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Conjugated linoleic acids (CLA) are known to reduce body fat and plasma lipids in animal models. This study examined the short-term effects of 2 biologically active isomers of CLA (cis-9, trans-11 and trans-10, cis-12) on lipid-metabolizing genes and high-density lipoprotein (HDL) cholesterol production in cultured HepG2 cells. Steady-state levels of acyl CoA oxidase (ACO); 3-hydroxy, 3-methylglutaryl CoA reductase (HMG-R); and apolipoprotein A-I (Apo A-I) mRNA were examined after 24-hour incubation in the absence (control) or presence of 100 microm each of linoleic acid (LA), cis-9, trans-11 CLA or trans-10, cis-12 CLA. Concentrations of ACO and HMG-R mRNA transcripts were increased in HepG2 cells treated with trans-10, cis-12 CLA. Incubation with MK886, a specific PPAR[alpha] inhibitor, had minimal effect on basal or CLA-induced gene expression in HepG2 cells. The cis-9, trans-11, but not trans-10, cis-12, CLA decreased HDL cholesterol concentration in cell-conditioned media. There was no apparent relationship between Apo A-I and HDL cholesterol responses to fatty acids. Results indicate that CLA may control peroxisomal oxidation of fatty acids through up-regulation of hepatic ACO gene expression. The physiological relevance of CLA effect on HMG-R mRNA content in the liver is yet to be determined. [ABSTRACT FROM AUTHOR]

Research Highlights: Wire ramp model reproducibly induced lameness/BCO in broilers.Treatments did not affect growth, but phytase with stimbiotic significantly reduced BCO.Phytase increased circulating inositol, and wire flooring decreased bone inositol.

Divergent selection of broilers for water conversion ratio has established and high-(HWE) and low- water efficient (LWE) broiler lines. Two 2 × 2 factorial experiments were conducted to assess the gene expression profile of systems involved in renal water homeostasis. In Exp. 1, male and female HWE and LWE broilers were individually phenotyped between 4 and 6 wks of age to determine growth performance and water conversion ratio (g water intake/g body weight gain). Kidney samples were obtained from 5 males and 5 females from each line. In Exp. 2, HWE and modern random bred (MRB) broilers were placed in 12 controlled-environmental chambers (2 floor pens/chamber, 6 chambers/line, 11 birds per pen, 132 birds/line) on day of hatch. The broilers were brooded at thermoneutral temperatures from 0 to 4 wks. From 4 to 7 wks, broilers were maintained at thermoneutral (TN, 25 °C) or exposed to cyclic heat stress (HS, 35 °C, 8h/day) conditions. Body weight, feed intake, and water intake were recorded. Kidney samples were collected, flash frozen in liquid nitrogen, and kept at -80 °C for gene expression analysis. Data were analyzed by Two-way ANOVA and means compared by Tukey's HSD multiple comparison test. Molecular analyses from Exp. 1 showed that the renal expression of arginine vasopressin (AVP), angiotensinogen (AGT), angiotensin II receptor type 1 and 2 (AT1/2), sodium-potassium ATPase subunit B1 (ATP1B1), and aquaporin 3 (AQP3) were upregulated in HWE compared to the LWE line. In contrast, mRNA expression of mesotocin receptor (MTR), AT1/2, AQP1/2, and occludin were significantly higher in females than in males. In Exp. 2, target genes were regulated in environment and/or line-dependent manner. The renal expression of heat shock proteins 70 and 90, AVP receptor 2 (AVPR2), AGT, renin, AT1/2, and AQP1was significantly upregulated in HS compared to TN birds, however AVPR2 expression was significantly higher in HWE compared to MRB birds. Together, the up-regulation of AVP, the renin-angiotensin system (RAS), and AQP in HWE, female, or under HS conditions suggests a better renal water reabsorption to support water use efficiency., Competing Interests: Declaration of competing interest Authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024. Published by Elsevier Inc.)

In the broiler industry, intensive genetic selection has been placed on muscle growth which has undesirably led to increased fat accretion. Models of chicken preadipocyte differentiation in vitro have conventionally used incubators without the ability to control oxygen (O 2 ) tension; thus, the cells are exposed to atmospheric (∼20-21%) O 2 , which is supraphysiological compared to the O 2 tension within adipose tissue. The objective of this study was to investigate embryonic broiler preadipocyte differentiation at different O 2 tensions, including atmospheric (20%), physiological (5%), and hypoxic (1%). Culture at 1% O 2 resulted in increased abundance of HIF1α, a canonical protein stabilized during hypoxia, thus confirming effectiveness of the treatment. Increased accumulation of lipid was observed in preadipocytes cultured in adipogenic differentiation medium compared to the control medium. When considering oxygen tension, lipid accumulation was increased in preadipocytes that were cultured in differentiation medium at 20% O 2 compared to 5% or 1% O 2 . Furthermore, abundance of transcripts related to fatty acid transport and adipogenesis, fatty acid binding protein 4 (FABP4) and peroxisome proliferator-activated receptor gamma (PPARγ), were increased in differentiated preadipocytes cultured at 20% O 2 compared to 5% or 1% O 2 . Abundance of transcripts related to lipid synthesis and oxidation, acyl-CoA synthetase long chain family member 1 (ACSL1) and carnitine palmitoyltransferase 1A (CPT1A), were increased in the differentiation cultures compared to the control cultures. Abundance of glutathione peroxidase 4 (GPX4) was increased in all the differentiation cultures compared to the controls, regardless of oxygen tension; however, differences in the abundance of other antioxidant enzymes were not observed. Overall, exposure to atmospheric oxygen tension promotes lipid accumulation within chicken preadipocytes, which may need to be considered when developing in vitro models of this process., Competing Interests: Declaration of competing interest The authors declare that no conflict of interest exists., (Copyright © 2024. Published by Elsevier Inc.)

Background: Water scarcity is a current, significant global concern that will only increase under the pressure of climate change. Improving water efficiency of poultry is a new and promising area to help temper agriculture's future impact on fresh water availability. Here, we explored the effects of acute heat stress (HS) on circulating stress and inflammatory markers in 2 lines of broilers divergently selected for water efficiency., Methods: Male chicks from low (LWE) and high water efficient (HWE) lines were raised in 12 environmental chambers (2 pens/chamber, 6 chambers/line, 20 birds/pen) under normal conditions until day 28. On day 29, birds were subjected to thermoneutral (TN, 25 °C) or HS (36 °C) conditions, resulting in four treatments (2 lines × 2 environmental conditions). After 3 h of HS, whole blood was collected (8 birds per line × environment) and analyzed for target gene expression and plasma cytokine levels. Data were analyzed by 2-way ANOVA, with line, environment, and their interaction as main factors, and means were compared using Tukey's multiple range test., Results: Gene expression of heat shock protein (HSP) 27, HSP70, interleukin (IL)-6, IL-18, c-reactive protein (CRP), tumor necrosis factor-α (TNFα), C-C motif chemokine ligand 4 (CCL4), CCL20, nucleotide-binding domain, leucine-rich repeat (NLR) family pyrin domain containing 3 (NLRP3), NLR family CARD domain containing 5 (NLRC5), and NLR family member X1 (NLRX1) were increased by HS, with no differences between the lines. HSP70, IL-10, and NLRC3 were lower in the HWE as compared to the LWE lines. Additionally, there were interactive effects between line and environment for HSP90, IL-4, and CCL4, where HS induced HSP90 expression in the LWE only, and IL-4 and CCL4 in HWE only. Arginine vasopressin (AVP) gene expression was significantly lower in the whole blood of the HWE line; however, plasma protein levels were not different., Conclusions: Overall, most of the effects seen on cyto (chemokines) and inflammatory markers were due to acute HS, with only a few genes differentially regulated between the lines. This likely indicates that the divergent selection for water efficiency for four generations did not elicit changes in inflammation and stress molecular signatures., (© 2024 The Author(s). Published by IMR Press.)

Heat stress (HS) is a global serious issue in the poultry industry with numerous adverse effects, including increased stress, depressed feed intake (FI), poor growth performance and higher mortality. Herbal adaptogens, plant extracts considered as stress response modifiers, are metabolic regulators that improve an organism's ability to adapt to and minimize damage from environmental stresses. Previously, we showed that herbal adaptogen supplementation increased FI and body weight (BW) of broiler (meat-type) chickens reared under HS conditions. Therefore, we hypothesized that these effects may be mediated through modulation of hypothalamic feeding-related neuropeptides. Male Cobb 500 chicks were reared in 12 environmental chambers with three diets: a corn-soybean-based diet (C) and two herbal adaptogen-supplemented diets at 500 g/1000 kg (NR-PHY-500) and 1 kg/1000 kg (NR-PHY-1000). Broilers in 9 chambers were exposed to chronic cyclic HS (35 °C for 8 h/day) from d29 to d42, while 3 chambers were maintained at 24 °C (thermoneutral, TN) for all 42 days. Hypothalamic samples were collected on d42 from each group, both before the onset of HS (Pre-HS) that day and after 3 h of HS (post-HS). Hypothalamic expressions of neuropeptide Y (NPY) receptors Y4 and Y7, Corticotropin-releasing hormone (CRH), orexin receptor 1 (ORXR1), melanocortin receptors (MC1R, MC4R, and MC5R), visfatin and neurosecretory protein GL (NPGL) genes were significantly upregulated by adaptogen supplementation. The hypothalamic expression of MC2R was affect by period, with a significant upregulation during post-HS phase. There was a significant period by treatment interaction for hypothalamic orexin and adiponectin expression. The hypothalamic expression of NPY, Y1, Y2, Y5, Y6, proopiomelanocortin (POMC), cocaine and amphetamine regulated transcript (CART), agouti-related peptide (AgRP), ORXR2, AdipR1/2, MC3R, and ghrelin was not affected by diet supplementation nor by HS exposure. In conclusion, these findings suggest that in-feed supplementation of adaptogen might improve FI and growth via modulation of hypothalamic feeding-related neuropeptides in heat-stressed broilers., (Copyright © 2023. Published by Elsevier Ltd.)

