Estimating In Vitro Protein Digestion and Protein Digestibility Corrected Amino Acid Score of Chicken Breasts Affected by White Striping and Wooden Breast Abnormalities.

An understanding regarding impacts of growth-related myopathies, i.e., white striping (WS) and wooden breast (WB), on the quality of dietary protein from cooked chicken breast is still limited. This study aimed at comparing protein content and in vitro protein digestion and estimating the in vitro protein digestibility corrected amino acid score (PDCAAS) of cooked chicken meat exhibiting different abnormality levels (i.e., normal, WS, and WS + WB). The results show that the WS + WB samples exhibited lower protein content, greater cooking loss, and greater lipid oxidation than those of normal samples (p < 0.05). No differences in protein carbonyls or the myofibril fragmentation index were found (p ≥ 0.05). Cooked samples were hydrolyzed in vitro using digestive enzyme mixtures that subsequently mimicked the enzymatic reactions in oral, gastric, and intestinal routes. The WS + WB samples exhibited greater values of free NH2 and degree of hydrolysis than the others at all digestion phases (p < 0.05), suggesting a greater proteolytic susceptibility. The in vitro PDCAAS of the WS + WB samples was greater than that of the other samples for pre-school children, school children, and adults (p < 0.05). Overall, the findings suggest that the cooked chicken breast with the WS + WB condition might provide greater protein digestibility and availability than WS and normal chicken breasts.

Impact of Wooden Breast myopathy on in vitro protein digestibility, metabolomic profile, and cell cytotoxicity of cooked chicken breast meat.

This study investigated the impacts of Wooden Breast (WB) abnormality on in vitro protein digestibility and cytotoxicity of cooked chicken breast meat. Chicken breasts without (non-WB, n = 6) or with severe WB condition (WB, n = 6) were cooked and subjected to static in vitro protein digestion. The results showed no significant differences in free-NH2, degree of hydrolysis and distribution of peptide molecular weight between non-WB and WB samples at late intestinal digestion (P5), suggesting no adverse effects of WB on protein digestibility. Based on peptidomic analysis, P5 fraction of WB showed greater content of peptides with oxidative modification than that of non-WB. Untargeted metabolomics did not find any metabolites with potential toxicity either in non-WB and WB. Hydrolyzed non-WB and WB (1.56-100 µg/mL) did not affect viability of Caco-2 and Vero cells but addition of WB samples reduced Caco-2 cell viability compared with non-WB.

Thermal impacts on transcriptome of Pectoralis major muscle collected from commercial broilers, Thai native chickens and its crossbreeds.

OBJECTIVE: The main objective of this study was to define molecular mechanisms associated with thermal stress responses of chickens from commercial broilers (BR, Ross 308), Thai native chickens (NT) and crossbreeds between BR×NT (H75). METHODS: Twenty days before reaching specific market age, chickens from each breed were divided into control and thermal-stressed groups. The stressed groups were exposed to a cyclic thermal challenge (35°C±1°C for 6 h, followed by 26°C±1°C for 18 h) for 20 days. Control group was raised under a constant temperature of 26°C±1°C. Pectoralis major (n = 4) from each group was collected for transcriptome analysis using HiSeq Illumina and analysis of glycogen and lactate. Gene expression patterns between control and thermalstressed groups were compared within the same breeds. RESULTS: Differentially expressed transcripts of 65, 59, and 246 transcripts for BR, NT, and H75, respectively, were revealed by RNA-Seq and recognized by Kyoto encyclopedia of genes and genomes database. Pathway analysis underlined altered glucose homeostasis and protein metabolisms in all breeds. The signals centered around phosphatidylinositol 3-kinase (PI3K)/Akt signaling, focal adhesion, and MAPK signaling in all breeds with slight differences in molecular signal transduction patterns among the breeds. An extensive apoptosis was underlined for BR. Roles of AMPK, MAPK signaling and regulation of actin cytoskeleton in adaptive response were suggested for H75 and NT chickens. Lower glycogen content was observed in the breast muscles of BR and NT (p<0.01) compared to their control counterparts. Only BR muscle exhibited increased lactate (p<0.01) upon exposure to the stress. CONCLUSION: The results provided a better comprehension regarding the associated biological pathways in response to the cyclic thermal stress in each breed and in chickens with different growth rates.

Development of chitin nanofiber coatings for prolonging shelf life and inhibiting bacterial growth on fresh cucumbers.

