect of Tecar therapy on abdominal adiposity. Methodology: This is a clinical trial in which the participants were 32 female volunteers who have flaccidity and fat located in the abdominal region. The volunteers were divided into three groups, which respectively received applications of the capacitive mode, the resistive mode, and combined therapy of the two modalities. We have performed ten applications and analysed the anthropometric measures, the ultrasonography of the adipose layer, and the level of satisfaction. Results: All groups presented a reduction in perimetry compared to the baseline (p = 0.001). The ultrasonography showed a significant reduction in the combined therapy group compared to the baseline (p = 0.04 and p = 0.03). The level of satisfaction was good in all groups, with few adverse responses. In the bodyweight and percentage of fat analyses, we observed an increase in measurements, probably due to the restriction measures related to the new coronavirus pandemic in 2020. Conclusion: In this present study, we have concluded that Tecar therapy resulted in a reduction of the fat located in the abdominal region.

Effects of One Generation Divergent Selection for Plasma very Low Density Lipoprotein Concentration and Interaction between Genotype and Nutrition on Broiler Chickens

After 1500 male and female broiler chickens from a pure line were selected on the basis of plasma very low density lipoprotein (VLDL) concentration at 8 weeks of age,abdominal fat(AF),liver fat,body weight,meat traits and plasma lipids were determined,and effects of interaction between genotype and nutrition on AF and its correlated traits were studied.1) It was efficient to select against AF on the basis of plasma VLDL concentration.The indirect response of gizzard fat was the greatest,and that of liver fat was the second.AF was decreased,whereas liver fat was increased,feed conversion efficiency during growing period were improved, body weight and meat traits were both not influenced by selecting for low plasma VLDL concentration.2) Nutrition had significant effect on fat deposition and growth performance.Interaction between line and diet had evident effect on AF and its correlated traits.In general,diet had greater effect on H-VLDL line than L-VLDL line,namely chickens of L-VLDL line were less sensitive to diet than those of H-VLDL line.3) Due to high coefficients of variation of plasma VLDL and AF in H-VLDL and L-VLDL lines in the first generation.It will be possible that plasma VLDL is to be selected continuously to breed low fat line of broiler chickens of which the feed conversion efficiency is much better.Moreover,family combined with individual selection should be adopted during future generation selection and breeding because of significant family difference in plasma VLDL concentration.

Follicle-stimulating hormone increases the intramuscular fat content and expression of lipid biosynthesis genes in chicken breast muscle

Intramuscular fat （IMF） is a crucial factor in the quality of chicken meat. The genetic basis underlying it is complex. Follicle-stimulating hormone （FSH）, well-known as an effector in reproductive tissues, was recently discovered to stimulate abdominal fat accumulation in chicken. The effect of FSH on IMF accumulation and the underlying molecular regulatory mechanisms controlling both IMF and abdominal fat deposition in vivo are largely unknown. In this study, two groups of chickens were treated with chicken FSH or a placebo. The lipid content of breast muscle, abdominal fat volume, and serum concentrations of FSH were examined. Related genes implicated in breast muscle and abdominal fat accumulation were also investigated. Compared to the control group, the triglyceride （TG） content of breast muscle and the percentage of abdominal fat in FSH-treated chickens were significantly increased by 64.9% and 56.5% （P〈0.01）, respectively. The FSH content in the serum of FSH-treated chickens was 2.1 times than that of control chickens （P〈0.01）. Results from quantitative real-time polymerase chain reaction （qRT-PCR） assays showed that relative expression levels of fatty acid synthase （FAS）, lipoprotein lipase （LPL）, diacylglycerol acyltransferase 2 （DGAT2）, adipocyte fatty acid binding protein （A-FABP）, and peroxisome proliferator-activated receptor ~ （PPARy） were significantly upregulated in breast muscle following FSH treatment （P〈0.01）. Treatment with FSH also signifi- cantly increased relative expression levels of FAS, LPL, DGA T2, A-FABP, and PPARy in abdominal fat tissue （P〈0.05） The results of principal component analysis （PCA） for gene expression （breast muscle and abdominal fat） showed that the control and FSH treatment groups were well separated, which indicated the reliability of the data. This study demonstrates that FSH plays an important role in IMF accumulation in female chickens, which likely involves the regulation of biosynthesis genes related to lipid metabolism.

Effect of restricted feeding on hen performance,egg quality and organ characteristics of individual laying hens

This study was to assess the impact of permanent or temporary restricted feeding on laying hen production traits,physiology,and egg quality.Two hundred and forty individually housed ISA Brown hens were monitored across 2 phases,assigned to 3 treatments:ad libitum feeding(ALF),temporary restricted feeding(TRF)and permanent restricted feeding(PRF),n=80 hens per treatment.In Phase 1(P1),22 to40 weeks,the TRF and PRF hens were offered 115 g of feed daily.In Phase 2(P2),41 to 46 weeks,the TRF hens were transitioned to ALF status while the ALF and PRF hens remained as in P1.From 35 to 40 weeks,eggs were collected once weekly from 15 hens per treatment and assessed for differences in albumen,yolk,and shell variables.At 45 weeks,10 hens each from the ALF and PRF groups were euthanized and differences in organ characteristics were assessed.In P1,feed intake,feed to egg conversion ratio and body weight(BW)change were lower(P<0.01),while albumen height and Haugh unit were higher(P<0.01)in both PRF and TRF hen treatments compared to hens allocated the ALF treatment.In P2,TRF and ALF hens had a higher egg production and egg mass than PRF(P<0.01)than ALF.Body weight change in P2 was higher in TRF and similar in both ALF and PRF,while feed intake and feed conversion ratio were higher in TRF followed by ALF and least in the PRF treatment group(P<0.01).At 45 weeks ALF hens had a greater abdominal fat pad weight and fatty liver haemorrhagic syndrome lesion score compared to PRF.Restricting hens to 115 g of feed per day from point of lay restrained BW,improved feed conversion ratio and albumen quality and reduced abdominal fat pad deposition and clinical signs of fatty liver haemorrhagic syndrome in individually housed laying hens.