A total of 720 male Cobb 500 broiler chicks were used in a 5 treatment and 8 replicate experiment to explore dynamic changes in blood metabolites in response to short-term nutrient depletion and repletion. Day old chicks were offered a corn and soybean meal-based common starter diet from d1 to 14 that was formulated to meet all nutrient requirements of the birds. From d15 to 17, the experimental diets were offered, before returning all groups to a common diet from d18 to 20, at which point the experiment was terminated. A total of 5 experimental diets were designed. A standard grower diet served as a control and was offered to 1 of the 5 groups of chicks. The additional 4 experimental groups comprised diets that were low in digestible phosphorus (P), total calcium (Ca), crude protein and digestible amino acids (AA) or apparent metabolizable energy (AME). The common grower diet that was offered from d18-20 was designed to be nutritionally complete and was intended to explore dynamic response to nutrient repletion. Blood was drawn from 8 chicks per treatment at time 0 (immediately prior to introduction of the experimental diets) and then again 3, 6, 12, 24, and 48h after introduction of the nutrient depleted diets. Additionally, blood was drawn 3, 6, 12, 24, and 48h after the introduction of the nutritionally complete common grower diet. Chicks were not sampled more than once. Feed intake, body weight and feed conversion ratio (FCR) were assessed on d14, 17, and 20. Blood metabolites were analyzed using the iSTAT Alinity V handheld blood analyzer, the Vetscan VS2 Chemistry Analyzer and the iCheck Carotene Photometer. Live performance metrics were not affected by the short-term nutrient depletion and all chicks grew normally throughout the experiment. The diet with low digestible P generated a rapid temporary decrease in plasma P and an increase in plasma Ca, that were returned to baseline following the re-introduction of the common grower feed. Introduction of the diet with low total Ca resulted in a significant increase in plasma P, effects which were also mitigated during the nutrient repletion phase. Total plasma protein, albumin and uric acid (UA) were decreased, and plasma glucose increased, in the chicks that received the diet with low crude protein and digestible AA. There was a delayed increase in aspartate amino transaminase (AST) associated with the diets with low digestible P and low AME. These results demonstrate the capacity of blood biochemistry to adapt to quantitative and qualitative changes in nutrient intake. Point-of-care analysis of blood biomarkers offers nutritionists a valuable opportunity to calibrate nutritional matrices for common dietary ingredients, zootechnical feed additives and to optimize diet phase changes. It can be concluded that many blood biomarkers are plastic to changes in diet nutrient density and offer an objective index for optimization of nutritional programs for commercial broiler production., Competing Interests: DISCLOSURES A. J. Cowieson, C. A. Phillips, and E. Papadopoulou are employed by DSM Animal Nutrition. DSM Animal Nutrition had no role in conducting the research or in generating the data. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Leg health is a significant economic and welfare concern for the poultry industry. Current methods of detection rely on visual assessment of the legs and gait scores and bone scoring during necropsy for full characterization. Additionally, the current scoring of femurs only examines the external surface of the femoral head. Through the use of the dual-energy X-ray absorptiometry (DXA) imaging system, we show the presence of a necrotic region in the femurs that would otherwise be considered healthy based on the current evaluation procedures. Importantly, these lesions were present in almost 60% (22 of 37) of femurs that scored normal for femoral head necrosis (FHN). Additionally, these femurs showed greater bone mineral content (BMC) relative to weight compared to their counterparts with no lucent lesions (6.95% ± 0.20% vs. 6.26% ± 0.25; p = 0.038). Identification of these lesions presents both a challenge and an opportunity. These subclinical lesions are likely to be missed in routine scoring procedures for FHN and can inadvertently impact the characterization of the disease and genetic selection programs. Furthermore, this imaging system can be used for in vivo , ex vivo , and embryonic (egg) studies and, therefore, constitutes a potential non-invasive method for early detection of bone lesions in chickens and other avian species., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Ramser, Greene, Wideman and Dridi.)

Genetic selection for high growth rate has resulted in spectacular progress in feed efficiency in chickens. As feed intake and water consumption (WC) are associated and both are affected by environmental conditions, we evaluated WC and its hypothalamic regulation in three broiler-based research lines and their ancestor jungle fowl (JF) under heat stress (HS) conditions. Slow growing ACRB, moderate growing 95RB, fast growing MRB, and JF were exposed to daily chronic cyclic HS (36 °C, 9 h/d) or thermoneutral temperature (24 °C). HS increased WC in the MRB only. Arginine vasopressin (AVP) mRNA levels were decreased by HS in the MRB. Within the renin-angiotensin-aldosterone system (RAAS) system, renin expression was increased by HS in the JF, ACRB, and 95RB, while angiotensin I-converting enzyme (ACE), angiotensin II receptors (type 1, AT1, and type 2, AT2) were affected by line. The expression of aquaporin (AQP2, 7, 9, 10, 11, and 12) genes was upregulated by HS, whereas AQP4 and AQP5 expressions were influenced by line. miRNA processing components (Dicer1, Ago2, Drosha) were significantly different among the lines, but were unaffected by HS. In summary, this is the first report showing the effect of HS on hypothalamic water channel- and noncoding RNA biogenesis-related genes in modern chicken populations and their ancestor JF. These results provide a novel framework for future research to identify new molecular mechanisms and signatures involved in water homeostasis and adaptation to HS., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier B.V.)

Freshwater ecosystems are biologically important habitats that provide many ecosystem services. Calcium concentration and pH are two key variables that are linked to multiple chemical processes in these environments, influence the biology of organisms from diverse taxa, and can be important factors affecting the distribution of native and non-native species. However, it can be challenging to obtain high-resolution data for these variables at regional and national scales. To address this data gap, water quality data for lakes and rivers in Canada and the continental USA were compiled and used to generate high-resolution (10 × 10 km) interpolated raster layers, after comparing multiple spatial interpolation approaches. This is the first time that such data have been made available at this scale and resolution, providing a valuable resource for research, including projects evaluating risks from environmental change, pollution, and invasive species. This will aid the development of conservation and management strategies for these vital habitats., (© 2024. Crown.)

Powered by consumer taste, value, and preferences, natural products including phytogenics and algae are increasingly and separately used in the food systems where they have been reported to improve growth performance in poultry and livestock. The present study aimed to determine the effects of a new feed additive, microencapsulated NUQO© NEX, which contains a combination of phytogenic and phycogenic, on broiler growth performance, blood chemistry, bone health, meat quality and sensory profile. Male Cobb500 chicks ( n  = 1,197) were fed a 3-phase feeding intervals; 1-14d starter, 15-28d grower, and 29-40d finisher. The dietary treatments included a corn-soy basal Control (CON), basal diet supplemented with NUQO© NEX at 100 g/ton from 1 to 28d then 75 g/ton from d 28 to 40 (NEX75), and basal diet supplemented with NUQO© NEX at 100 g/ton from 1 to 40d (NEX100). The NEX100 supplemented birds had 62 g more BWG increase and 2.1-point improvement in FCR compared with CON in the finisher and overall growth phase ( p  < 0.05), respectively. Day 40 processing body weights and carcass weights were heavier for the NEX100 supplemented birds ( p  < 0.05). The incidences of muscle myopathies were also higher in NEX treatments, which could be associated with the heavier weights, but the differences were not detected to be significant. The NEX75 breast filets had more yellowness than other dietary treatments ( p  = 0.003) and the NEX 100 treatment reduced the levels of breast filet TBARS at 7 days-post harvest ( p  = 0.053). Finally, both NEX treatments reduced the incidence of severe bone (tibia and femur) lesions. In conclusion, the supplementation of the phytogenic NUQO© NEX improved finisher performance parameters, whole phase FCR, processing carcass weights, and breast filet yellowness, at varying inclusion levels., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Mullenix, Greene, Ramser, Maynard and Dridi.)

The objective of the present study was to determine the effect of a novel (4th generation) phytase supplementation as well as its mode of action on growth, meat quality, and incidence of muscle myopathies. One-day old male broilers ( n = 720) were weighed and randomly allocated to 30 floor pens (24 birds/pen) with 10 replicate pens per treatment. Three diets were fed from hatch to 56- days-old: a 3-phase corn-soy based diet as a positive control (PC); a negative control (NC) formulated to be isocaloric and isonitrogenous to the PC and with a reduction in Ca and available P, respectively; and the NC supplemented with 2,000 phytase units per kg of diet (NC + P). At the conclusion of the experiment, birds fed with NC + P diet were significantly heavier and had 2.1- and 4.2-points better feed conversion ratio (FCR) compared to birds offered NC and PC diets, respectively. Processing data showed that phytase supplementation increased live weight, hot carcass without giblets, wings, tender, and skin-on drum and thigh compared to both NC and PC diets. Macroscopic scoring showed that birds fed the NC + P diet had lower woody breast (WB) severity compared to those fed the PC and NC diets, however there was no effect on white striping (WS) incidence and meat quality parameters (pH, drip loss, meat color). To delineate its mode of action, iSTAT showed that blood glucose concentrations were significantly lower in birds fed NC + P diet compared to those offered PC and NC diets, suggesting a better glucose uptake. In support, molecular analyses demonstrated that the breast muscle expression (mRNA and protein) of glucose transporter 1 (GLUT1) and glucokinase (GK) was significantly upregulated in birds fed NC + P diet compared to those fed the NC and PC diets. The expression of mitochondrial ATP synthase F0 subunit 8 (MT-ATP8) was significantly upregulated in NC + P compared to other groups, indicating intracellular ATP abundance for anabolic pathways. This was confirmed by the reduced level of phosphorylated-AMP-activated protein kinase (AMPKα1/2) at Thr172 site, upregulation of glycogen synthase (GYS1) gene and activation of mechanistic target of rapamycin and ribosomal protein S6 kinase (mTOR-P70S6K) pathway. In conclusion, this is the first report showing that in-feed supplementation of the novel phytase improves growth performance and reduces WB severity in broilers potentially through enhancement of glucose uptake, glycolysis, and intracellular ATP production, which used for muscle glycogenesis and protein synthesis., Competing Interests: Authors Carrie L. Walk and Nelson Ward were employed by DSM Nutritional Products. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Walk, Mullenix, Maynard, Greene, Maynard, Ward and Dridi.)