The widespread usage of petroleum-based polymers as single-use packaging has had harmful effects on the environment. Herein, we developed sustainable chitin nanofiber (ChNF) coatings that prolong the shelf life of fresh cucumbers and delay the growth of pathogenic bacteria on their surfaces. ChNFs with varying degrees of acetylation were successfully prepared via deacetylation using NaOH with treatment times of 0-480 min and defibrillated using mechanical blending. With longer deacetylation reaction times, more acetamido groups (-NHCOCH3) in chitin molecules were converted to amino groups (-NH2), which imparted antibacterial properties to the ChNFs. The ChNF morphologies were affected by deacetylation reaction time. ChNFs deacetylated for 240 min had an average width of 9.0 nm and lengths of up to several µm, whereas rod-like structured ChNFs with a mean width of 7.3 nm and an average length of 222.3 nm were obtained with the reaction time of 480 min. Furthermore, we demonstrated a standalone ChNF coating to extend the shelf life of cucumbers. In comparison to the rod-like structured ChNFs, the 120 and 240-min deacetylated ChNFs exhibited a fibril-like structure, which considerably retarded the moisture loss of cucumbers and the growth rate of bacteria on their outer surfaces during storage. Cucumbers coated with these 120 and 240-min deacetylated ChNFs demonstrated a lower weight loss rate of ⁓ 3.9% day-1 compared to the uncoated cucumbers, which exhibited a weight loss rate of 4.6% day-1. This protective effect provided by these renewable ChNFs holds promising potential to reduce food waste and the use of petroleum-based packaging materials.

Isolation and Characterization of Mannanase-Producing Bacteria for Potential Synbiotic Application in Shrimp Farming.

Prebiotics such as mannan-oligosaccharides (MOS) are a promising approach to improve performance and disease resistance in shrimp. To improve prebiotic utilization, we investigated the potential probiotics and their feasibility of synbiotic use in vitro. Two bacterial isolates, Man26 and Man122, were isolated from shrimp intestines and screened for mannanase, the enzyme for mannan digestion. The crude mannanase from both isolates showed optimal activities at pH 8 with optimum temperatures at 60 °C and 50 °C, respectively. The enzymes remained stable at pH 8−10 for 3 h (>70% relative activity). The thermostability range of Man26 was 20−40 °C for 20 min (>50%), while that of Man122 was 20−60 °C for 30 min (>50%). The Vmax of Man122 against locust bean gum substrate was 41.15 ± 12.33 U·mg−1, six times higher than that of Man26. The Km of Man26 and Man122 were 18.92 ± 4.36 mg·mL−1 and 34.53 ± 14.46 mg·mL−1, respectively. With the addition of crude enzymes, reducing sugars of copra meal, palm kernel cake, and soybean meal were significantly increased (p < 0.05), as well as protein release. The results suggest that Man26 and Man122 could potentially be used in animal feeds and synbiotically with copra meal to improve absorption and utilization of feedstuffs.

Differential expression patterns of genes associated with metabolisms, muscle growth and repair in Pectoralis major muscles of fast- and medium-growing chickens.

The aim of this study was to investigate the expression of genes related to muscle growth, hypoxia and oxidative stress responses, a multi-substrate serine/threonine-protein kinase (AMPK) and AMPK-related kinases, carbohydrate metabolism, satellite cells activities and fibro- adipogenic progenitors (FAPs) in fast-growing (FG) (n = 30) and medium-growing (MG) chickens (n = 30). Pectoralis major muscles were collected at 7d, 14d, 21d, 28d, 35d and 42d of age. According to their macroscopic features, the samples from FG up to 21d of age were classified as unaffected, while all samples collected at an older age exhibited macroscopic features ascribable to white striping and/or wooden breast abnormalities. In contrast, MG samples did not show any feature associated to muscle disorders. The absolute transcript abundance of 33 target genes was examined by droplet digital polymerase chain reaction. The results showed differential gene expression profiles between FG and MG chickens at different ages. While most genes remained unchanged in MG chickens, the expression patterns of several genes in FG were significantly affected by age. Genes encoding alpha 1, alpha 2, beta 2 and gamma 3 isoforms of AMPK, as well as AMPK-related kinases, were identified as differentially expressed between the two strains. The results support the hypothesis of oxidative stress-induced muscle damage with metabolic alterations in FG chickens. An increased expression of ANXA2, DES, LITAF, MMP14, MYF5 and TGFB1 was observed in FG strain. The results suggest the occurrence of dysregulation of FAP proliferation and differentiation occurring during muscle repair. FAPs could play an important role in defining the proliferation of connective tissue (fibrosis) and deposition of intermuscular adipose tissue which represents distinctive traits of muscle abnormalities. Overall, these findings demonstrate that dysregulated molecular processes associated with myopathic lesions in chickens are strongly influenced by growth rate, and, to some extent, by age.

In Vitro Protein Digestion of Cooked Spent Commercial Laying Hen and Commercial Broilers Breast Meat.