With climate change, selection for water efficiency and heat resilience are vitally important. We undertook this study to determine the effect of chronic cyclic heat stress (HS) on the hypothalamic expression profile of water homeostasis-associated markers in high (HWE)- and low (LWE)-water efficient chicken lines. HS significantly elevated core body temperatures of both lines. However, the amplitude was higher by 0.5-1°C in HWE compared to their LWE counterparts. HWE line drank significantly less water than LWE during both thermoneutral (TN) and HS conditions, and HS increased water intake in both lines with pronounced magnitude in LWE birds. HWE had better feed conversion ratio (FCR), water conversion ratio (WCR), and water to feed intake ratio. At the molecular level, the overall hypothalamic expression of aquaporins (AQP8 and AQP12), arginine vasopressin (AVP) and its related receptor AVP2R, angiotensinogen (AGT), angiotensin II receptor type 1 (AT1), and calbindin 2 (CALB2) were significantly lower; however, CALB1 mRNA and AQP2 protein levels were higher in HWE compared to LWE line. Compared to TN conditions, HS exposure significantly increased mRNA abundances of AQPs (8, 12), AVPR1a, natriuretic peptide A (NPPA), angiotensin I-converting enzyme (ACE), CALB1 and 2, and transient receptor potential cation channel subfamily V member 1 and 4 (TRPV1 and TRPV4) as well as the protein levels of AQP2, however it decreased that of AQP4 gene expression. A significant line by environment interaction was observed in several hypothalamic genes. Heat stress significantly upregulated AQP2 and SCT at mRNA levels and AQP1 and AQP3 at both mRNA and protein levels, but it downregulated that of AQP4 protein only in LWE birds. In HWE broilers, however, HS upregulated the hypothalamic expression of renin (REN) and AVPR1b genes and AQP5 proteins, but it downregulated that of AQP3 protein. The hypothalamic expression of AQP (5, 7, 10, and 11) genes was increased by HS in both chicken lines. In summary, this is the first report showing improvement of growth performances in HWE birds. The hypothalamic expression of several genes was affected in a line- and/or environment-dependent manner, revealing potential molecular signatures for water efficiency and/or heat tolerance in chickens., (© 2024 The Authors. Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society.)

Heat stress (HS) is one of the most challenging stressors to poultry production sustainability. The adverse effects of HS range from feed intake and growth depression to alteration of meat quality and safety. As phytase supplementation is known to improve nutrient utilization and consequently growth, we undertook the present study to evaluate the effects of dietary phytase on growth and meat quality in heat-stressed broilers. A total of 720 day-old hatch Cobb 500 chicks were assigned to 24 pens within controlled environmental chambers and fed three diets: Negative Control (NC), Positive Control (PC), and NC diet supplemented with 2000 phytase units (FTU)/kg) of quantum blue (QB). On day 29, birds were exposed to two environmental conditions: thermoneutral (TN, 25 °C) or cyclic heat stress (HS, 35 °C, 8 h/d from 9 a.m. to 5 p.m.) in a 3 × 2 factorial design. Feed intake (FI), water consumption (WI), body weight (BW), and mortality were recorded. On day 42, birds were processed, carcass parts were weighed, and meat quality was assessed. Breast tissues were collected for determining the expression of target genes by real-time quantitative PCR using the 2 -ΔΔCt method. HS significantly increased core body temperature, reduced feed intake and BW, increased water intake (WI), elevated blood parameters (pH, SO 2 , and iCa), and decreased blood pCO 2 . HS reduced the incidence of woody breast (WB) and white striping (WS), significantly decreased drip loss, and increased both 4- and 24-h postmortem pH. Instrumental L* and b* values were reduced ( p < 0.05) by the environmental temperature at both 4- and 24-h postmortem. QB supplementation reduced birds' core body temperature induced by HS and improved the FCR and water conversion ratio (WCR) by 1- and 0.5-point, respectively, compared to PC under HS. QB increased blood SO 2 and reduced the severity of WB and WS under TN conditions, but it increased it under an HS environment. The abovementioned effects were probably mediated through the modulation of monocarboxylate transporter 1, heat shock protein 70, mitogen-activated protein kinase, and/or glutathione peroxidase 1 gene expression, however, further mechanistic studies are warranted. In summary, QB supplementation improved growth performance and reduced muscle myopathy incidence under TN conditions. Under HS conditions, however, QB improved growth performance but increased the incidence of muscle myopathies. Therefore, further QB titration studies are needed.

Essential oils (EO) affect performance, intestinal integrity, bone mineralization, and meat quality in broiler chickens subjected to cyclic heat stress (HS). Day-of-hatch Cobb 500 male broiler chicks ( n = 475) were randomly divided into four groups. Group 1: No heat stress (Thermoneutral) + control diets with no antibiotics; Group 2: heat stress control + control diets; Group 3: heat stress + control diets supplemented with thymol chemotype (45 ppm) and herbal betaine (150 ppm) formulation EO1; Group 4: heat stress + control diets supplemented with phellandrene (45 ppm) and herbal betaine (150 ppm) formulation EO2. From day 10-42, the heat stress groups were exposed to cyclic HS at 35°C for 12 h (8:00-20:00). BW, BWG, FI, and FCRc were measured at d 0, 10, 28, and 42. Chickens were orally gavaged with FITC-d on days 10 (before heat stress) and 42. Morphometric analysis of duodenum and ileum samples and bone mineralization of tibias were done. Meat quality was assessed on day 43 with ten chickens per pen per treatment. Heat stress reduced BW by day 28 ( p < 0.05) compared to thermoneutral chickens. At the end of the trial, chickens that received both formulations of EO1 and EO2 had significantly higher BW than HS control chickens. A similar trend was observed for BWG. FCRc was impaired by EO2 supplementation. There was a significant increase in total mortality in EO2 compared with EO1 EO1 chickens had lower FITC-d concentrations at day 42 than the HS control. In addition, EO1 treatment is not statistically different if compared to EO2 and thermoneutral. Control HS broilers had significantly lower tibia breaking strength and total ash at day 42 than heat-stressed chickens supplemented with EO1 and EO2. Heat stress affected intestinal morphology more than thermoneutral chickens. EO1 and EO2 improved intestinal morphology in heat-stressed chickens. Woody breast and white striping were more common in thermoneutral chickens than heat stress chickens. In conclusion, the EO-containing diet could improve broiler chicken growth during cyclic heat stress, becoming increasingly relevant in antibiotic-free production in harsh climates., Competing Interests: Authors AU, BM, and JA-I, were employed by the company Promitec. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Señas-Cuesta, Stein, Latorre, Maynard, Hernandez-Velasco, Petrone-Garcia, Greene, Coles, Gray, Laverty, Martin, Loeza, Uribe, Martínez, Angel-Isaza, Graham, Owens, Hargis and Tellez-Isaias.)

Background: The limitations to prolonged spaceflight include unloading-induced atrophy of the musculoskeletal system which may be enhanced by exposure to the space radiation environment. Previous results have concluded that partial gravity, comparable to the Lunar surface, may have detrimental effects on skeletal muscle. However, little is known if these outcomes are exacerbated by exposure to low-dose rate, high-energy radiation common to the space environment. Therefore, the present study sought to determine the impact of highly charge, high-energy (HZE) radiation on skeletal muscle when combined with partial weightbearing to simulate Lunar gravity. We hypothesized that partial unloading would compromise skeletal muscle and these effects would be exacerbated by radiation exposure., Methods: For month old female BALB/cByJ mice were -assigned to one of 2 groups; either full weight bearing (Cage Controls, CC) or partial weight bearing equal to 1/6th bodyweight (G/6). Both groups were then divided to receive either a single whole body absorbed dose of 0.5 Gy of 300 MeV 28 Si ions (RAD) or a sham treatment (SHAM). Radiation exposure experiments were performed at the NASA Space Radiation Laboratory (NSRL) located at Brookhaven National Laboratory on Day 0, followed by 21 d of CC or G/6 loading. Muscles of the hind limb were used to measure protein synthesis and other histological measures., Results: Twenty-one days of Lunar gravity (G/6) resulted in lower soleus, plantaris, and gastrocnemius muscle mass. Radiation exposure did not further impact muscle mass. 28 Si exposure in normal ambulatory animals (RAD+CC) did not impact gastrocnemius muscle mass when compared to SHAM+CC (p>0.05), but did affect the soleus, where mass was higher following radiation compared to SHAM (p<0.05). Mixed gastrocnemius muscle protein synthesis was lower in both unloading groups. Fiber type composition transitioned towards a faster isoform with partial unloading and was not further impacted by radiation. The combined effects of partial loading and radiation partially mitigated fiber cross-sectional area when compared to partial loading alone. Radiation and G/6 reduced the total number of myonuclei per fiber while leading to elevated BrdU content of skeletal muscle. Similarly, unloading and radiation resulted in higher collagen content of muscle when compared to controls, but the effects of combined exposure were not additive., Conclusions: The results of this study confirm that partial weightbearing causes muscle atrophy, in part due to reductions of muscle protein synthesis in the soleus and gastrocnemius as well as reduced peripheral nuclei per fiber. Additionally, we present novel data illustrating 28 Si exposure reduced nuclei in muscle fibers despite higher satellite cell fusion, but did not exacerbate muscle atrophy, CSA changes, or collagen content. In conclusion, both partial loading and HZE radiation can negatively impact muscle morphology., Competing Interests: Declaration of Competing Interest Y.L. is currently working at GlaxoSmithKline. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 The Committee on Space Research (COSPAR). Published by Elsevier B.V. All rights reserved.)

Bacterial Chondronecrosis with Osteomyelitis (BCO) is a specific cause of lameness in commercial fast-growing broiler (meat-type) chickens and represents significant economic, health, and wellbeing burdens. However, the molecular mechanisms underlying the pathogenesis remain poorly understood. This study represents the first comprehensive characterization of the proximal tibia proteome from healthy and BCO chickens. Among a total of 547 proteins identified, 222 were differentially expressed (DE) with 158 up- and 64 down-regulated proteins in tibia of BCO vs. normal chickens. Biological function analysis using Ingenuity Pathways showed that the DE proteins were associated with a variety of diseases including cell death, organismal injury, skeletal and muscular disorder, immunological and inflammatory diseases. Canonical pathway and protein-protein interaction network analysis indicated that these DE proteins were involved in stress response, unfolded protein response, ribosomal protein dysfunction, and actin cytoskeleton signaling. Further, we identified proteins involved in bone resorption (osteoclast-stimulating factor 1, OSFT1) and bone structural integrity (collagen alpha-2 (I) chain, COL2A1), as potential key proteins involved in bone attrition. These results provide new insights by identifying key protein candidates involved in BCO and will have significant impact in understanding BCO pathogenesis., (© 2023. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Although broiler (meat-type) chickens are one of the most efficient protein sources that supports the livelihoods and food security of billions of people worldwide, they are facing several challenges. Due to its unknown etiology and heavy economic impact, woody breast (WB) myopathy is one of the most challenging problems facing the poultry industry, and for which there is no effective solution. Here, using a primary chicken myotube culture model, we show that hypoxia and endoplasmic reticulum (ER) stress are an integral component of the etiology of the myopathy. Multiple components of the ER stress response are significantly upregulated in WB as compared with normal muscle, and this response was mimicked by hypoxic conditions in chicken primary myotube culture. In addition, apoptotic pathways were activated as indicated by increases in active caspase 3 protein levels in both WB-affected tissues and hypoxic myotube culture, and caspase 3 activity and apoptosis in hypoxic myotube culture. Finally, as a phenotypic hallmark of WB is enhanced fibrosis and increased collagen aggregation, here, we show that hypoxic conditions increase collagen 1A1 and 1A2 gene expression, as well as collagen 1 protein levels in primary myotubes. These effects were partially reversed by tauroursodeoxycholic acid (TUDCA), an ER-stress inhibitor, in myotube culture. Taken together, these findings indicate that hypoxia and ER stress are present in WB, hypoxia can upregulate the cell death arm of the unfolded protein response (UPR) and lead to collagen production in a culture model of WB. This opens new vistas for potential mechanistic targets for future effective interventions to mitigate this myopathy.

This experiment compared the effects of 2 chronic heat stress (HS) models, constant (coHS), and cyclic (cyHS), on broiler performance, carcass characteristics, and meat quality. A total of 720 male chicks from a Cobb 500 line were placed in 12 environmentally controlled chambers divided into 2 pens of 30 birds. Before the experimental HS models were applied, chamber temperatures were gradually decreased from 32°C at placement to 24°C on d 20. From 20 to 41 d, 4 chambers were set to 35°C (coHS), and 4 chambers were set to 35°C for 12 h and 24°C for the next 12 h (cyHS). Four thermoneutral chambers were maintained at 24°C with half of the birds pair-fed to equalize feed intake (FI) with coHS birds (TN-coPF) and half fed ad-libitum (TN-al). From 20 to 41 d, FI and BW gain (BWG) of cyHS, coHS and TN-coPF birds were decreased (P < 0.001), whereas feed conversion ratio (FCR) was increased (P < 0.001) for coHS and TN-coPF birds compared with TN-al birds. The overall BWG and FCR of coHS birds were lower (P < 0.001) than TN-coPF birds. Both HS models reduced (P < 0.001) carcass weight, pectoralis major yield, total breast meat yield, and increased (P < 0.001) wing yield relative to TN-al birds, with each of these measurements more impacted by coHS than by cyHS. Pair-fed birds had lower (P < 0.001) fat pad and a higher total breast meat yield than coHS birds. They also had the lowest (P < 0.001) pectoralis major ultimate pH and yellowness, and these parameters were lower (P < 0.001) for coHS birds than for TN-al birds. Both HS models reduced (P < 0.001) the incidence of woody breast and white striping. Thus, these data indicate that the detrimental effects of HS cannot be entirely explained by reduced FI and that HS per se affects metabolic pathways associated with muscle and lipid accretion in broilers., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Commercial broilers have been selected for high growth rate and productivity; however, this has negatively impacted their susceptibility to heat stress (HS). Insight into the molecular mechanisms underlying this vulnerability can help design targeted strategies for improvement of HS tolerance. Red blood cells (RBC) and white blood cells (WBC) were isolated from red jungle fowl and 4 lines of commercial modern broilers. Lines A and B are considered standard-yielding lines, whereas Lines C and D are high-yielding. Cells were cultured at either 37°C or 45°C for 2 h to induce heat stress (HS). Gene expression of cytokines, chemokines, and inflammasome components were measured. Heat shock proteins 27 and 70 (HSPs) in RBC were significantly affected by line (P < 0.05), whereas HSP27 and 60 were affected by temperature (P < 0.05). In WBC, there was a significant line effect on HSP gene expression (P < 0.05), and a significant increase (P < 0.05) in HSP90 in Line D in HS compared to TN conditions. In RBC, there was a main effect of HS on TNFα, CCL4, and CCLL4 (P < 0.05). HS significantly increased IL-8L1 (>30-fold, P < 0.0001) in Line C. Inflammasome genes (NLRP3, NLRC5 and NLRC3) were significantly affected by the line studied (P < 0.05). In WBC, the effect of line was significant for all cytokines, chemokines, and inflammasome components studied (P < 0.05). To examine the mechanical properties of isolated RBC from the 4 commercial lines and jungle fowl, RBC were placed into nematic liquid crystals, where Lines B and D were the most strained, and Line A and the jungle fowl were the least strained. Together, these findings indicate not only the dynamic nature of circulating cells, but the differences in the stress and inflammatory response among commercially available lines and their common ancestor. These profiles have the potential to serve as a future marker for stress responses in broilers, though further study is warranted., (Published by Elsevier Inc.)

Heat stress (HS) is devastating to poultry production sustainability worldwide. In addition to its adverse effects on growth, welfare, meat quality, and mortality, HS alters the gut integrity, leading to dysbiosis and leaky gut syndrome; however, the underlying mechanisms are not fully defined. Here, we used a high-throughput mass spectrometric metabolomics approach to probe the metabolite profile in the duodenum of modern broilers exposed to acute (AHS, 2 h) or chronic cyclic (CHS, 8 h/day for 2 weeks) HS in comparison with thermoneutral (TN) and pair-fed birds. Ultra high performance liquid chromatography coupled with high resolution mass spectrometry (UHPLC-HRMS) identified a total of 178 known metabolites. The trajectory analysis of the principal component analysis (PCA) score plots (both 2D and 3D maps) showed clear separation between TN and each treated group, indicating a unique duodenal metabolite profile in HS birds. Within the HS groups, partial least squares discriminant analysis (PLS-DA) displayed different clusters when comparing metabolite profiles from AHS and CHS birds, suggesting that the metabolite signatures were also dependent on HS duration. To gain biologically related molecule networks, the above identified duodenal metabolites were mapped into the Ingenuity Pathway Analysis (IPA) knowledge-base and analyzed to outline the most enriched biological functions. Several common and specific top canonical pathways were generated. Specifically, the adenosine nucleotide degradation and dopamine degradation pathways were specific for the AHS group; however, the UDP-D-xylose and UDP-D-glucuronate biosynthesis pathways were generated only for the CHS group. The top diseases enriched by the IPA core analysis for the DA metabolites, including cancer, organismal (GI) injury, hematological, cardiovascular, developmental, hereditary, and neurological disorders, were group-specific. The top altered molecular and cellular functions were amino acid metabolism, molecular transport, small molecule biochemistry, protein synthesis, cell death and survival, and DNA damage and repair. The IPA-causal network predicted that the upstream regulators (carnitine palmitoyltransferase 1B, CPT1B; histone deacetylase 11, HDAC11; carbonic anhydrase 9, CA9; interleukin 37, IL37; glycine N-methyl transferase, GNMT; GATA4) and the downstream mediators (mitogen-activated protein kinases, MAPKs; superoxide dismutase, SOD) were altered in the HS groups. Taken together, these data showed that, independently of feed intake depression, HS induced significant changes in the duodenal metabolite profile in a duration-dependent manner and identified a potential duodenal signature for HS.

The goal of this research was to assess cyclic heat stress on gut permeability, bone mineralization, and meat quality in chickens. Two separate trials were directed. 320 day-of-hatch Cobb 500 male chicks were randomly assigned to four thermoneutral (TN) and four cyclic heat stress (HS) chambers with two pens each, providing eight replicates per treatment in each trial (n = 20 chicks/replicate). Environmental conditions in the TN group were established to simulate commercial production settings. Heat stress chickens were exposed to cyclic HS at 35 °C for 12 h/day from days 7−42. Performance parameters, intestinal permeability, bone parameters, meat quality, and leukocyte proportions were estimated. There was a significant (p < 0.05) reduction in body weight (BW), BW gain, and feed intake, but the feed conversion ratio increased in chickens under cyclic HS. Moreover, HS chickens had a significantly higher gut permeability, monocyte and basophil levels, but less bone mineralization than TN chickens. Nevertheless, the TN group had significant increases in breast yield, woody breast, and white striping in breast fillets compared to HS. These results present an alternative model to our previously published continuous HS model to better reflect commercial conditions to evaluate commercially available nutraceuticals or products with claims of reducing the severity of heat stress.

Neuropeptide Y (NPY) is one of the most abundant and ubiquitously expressed neuropeptides in both the central and peripheral nervous systems, and its regulatory effects on feed intake and appetite- have been extensively studied in a wide variety of animals, including mammalian and non-mammalian species. Indeed, NPY has been shown to be involved in the regulation of feed intake and energy homeostasis by exerting stimulatory effects on appetite and feeding behavior in several species including chickens, rabbits, rats and mouse. More recent studies have shown that this neuropeptide and its receptors are expressed in various peripheral tissues, including the thyroid, heart, spleen, adrenal glands, white adipose tissue, muscle and bone. Although well researched centrally, studies investigating the distribution and function of peripherally expressed NPY in avian (non-mammalian vertebrates) species are very limited. Thus, peripherally expressed NPY merits more consideration and further in-depth exploration to fully elucidate its functions, especially in non-mammalian species. The aim of the current review is to provide an integrated synopsis of both centrally and peripherally expressed NPY, with a special focus on the distribution and function of the latter.

Heat stress has strong adverse effects on poultry production and, thereby, threats its sustainability, which energized scientists to search for innovative and effective solutions. Here, we undertook this study to evaluate the effects of in-feed herbal adaptogen (stress response modifier) supplementation on growth performances, meat quality, and breast amino acid profile in chronic cyclic heat-stressed broilers. Day-old male Cobb 500 chicks ( n = 720) were randomly assigned, in environmental chambers ( n = 12, 24 pens), to three diet-treatments: a three-phase corn-soybean based diet fed as such (Control, C), or supplemented with the herbal adaptogen at 500 g/1000 kg control diet (NR-PHY-500) or at 1 kg/1000 kg control diet (NR-PHY-1000). From d29 to d42, birds from 9 chambers were exposed to cyclic heat stress (HS, 35°C from 9:30 am-5:30 pm), however, the rest of the chamber were maintained at thermoneutral conditions (24°C, TN), which creates 4 experimental groups: C-TN, C-HS, NR-PHY-500HS, and NR-PHY-1000HS (6 pens/group, 168 birds/group). HS altered growth performance via depression of feed intake and body weight. Adaptogen supplementation stimulated feed intake and averaged 65.95 and 83.25 g better body weight and 5 and 10 points better FCR at low and high dose, respectively, compared to heat-stressed birds. This increase in body weight was mirrored in enhanced weights of body parts (breast, tender, wings, and legs). Adaptogen supplementation modulated also breast amino acid profile, pH, color, and quality. Together, these data suggested that adaptogen supplementation could be a promising solution to alleviate heat stress, however further in-depth investigation for its mode of action and its underlying mechanisms are warranted., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Greene, Maynard, Owens, Meullenet and Dridi.)

Heat stress (HS) has been reported to disrupt nutrient digestion and absorption in broilers. These effects may be more prominent in fast-growing chickens due to their high metabolic activity. However, the underlying molecular mechanisms are not yet fully elucidated. Hence, the current study aimed to evaluate the effect of chronic HS on jejunal nutrient transport in slow- (Athens Canadian Random Bred, ACRB from 1950), moderate- (The 1995 random bred, 95RAN), rapid- (modern broilers, modern random bred, MRB) growing birds and their ancestor wild jungle fowl (JF). One-day male chicks ( n = 150/line) were placed by line in environmentally controlled chambers and kept under the same industry-standard environmental conditions until d28. On d29, an 8-h daily cyclic HS (36°C) was applied to half of the chambers, which lasted until d55, while keeping the rest under thermal neutral (TN, 24°C) conditions. Jejunum tissues were collected for morphology assessment and molecular analysis of carbohydrate-, amino acid-, and fatty acid-transporters. MRB exhibited the highest body weight (BW) followed by 95RAN under both conditions. HS decreased feed intake (FI) in MRB and 95RAN, which resulted in lower BW compared to their TN counterparts; however, no effect was observed in ACRB and JF. MRB showed a greater villus height (VH) to crypt depth (CD) ratio under both environmental conditions. Molecular analyses showed that glucose transporter (GLUT) 2, 5, 10, and 11 were upregulated in MRB compared to some of the other populations under TN conditions. HS downregulated GLUT2, 10, 11, and 12 in MRB while it increased the expression of GLUT1, 5, 10, and 11 in JF. GLUT2 protein expression was higher in JF compared to ACRB and MRB under TN conditions. It also showed an increase in ACRB but no effect on 95RAN and MRB under HS conditions. ACRB exhibited greater expression of the EAAT3 gene as compared to the rest of the populations maintained under TN conditions. HS exposure did not alter the gene expression of amino acid transporters in MRB. Gene expression of CD36 and FABP2 was upregulated in HS JF birds. Protein expression of CD36 was downregulated in HS JF while no effect was observed in ACRB, 95RAN, and MRB. Taken together, these data are the first to show the effect of HS on jejunal expression of nutrient transporters in three broiler populations known to represent 70 years of genetic progress in the poultry industry and a Red Jungle Fowl population representative of the primary ancestor of domestic chickens., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Abdelli, Ramser, Greene, Beer, Tabler, Orlowski, Pérez, Solà-Oriol, Anthony and Dridi.)

Neuropeptide Y (NPY) is a highly conserved 36-amino acid neurotransmitter, which is primarily expressed in the mammalian arcuate nucleus of the hypothalamus. It is a potent orexigenic neuropeptide, stimulating appetite and inducing feed intake in a variety of species. Recent research has shown that NPY and its receptors can be expressed by peripheral tissues, but their role is not yet well defined. Specifically, this information is particularly sparse in avian species. Therefore, the aim of this study was to determine the expression of NPY and its receptors, and determine their regulation by environmental and nutritional stressors, in the skeletal muscle of avian species using in vivo and in vitro approaches. Here, we show that NPY and its receptors are expressed in chicken breast and leg muscle as well as in quail myoblast (QM7) cell line. Intraperitoneal injection of recombinant NPY increased feed intake in 9-d old chicks and upregulated the expression of NPY and NPY receptors in breast and leg muscle, suggesting autocrine and/or paracrine roles for NPY. Additionally, NPY is able to modulate the mitochondrial network. In breast muscle, a low dose of NPY upregulated (P < 0.05) the expression of genes involved in ATP production (uncoupling protein, UCP; nuclear factor erythroid 2 like 2, NFE2L2) and dynamics (mitofusin 1, MFN1), while a high dose decreased (P < 0.05) markers of mitochondrial dynamics (mitofusin 2, MFN2; OPA1 mitochondrial dynamin like GTPase, OPA1) and increased (P < 0.05) genes involved in mitochondrial biogenesis (D-loop, peroxisome proliferator activated receptor gamma, PPARG). In leg muscle, NPY decreased (P < 0.05) markers of mitochondrial biogenesis and ATP synthesis (D-loop; peroxisome proliferator activated receptor alpha, PCG1A; peroxisome proliferator-activated receptor gamma, coactivator 1 beta, PPARGC1B; PPARG; NFE2L2). In QM7 cells, genes associated with mitochondrial biogenesis, dynamics, and ATP synthesis were all upregulated (P < 0.05), even though basal respiration and ATP production were decreased (P < 0.05) with NPY treatment as measured by XF Flux analysis. Together, these data show that the NPY system is expressed in avian skeletal muscle and plays a role in mitochondrial function., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Muscle development during embryonic and early post-hatch growth is primarily through hyperplastic growth and accumulation of nuclei through satellite cell contribution. Post-hatch, muscle development transitions from hyperplasia to hypertrophic growth of muscle fibers. Commercial selection for breast yield traditionally occurs at ages targeting hypertrophic rather than hyperplastic growth. This has resulted in the production of giant fibers and concomitant challenges with regard to muscle myopathies. The current study investigates the impact of selection during the period of hyperplastic growth. It is hypothesized that selection for percentage breast yield during hyperplasia will result in an increased number of muscle cells at hatch and potentially impact muscle fiber characteristics at processing. This study characterizes the breast muscle histology of three broiler lines at various ages in the growth period. The lines include a random bred control (RAN) as well as lines which have been selected from RAN for high (HBY4) and low (LBY4) percentage 4-day breast yield. Post-rigor pectoralis major samples from six males of each line and age were collected and stored in formalin. The sample ages included embryonic day 18 (E18), post-hatch day 4 (d4), and day 56 (d56). The samples were processed using a Leica tissue processor, embedded in paraffin wax, sectioned, and placed on slides. Slides were stained using hematoxylin and eosin. E18 and d4 post-hatch analysis showed advanced muscle fiber formation for HBY4 and immature muscle development for LBY4 as compared to RAN. Post-hatch d56 samples were analyzed for fiber number, fiber diameter, endomysium, and perimysium spacing. Line HBY4 had the largest muscle fiber diameter (54.2 ± 0.96 μm) when compared to LBY4 (45.4 ± 0.96 μm). There was no line difference in endomysium spacing while perimysium spacing was higher for HBY4 males. Selection for percentage 4-day breast yield has impacted the rate and extent of muscle fiber formation in both the LBY4 and HBY4 lines with no negative impact on fiber spacing. The shift in processing age to later ages has exposed issues associated with muscle fiber viability. Selection during the period of muscle hyperplasia may impact growth rate; however, the potential benefits of additional satellite cells are still unclear., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Orlowski, Dridi, Greene, Coy, Velleman and Anthony.)

The aim of this study was to see how spray-dried plasma (SDP) supplementation affected broiler chicken performance, intestinal permeability, and bone strength during persistent heat stress. One-day-old chicks ( n = 480) were randomly assigned into twelve environmental corrals; four thermoneutral (TN-negative control, maintained at 24 °C from d 21-42); four heat stress (HS, exposed to 35 °C from d 21-42); and four heat stress treated with 2% SDP in the feed until d 28 followed by 1% SDP until d 42 (HS-SDP). The performance and serum levels of fluorescein isothiocyanate-dextran (FITC-d) were evaluated at d 21, 28, 35, and 42. The tibias strength was evaluated on d 21 and 42. The increment in chicken temperature ( p < 0.05) was observed two h following the increase in environmental temperature in both HS groups and was associated with decreased performance parameters compared with the TN group. At d 42 of age, the chickens exposed to HS had an impaired gut permeability and decreased tibia strength compared to the TN group ( p < 0.05). However, partially feeding SDP mitigated these adverse effects significantly. These findings imply that using SDP strategically during stressful times, such as prolonged heat stress, may help mitigate its negative consequences.

This study was conducted to distinguish the effects of heat stress (HS) and feed intake (FI) on broiler chicken's physiological responses. Day-old male Cobb 500 broilers ( n = 672) were allocated to three treatments: (1) control (CTL): birds raised under normal temperature (23°C) from day 29 to 42; (2) cyclic heat stress (CHS): birds exposed to high temperatures (8 h/day at 35°C; from 9:30 am to 5:30 pm) from day 29 to 42; (3) pair-fed (PF): birds raised under thermoneutral condition but fed the same amount of feed as CHS from day 29 to 42. On day 42, 15 birds/pen were processed, to measure carcass and meat yields. To measure blood parameters and gut integrity (using fluorescein isothiocyanate-dextran), on day 42, CHS birds were sampled before (Pre-CHS) and 2 h after (Post-CHS) the temperature increased. Furthermore, after sampling CTL birds, they were exposed to 2h heat and sampled (acute heat stress, AHS). Data were analyzed using one-way ANOVA (JMP Pro15) and significance between treatments identified by LSD ( P < 0.05). BW and relative carcass yield were significantly higher in CTL compared to CHS and PF. Compared to CHS, PF had significantly higher BW and lower relative carcass yield. Breast yield was significantly higher for CTL and PF, while leg quarters and wings yield were significantly lower compared to CHS. Gut barrier integrity was significantly altered in Post-CHS and AHS compared to CTL. mRNA abundances of tumor necrosis factor-α, C-C motif chemokine ligand-20, heat shock protein ( HSP ) -27 , and HSP70 were significantly higher in Post-CHS and AHS compared to CTL. AHS had significantly higher mRNA abundances of CARD domain containing ( NLRC ) -3 and NLRC5 inflammasomes, and lower superoxide dismutase ( SOD ) -1 and SOD2 abundance compared with CTL. PF had significantly higher liver weight (% BW) compared to all other groups; while abdominal fat was significantly higher in Pre-CHS compared to CTL, PF, and AHS. Together, these data indicate that the negative effects of HS are partially due to reduced FI. However, the negative effect of HS on gut integrity, average daily gain, feed conversion ratio, and meat yield are direct and independent of the reduced FI during the HS. Thus, warrant investigating the underlying mechanisms in future research., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Emami, Greene, Kogut and Dridi.)

Injection safety is essential to reduce the risk of healthcare-associated infections (HAIs) when accessing vascular catheters. This general review evaluates the contamination of vascular catheter access ports and associated HAIs in acute care settings, focusing on open lumen stopcocks (OLSs) and disinfectable needleless closed connectors (DNCCs). PubMed was searched from January 2000 to February 2021. OLS intraluminal surfaces are frequently contaminated during patient care, increasing the risk of HAIs, and neither an isopropyl alcohol (IPA) pad nor a port-scrub device can reduce contamination effectively. In contrast, DNCCs can be disinfected, with most studies indicating less intraluminal contamination than OLSs and some studies showing decreased HAIs. While the optimal DNCC design to reduce HAIs needs to be determined, DNCCs alone or stopcocks with a DNCC bonded to the injection port should replace routine use of OLSs, with OLSs restricted to use in sterile fields. Compliance with disinfection is essential immediately before use of a DNCC as use of a non-disinfected DNCC can have equivalent or greater risk of HAIs compared with use of an OLS. The recommendations for access port disinfection in selected national and international guidelines vary. When comparing in-vitro studies, clinical studies and published guidelines, consensus is lacking; therefore, additional studies are needed, including large randomized controlled trials. IPA caps disinfect DNCCs passively, eliminate scrubbing and provide a contamination barrier; however, their use in neonates has been questioned. Further study is needed to determine whether IPA caps are more efficacious than scrubbing with disinfectant to decrease HAIs related to use of central venous, peripheral venous and arterial catheters., (Copyright © 2021 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

Woody breast (WB) condition has created a variety of challenges for the global poultry industry. To date, there are no effective treatments or preventative measures due to its unknown (undefined) etiology. Several potential mechanisms including oxidative stress, fiber-type switching, cellular damage, and altered intracellular calcium levels have been proposed to play a key role in the progression of the WB myopathy. In a previous study, we have shown that WB is associated with hypoxia-like status and dysregulated oxygen homeostasis. As satellite cells (SC) play a pivotal role in muscle fiber repair and remodeling under stress conditions, we undertook the present study to determine satellite cell fate in WB-affected birds when reared in either normoxic or hypoxic conditions. Modern random bred broilers from 2015 (n = 200) were wing banded and reared under standard brooding practices for the first 2 wk post-hatch. At 15 d, chicks were divided in 2 body weight-matched groups and reared to 6 wk in either control local altitude or hypobaric chambers with simulated altitude of 6,000 ft. Birds were provided ad libitum access to water and feed, according to the Cobb recommendations. At 6 wk of age, birds were processed and scored for WB, and breast samples were collected from WB-affected and unaffected birds for molecular analyses (n = 10/group). SCs were isolated from normal breast muscle, cultured in vitro, and exposed to normoxia or hypoxia for 2 h. The expression of target genes was determined by qPCR using 2 -∆∆Ct method. Protein distribution and expression were determined by immunofluorescence staining and immunoblot, respectively. Data were analyzed by the Student's t test with significance set at P < 0.05. Multiple satellite cell markers, myogenic factor (Myf)-5 and paired box (PAX)-7 were significantly decreased at the mRNA and protein levels in the breast muscle from WB-affected birds compared to their unaffected counterparts. Lipogenic-and adipogenic-associated factors (acetyl-CoA carboxylase, ACCα; fatty acid synthase, FASN, malic enzyme, ME; and ATP citrate lyase, ACLY) were activated in WB-affected birds. These data were supported by an in vitro study where hypoxia decreased the expression of Myf5 and Pax7, and increased that of ACCα, FASN, ME, and ACLY. Together, these data indicate that under hypoxic condition, SC change fate by switching from a myogenic to an adipogenic program, which explains at least partly, the etiology of the WB myopathy., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Proteins are considered the most expensive nutrients in commercial modern broiler production, and their dietary inclusion at low levels is pivotal to minimize feed costs and reduce nitrogen waste. The quest for an environmentally friendly source of proteins that favor the formulation of low protein diets without compromising broiler health, welfare, and growth performance has become a hotspot in nutrition research. Due to its high protein content, the naturally growing Spirulina microalgae is considered a promising nutrient source. The purpose of the present study was, therefore, to determine the effects of Spirulina supplementation on liver bacterial translocation, hematological profile, and circulating inflammatory and redox markers in broilers fed a low-protein diet. One-day-old Ross 708 male broilers ( n = 180) were randomly assigned into one of three experimental treatments: standard diet as a control, low protein diet, and low protein diet supplemented with 100 g/kg of Spirulina . Target molecular markers were measured in the peripheral blood circulation using real-time quantitative PCR. Reducing dietary proteins increased bacterial translocation and systemic inflammation as indicated by proportions of basophils among blood leukocytes. The expression levels of circulating pro-inflammatory cytokines [interleukin (IL)-3, IL-6, IL-4, IL-18, and tumor necrosis factor-α], chemokines (CCL-20), and NOD-like receptor family pyrin domain containing 3 inflammasome were significantly upregulated in birds fed the low protein diet compared with the control. The inclusion of Spirulina reversed these effects, which indicates that Spirulina reduces systemic inflammation- and bacterial translocation-induced by a low protein diet and could be a promising alternative protein source in poultry diets., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Mullenix, Greene, Emami, Tellez-Isaias, Bottje, Erf, Kidd and Dridi.)

The objective of the present research was to assess the dietary supplementation of three formulations of essential oils (EO) in chickens under heat stress (HS). Day-of-hatch Cobb 500 chicks ( n = 500) were randomly distributed into four groups: 1. HS control + control diets; 2. HS + control diets supplemented with 37 ppm EO of Lippia origanoides (LO); 3. HS + control diets supplemented with 45 ppm LO + 45 ppm EO of Rosmarinus officinalis (RO) + 300 ppm red beetroot; 4. HS + 45 ppm LO + 45 ppm RO + 300 ppm natural betaine. Chickens that received the EO showed significant ( p < 0.05) improvement on BW, BWG, FI, and FCR compared to control HS chickens. Average body core temperature in group 3 and group 4 was significantly ( p < 0.05) reduced compared with the HS control group and group 2. Experimental groups showed a significant reduction in FITC-d at 42 days, a significant increase in SOD at both days but a significant reduction of IFN-γ and IgA compared with HS control ( p < 0.05). Bone mineralization was significantly improved by EO treatments ( p < 0.05). Together these data suggest that supplemental dietary EO may reduce the harmful effects of HS.

Heat stress (HS) is a critical concern to the poultry industry as it affects both productivity and well-being. Various managerial and nutritional strategies have been proposed to mitigate the negative effects of HS in chickens, with plant-based additives showing promise. Recently, we reported the positive effect of a phytogenic feed additive (PFA) on growth performance in HS birds. Owing to the antioxidant nature of these compounds, we sought to further explore the effect of PFA on whole blood circulating chemokines, cytokines, and inflammasomes in HS broilers. Broilers (600 males, 1 d) were randomly assigned to 12 environmental chambers, subjected to 2 environmental conditions (12 h cyclic heat stress, HS, 35°C vs. thermoneutral condition [TN], 24°C) and fed 3 diets (control, PFA-C 250 ppm, PFA-C 400 ppm) in a 2 × 3 factorial design. After 21 d of cyclic HS, blood samples were collected for target gene expression analysis. HS upregulated the expression of superoxide dismutase 1 (SOD1) and downregulated glutathione peroxidase-3 (GPX-3), and there was diet × temperature interaction for SOD2, GPX-1, and GPX-3, where gene expression was increased by PFA-C250 during HS but was unchanged for PFA-C400. Plasma total antioxidant capacity (TAC) and malondialdehyde (MDA) content were increased by HS. Gene expression of interleukin-18 (IL-18) was decreased by HS, without further effect of PFA. HS increased tumor necrosis factor α (TNFα), but this effect was mitigated by PFA-C400. C-C motif chemokine ligands 4 and 20 (CCL4 and CCL20) showed a similar pattern to TNFα, with PFA-C400 ameliorating the negative effect of HS. The nucleotide-binding, leucine-rich repeat and pyrin domain containing 3 (NLRP3) inflammasome was decreased by HS and further lowered by PFA-C400, but the nucleotide-binding oligomerization domain, leucine-rich repeat, and CARD domain containing 3 (NLRC3) and nucleotide-binding, leucine-rich repeat containing X1 (NLRX1) inflammasomes were increased by PFA under TN conditions, with no effects of HS. Heat shock proteins (HSP) and heat shock factors (HSF) were unaffected by PFA or HS. Together these data indicate that gene expression of circulating inflammatory factors are dysregulated during HS, and supplemental dietary PFA may be protective., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Identification of alternatives to antibiotics in livestock and poultry is necessary. Fueled by consumer preferences, phytogenic feed additives are increasingly used in the food system; however, their mode of action is not well defined. Here, we used broiler chickens, in which appetite and feeding behavior regulation are controlled by complex mechanisms, to determine the effect of the phytogenic feed additive "comfort" (PFA-C) as well as its underlying molecular mechanisms on growth performance in heat-stressed broiler chickens. Heat stress significantly increased birds' core body temperature, water intake, and the hypothalamic expression of heat shock protein (HSP) 70, whereas it decreased feed intake, BW, and woody breast incidence. Phytogenic feed additive "comfort" supplementation downregulated the hypothalamic expression of HSP70, reduced core body temperature, increased feed and water intake, and improved BW in HS broilers. At molecular levels, the effect of PFA-C on growth performance seemed to be mediated by modulation of hypothalamic expression of melanocortin receptor 2, arginine vasopressin, aquaporin 2, and sodium and potassium-transporting ATPase subunit beta 1 polypeptides. In summary, PFA-C supplementation ameliorates heat stress productivity losses via a potential cytoprotective effect, reduction of hypothalamic intracellular stress, and modulation of hypothalamic feeding- and drinking-related polypeptide expression., (Published by Elsevier Inc.)

Orexins are originally characterized as orexigenic hypothalamic neuropeptides in mammals. Subsequent studies found orexin to be expressed and perform pleiotropic functions in multiple tissues in mammals. In avian (non-mammalian) species, however, orexin seemed to not affect feeding behavior and its physiological roles are poorly understood. Here, we provide evidence that orexin and its related receptors are expressed in chicken hepatocytes. Double immunofluorescence staining showed that orexin is localized in the ER, Golgi, and in the lysosomes in LMH cells. Brefeldin A treatment reduced orexin levels in the culture media, but increased it in the cell lysates. Administration of recombinant orexins upregulated the expression of orexin system in the liver of 9-day old chicks, but did not affect feed intake. Recombinant orexins increased fatty acid synthase (FASN) protein levels in chicken liver, activated acetyl-CoA carboxylase (ACCα), and increased FASN, ATP citrate lyase(ACLY), and malic enzyme (ME) protein expression in LMH cells. Blockade ERK1/2 activation by PD98059 attenuated these stimulating effects of orexin on lipogenic factors. Overexpression of ERK1/2 increased the expression of lipogenic genes, and orexin treatment induced the phosphorylated levels of ERK1/2 Thr202/Tyr204 , but not that of p38 Thr180/Tyr182 or JNK1/2 Thr183/Tyr185 in chicken liver and LMH cells. Taken together, this is the first report evidencing that orexin is expressed and secreted from chicken hepatocytes, and that orexin induced hepatic lipogenesis via activation of ERK1/2 signaling pathway.

Woody breast (WB) myopathy is a major concern and economic burden to the poultry industry, and for which, there is no effective solution because of its unknown etiology. In a previous study, we have shown that phytase (Quantum Blue, QB) reduces the WB severity by 5% via modulation of oxygen homeostasis-related pathways. As WB has been suggested to be associated with lipid dysmetabolism, we aimed to determine the effect of QB on WB and breast muscle fatty acid profile. Male broilers were subjected to 6 treatments (96 birds/treatment): a nutrient adequate control group (PC), the PC supplemented with 0.3% myo-inositol (PC + MI), a negative control (NC) deficient in available P and Ca by 0.15 and 0.16%, respectively, the NC fed with QB at 500 (NC+500 FTU), and 1,000 (NC+ 1,000 FTU) or 2,000 FTU/kg of feed (NC+2,000 FTU). Woody breast and white striping scores were recorded, and fatty acid profiles were determined using gas liquid chromatography. Woody breast-affected muscles exhibited a significant higher incidence of white striping as liquid chromatography analysis reveals an imbalance of fatty acid profile in the breast of WB-affected birds with a significant higher percent of saturated fatty acids (SFA, myristic [14:0], pentadecanoic [15:0], and margaric [17:0]) and monounsaturated fatty acids (myristoleic [14:1], palmitoleic [16:1c], 10-trans-heptadecenoic [17:1t], oleic [18:1c9], and vaccenic [18:1c11]), and lower content of polyunsaturated fatty acids (PUFA) and omega-3 (P < 0.05). Quantum Blue at high doses (1,000 and 2,000 FTU) significantly reduces the percent of SFA and increases that of PUFA compared with the control group. In conclusion, WB myopathy seemed to be associated with an imbalance of fatty acid profile, and QB ameliorates the severity of WB potentially via modulation of SFA and PUFA contents., (Copyright © 2020. Published by Elsevier Inc.)

As a result of genetic selection, the modern broiler is more efficient, higher yielding, and faster growing than the bird of the 1950s. Unfortunately, as a result of improvement in growth rate, the modern broiler has the potential to struggle under heat stress conditions. The present study evaluates 3 different random bred populations and a common ancestor under both a thermal neutral and heat stress conditions after a 54-D grow-out period. The lines used in this study included the Athens Canadian Random Bred (ACRB), a 1995 Random Bred (95RAN), a 2015 Random Bred (MRB), and a Junglefowl (JF). Male chicks (n = 150/line) were placed by line in environmentally controlled chambers. An 8-h daily cyclic heat stress (36°C) was applied to half of the chambers beginning on day 28 (HS) and lasting until processing at day 55, while the remaining chambers remained thermal neutral (TN) at 26°C. Dock weights and carcass weights were lower in the HS-95RAN and HS-MRB, compared to their TN counterparts, while the ACRB and JF had no difference in dock and carcass weights regardless of environmental condition. The MRB line had the highest breast yield (27.79%) while the JF (12.79%) and ACRB (12.42%) had the lowest. The 95RAN line had the highest abdominal fat percentage (2.83%) while the MRB line had the lowest moisture uptake during chill. The HS exposure lowered overall breast yield and breast pH at 15 min and 4 h postmortem but did not have an impact on color (L∗) or 24 h breast drip loss. The MRB was scored for both woody breast and white striping. The TN-MRB group had a higher incidence of moderate and severe woody breast and white striping than the HS-MRB group. Based on the results of this study, it appears that HS has a greater negative impact on the higher yielding lines (MRB and 95RAN) than the ACRB and JF and that clear line differences exist between the random bred lines and their common ancestor., (Copyright © 2020. Published by Elsevier Inc.)

High environmental temperature has strong adverse effects on poultry production, welfare, and sustainability and, thereby, constitutes one of the most challenging stressors. Although colossal information has been published on the effects of heat stress on poultry productivity and gut health, the fundamemntal mechanisms associated with heat stress responses and intestinal barrier function are still not well defined. The aim of the present study was, therefore, to determine the effects of acute (2 h) heat stress on growth performance, gut integrity, and intestinal expression of heat shock and tight junction proteins in slow- (broilers of the 1950's, ACRB), moderate- (broilers of 1990's, 95RAN), rapid-(modern broilers, MRB) growing birds, and their ancestor wild jungle fowl (JF). Heat stress exposure significantly increased the core body temperature of 95RAN and MRB chickens by ~0.5-1°C, but not that of JF and ACRB compared to their counterparts maintained at thermoneutral conditions. Heat stress also depressed feed intake and increased serum fluorescein isothiocyanate-dextran (FITC-D) levels ( P < 0.05) in modern broilers (95RAN and MRB) but not in JF and ACRB, indicating potential leaky gut syndrome. Molecular analyses showed that heat stress exposure significantly up regulated the duodenal expression of occludin ( OCLN) and lipocalin ( LCN2) in ACRB, zonula occludens ( ZO-2) , villin1 (VIL1) , and calprotectin ( CALPR) in 95 RAN, and only CALPR in MRB compared to their TN counterparts. In the jejunum however, heat stress down regulated the expression of PALS1-associated tight junction protein ( PATJ) in ACRB, 95RAN, and MRB, and that of cadherin1 ( CDH1) in MRB. In the ileum, heat stress significantly down regulated the expression of OCLN in 95 RAN, ZO-1 in MRB, gap junction protein alpha1 ( GJA1) in JF, and VIL1 in ACRB compared to their TN counterparts. In summary, this is the first report, to our knowledge, showing that tight junction protein expression is environmental-, genotype-, and intestinal segment-dependent and identifying molecular signatures, such as CDH1, CALPR , and ZO-1 , potentially involved in leaky gut syndrome-induced by heat stress in MRB., (Copyright © 2020 Tabler, Greene, Orlowski, Hiltz, Anthony and Dridi.)

Heat stress (HS) is a financial and physiological burden on the poultry industry and the mitigation of the adverse effects of HS is vital to poultry production sustainability. The purpose of this study was, therefore, to determine the effects of an amino acid-chelated trace mineral supplement on growth performance, stress and inflammatory markers, and meat quality in heat-stressed broilers. One day-old Cobb 500 male broilers (n = 480) were allocated into 12 environmental chambers (24 floor pens) and divided into two groups: one group supplemented with amino acid-chelated trace mineral in drinking water and one control group. On day 28, birds were subjected to chronic heat stress (HS, 2 wk, 35 °C and 20% to 30% RH) or maintained at thermoneutral condition (TN, 24 °C) in a 2 × 2 factorial design. Feed intake (FI), water consumption, and body weight were recorded. At day 42, serum fluorescein isothiocyanate dextran (FITC-D) levels, blood gas, electrolyte, and stress markers were measured. Jejunum samples were collected to measure gene expression of stress, inflammation, and tight junction proteins. The rest of the birds were processed to evaluate carcass traits. HS resulted in an increase in core body temperature, which increased water intake and decreased FI, body weight, and feed efficiency (P < 0.05). HS reduced carcass yield and the weight of all parts (P < 0.05). HS significantly increased levels of circulating corticosterone (CORT), heat shock protein 70 (HSP70), interleukin 18 (IL-18), tumor necrosis factor alpha, C-reactive protein, and nucleotide-binding oligomerization domain leucine-rich repeat and pyrin domain-containing 3 expression. HS significantly increased serum FITC-D levels and the expression of HSP70 and IL-18 in the jejunum. Although it did not affect the growth performance, amino acid-chelated trace mineral supplementation reversed the effect of HS by reducing CORT and FITC-D levels and the expression of stress and proinflammatory cytokines in the circulation and the jejunum. However, it upregulated these parameters in birds maintained under TN conditions. Together, these data indicate that the amino acid-chelated trace mineral might alleviate stress and inflammation and improve gut integrity in heat-stressed but not thermoneutral broilers., (© The Author(s) 2020. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The purpose of this study was to evaluate heat stress as a model to induce gastrointestinal leakage in broiler chickens. On the day of hatch, 320 chicks were allocated into 8 environmental chambers, 4 thermoneutral (TN) and 4 continuous heat stress (HS). Each chamber was divided into 2 pens containing separate feeders and water jugs (8 replicates per treatment, 20 birds/pen). The environment was established to simulate production setting as best possible for the first 21 D. A gradual reduction of temperature from 32°C to 24°C with relative humidity at 55 ± 5% was adopted for the first 21 D. At the time of HS, the HS groups were exposed to 35°C from Day 21 to 42, while thermoneutral ones were maintained at 24°C from Day 21 to 42. Chickens were equipped with a Thermochron temperature logger for continuous monitoring of core body temperature. The environmental temperature and relative humidity were continuously recorded. Fluorescein isothiocyanate-dextran (FITC-d) was orally gavaged to 2 chickens/replicate (n = 16) randomly selected on days 21, 28, 35, and 42. After 1 h of oral gavage, blood samples were collected to determine the passage of FITC-d. Tibias were removed from all chickens to evaluate break strength only on 21 D and 42 D (before HS and at the end of the trial). Performance parameters were evaluated weekly from 21 D to the end of the trial. Body temperature was significantly (P < 0.05) increased after 2 h of starting HS and remained that way until the end of the study. Chronic HS caused an increase in core body temperature which decreased feed intake, body weight, and feed efficiency (28, 35, and 42 D) when compared with control TN chickens. Similarly, serum FITC-d was significantly increased in HS chickens at all points of evaluation. Chronic HS also caused a significant reduction of bone strength at 42 D when compared with the control chickens. The results from the present study suggest that HS can be a robust model to induce gut leakage in broiler chickens., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Objective: Oxidized phospholipids (OxPL), such as the oxidized derivatives of 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine, 1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphorylcholine, and 1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphorylcholine, have been shown to be the principal biologically active components of minimally oxidized LDL (low-density lipoprotein). The role of OxPL in cardiovascular diseases is well recognized, including activation of inflammation within vascular cells. Atherosclerotic Apoe -/- mice fed a high-fat diet develop antibodies to OxPL, and hybridoma B-cell lines producing natural anti-OxPL autoantibodies have been successfully generated and characterized. However, as yet, no studies have been reported demonstrating that treatment with OxPL neutralizing antibodies can be used to prevent or reverse advanced atherosclerosis. Approach and Results: Here, using a screening against 1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphorylcholine/1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphorylcholine, we generated a novel IgM autoantibody, 10C12, from the spleens of Apoe -/- mice fed a long-term Western diet, that demonstrated potent OxPL neutralizing activity in vitro and the ability to inhibit macrophage accumulation within arteries of Apoe -/- mice fed a Western diet for 4 weeks. Of interest, 10C12 failed to inhibit atherosclerosis progression in Apoe -/- mice treated between 18 and 26 weeks of Western diet feeding likely due at least in part to high levels of endogenous anti-OxPL antibodies. However, 10C12 treatment caused a 40% decrease in lipid accumulation within aortas of secreted IgM deficient, sIgM -/- Apoe -/- , mice fed a low-fat diet, when the antibody was administrated between 32-40 weeks of age., Conclusions: Taken together, these results provide direct evidence showing that treatment with a single autoimmune anti-OxPL IgM antibody during advanced disease stages can have an atheroprotective outcome.

Poultry well-being and economic burden due to heat stress (HS) are of great importance to the poultry industry. Efficient design and effective evaluation of any strategies to alleviate the adverse effects of HS on poultry production require an accurate measurement of stress. However, current methods for monitoring stress in poultry are less than ideal, as they are invasive or subjective, and therefore variable. Here, we demonstrated that HSP70, the well-established intracellular stress chaperone, is expressed in chicken feather and is responsive to HS, and could therefore be used as a stress marker. Growing feathers were collected from the same bird (Cobb500, n = 9) in the morning (barn temperature 24°C) and afternoon (barn temperature 29.5°C) in the summer (2016 June 21) in Arkansas, USA. In the afternoon, the birds were panting and their core body temperatures were significantly higher compared to the morning time, as illustrated by iButton data thermo-loggers. Concomitantly, blood HSP70 mRNA and feather HSP70 expression (mRNA and protein) were significantly increased in the afternoon compared to the morning time. Similarly, HSP70 protein expression in the duodenum was also significantly higher in the afternoon compared to the morning period. Together, these finding identify feather HSP70 as a novel non-invasive molecular signature that mirrors the intracellular and systemic stress, which can be useful to monitor well-being of chickens and other avian species under different challenges., (© 2019 Poultry Science Association Inc.)

In humans, alterations in bone metabolism have been associated with myopathies. We postulate the hypothesis that perhaps similar pathologies can also be associated in modern chickens. Hence, this study aimed to assess the fat infiltration in bone marrow and its repercussion on broiler chicken affected by Wooden Breast (WB) myopathy. Ten Cobb 500 live birds with extreme rigidity of the Pectoralis major (PM) muscle were selected as WB affected chickens by physical examination of the muscle at 49 days of age, whereas ten chickens healthy with no physical signs of hardness in the breast muscle were considered to be unaffected. Macroscopic lesions in affected chickens included areas of firm and inflamed muscle with pale appearance, hemorrhaging, and viscous exudate on the surface. Bone marrow and sections of the PM muscle were collected and analyzed for light microscopy. Additionally, transmission electron microscopy was conducted in affected or unaffected muscle. Chickens affected with WB showed significant reductions ( P < 0.05) in femur diameter, calcium, and phosphorous percentage but increased breast weight, compression force and filet thickness when compared with non-affected chickens. Interestingly, bone marrow from WB chicken had subjectively, more abundant infiltration of adipose tissue, when compared with non-affected chickens. Histology of the Pectoralis major of birds with WB showed abundant infiltration of adipose tissue, muscle fibers degeneration with necrosis and infiltration of heterophils and mononuclear cells, connective tissue proliferation, and vasculitis. Ultrastructural changes of WB muscle revealed lack definition of bands in muscle tissue, or any normal ultrastructural anatomy such as myofibrils. The endomysium components were necrotic, and in some areas, the endomysium was notable only as a string of necrotic tissue between degraded myofibrils. The fascia appeared hypertrophied, with large areas of necrosis and myofiber without structural identity with degraded mitochondria adjacent to the disrupted muscle tissue. As far as we know, this is the first study that describes a subjective increase in adipose tissue in the bone marrow of chickens affected with WB when compared with non-affected chickens, and reduced bone mineralization.

The objective of this study was to evaluate the of intestinal permeability and liver bacterial translocation (BT) across a modern commercial broiler, a commercial broiler of 1995 genetics, and an unselected Jungle Fowl line. Modern 2015 (MB2015) broiler chicken, random bred line initiated from 1995 (RB1995), and the Giant Jungle fowl (JF). Chickens were randomly allocated to four different dietary treatments. Dietary treatments were (1) a control corn-based diet throughout the trial [corn-corn (C-C)]; (2) an early phase malnutrition diet where chicks received a rye-based diet for 10 days, and then switched to the control diet [rye-corn (R-C)]; (3) a malnutrition rye-diet that was fed throughout the trial [rye-rye (R-R)]; and (4) a late phase malnutrition diet where chicks received the control diet for 10 days, and then switched to the rye diet for the last phase [corn-rye (C-R)]. Paracellular permeability was evaluated using fluorescein isothiocyanate dextran (FITC-D). Liver BT was also evaluated. MB2015 and RB1995 consuming the rye-based diet showed increase serum levels of FITC-D when compared to the corn-fed chickens ( P < 0.05). Overall, MB2015 appeared to have higher enteric permeability than the JF. To our knowledge, this would be the first paper to evaluate the effect of compensatory growth on intestinal permeability and liver BT. Further studies to evaluate microbiome and inflammatory markers in these chicken models are currently being evaluated.

Phytogenics have been reported to improve growth performances in farm animals and are thereby considered as potential key solutions for antibiotic-free livestock nutrition. Yet, their effects on meat quality are still not well defined; therefore, the aim of this study was to determine the effects of 5 experimental phytogenic additives (3 dietary and 2 water supplements) on growth and meat quality in broilers. One-day-old broiler chicks (n = 576) were assigned to 48 floor pens and divided into 6 treatments (Control, AV/HGP/16 premix [AVHGP], Superliv concentrate premix [SCP], bacteriostatic herbal growth promotor [BHGP], AV/SSL/12 [AVSSL], and Superliv Gold [SG]) in a complete randomized design (8 pens/treatment with 12 birds/pen, and 96 birds/group). Feed intake and BW were recorded, and birds were processed at 42 d to evaluate carcass traits. Breast muscle tissues were excised to determine stress- and antioxidant-related genes expression. Both AVSSL- and SG-treated broilers produced heavier (P < 0.05) slaughter weights compared with the control-fed broilers, whereas AVSSL supplementation decreased (P < 0.05) fat pad size and increased (P < 0.05) breast weights compared with the control-fed broilers. Although pH and a* values remained unchanged, L* was decreased (P < 0.05) in all treatment and b* was reduced (P < 0.05) in SG when compared with controls. The trained sensory panelists detected more (P < 0.05) green herb flavor in the breast meat from AVHGP than SCP, SG, and control birds. The expression of superoxide dismutase 2, extracellular signal-regulated kinase 1/2, and JNK gene was upregulated in AVHGP and BHGP compared with the control (P < 0.05). Together, these results indicated that phytogenic additives might improve meat quality of broilers through modulation of stress- and antioxidant-related pathways., (© The Author(s) 2018. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)