Chicken meat from spent laying hens (SHs) has been considered as nutritive as the meat of commercial broilers (CBs) based on chemical composition. High insoluble collagen in SH meat might reduce protein digestibility and bio-accessibility compared to CB meat. This study aimed at comparing the in vitro protein digestibility of CB and SH cooked breast meat. In the first part, CB samples were digested using two static in vitro digestion methods and collected at different digestion points for determining the degree of hydrolysis (DH). The method providing a greater DH value was chosen for comparing protein digestibility between CB and SH samples. The activities of used enzymes during in vitro digestion were evaluated based on bicinchoninic acid assay 2,4,6-trinitrobenzenesulfonic acid colorimetric method, gas chromatography-mass spectrometry, and sodium dodecyl sulfate-polyacrylamide electrophoresis. Particle size distribution of solid content collected from hydrolysate was also determined. The results showed that after digestion, CB showed 1−3 mg/mL protein concentration lower, while 7−13% DH and 50−96 µmoL/g protein-free NH2 groups higher when compared to those of SH. Based on sodium dodecyl sulfate-polyacrylamide electrophoresis, CB samples exhibited greater intensity of band at MW < 15 kDa than that of SH. Regarding particle size in terms of volume weighted mean (D[4,3]), at the end of the oral phase, the end of the gastric phase, and the beginning of the intestinal phase, D[4,3] of the SH samples were 133.17 ± 2.16, 46.52 ± 2.20, and 112.96 ± 3.63 µm, respectively, which were greater than those of CB (53.28 ± 1.23, 35.59 ± 1.19, and 51.68 ± 1.25 µm). However, at the end of the intestinal phase, D[4,3] of SH and CB, which were 17.19 ± 1.69 and 17.52 ± 2.46 µm, respectively, did not significantly differ from each other. The findings suggested a greater in vitro protein digestibility of cooked CB breast meats than that of SH ones.

Chitosan coating for the preparation of multilayer coated paper for food-contact packaging: Wettability, mechanical properties, and overall migration.

Cellulose-based paper is an alternative substitution for petroleum-based polymers for packaging applications, but its mechanical performance is poor when in contact with water. Herein, chitosan was applied on cellulose-based paper via a coating approach. The effects of chitosan coatings between none and five layers on the color properties, wettability, thermal properties, mechanical performance, and overall migration in food simulants of the paper were evaluated. After the application of chitosan, chitosan first filled cavities between cellulose fibers within a network, and the chitosan film was formed on the paper surface later. This resulted in a pronounced increase in wettability and mechanical properties associated with a loss of whiteness and an increase in yellowness of the coated paper. The chitosan-coated paper became hydrophobic with a water contact angle of 94.7 ± 2.8°, and a robust improvement of 156.4% for tensile strength and 114.8% for strain at break was observed for the paper coated with three layers of chitosan in wet conditions in comparison to the uncoated paper. A reduction in the migration of the low molecular residuals from the paper could be hindered by the chitosan coating. These enhanced features revealed that chitosan-coated paper could be used as a food-contact material.

Influences of Thermal Stress During Three Weeks Before Market Age on Histology and Expression of Genes Associated With Adipose Infiltration and Inflammation in Commercial Broilers, Native Chickens, and Crossbreeds.

The objectives of this study were to examine the effects of cyclic thermal stress on histological characteristics of breast muscle and gene expression regarding adipose infiltration and inflammation in breast muscles collected from different breeds of chickens. The birds, from commercial broilers (CB, Ross 308, 3 weeks), native (NT, 100% Thai native Chee, 9 weeks), H75 (crossbred; 75% broiler and 25% NT, 5 weeks), and H50 (crossbred; 50% broiler and 50% NT, 7 weeks), were equally assigned into control or treatment groups. The control samples were reared under a constant temperature of 26 ± 1°C, while the treatment groups were exposed to 35 ± 1°C (6 h per day). After a 20-day thermal challenge, 12 male birds per treatment group were randomly collected for determination of live body weight, breast weight, numbers of growth-related myopathies, and breast meat chemical composition. Histological lesions were evaluated in the pectoralis major muscle immediately collected within 20 min postmortem based on hematoxylin and eosin staining. The results indicated that despite interaction between thermal stress and breed effects, thermal challenge significantly reduced feed intake, live body weight, and breast weight of the birds and increased moisture content in breast meat (p < 0.05). An interaction between the two main factors was found for protein content (p < 0.05) for which control CB showed less protein than the other groups. Heat stress decreased histological scores for adipose infiltration in CB (p < 0.05), but it did not significantly influence such scores in the other groups. CB received histological scores for adipose tissue at greater extent than those for the other groups. Differential absolute abundance of CD36, FABP4, LITAF, PDGFRA, PLIN1, PPARG, POSTN, SCD1, and TGFB1 in the muscle samples well-agreed with the trend of histological scores, suggesting potential involvement of dysregulated fibro-adipogenic progenitors together with imbalanced lipid storage and utilization in the breast muscle. The findings demonstrated that the cyclic thermal challenge restricted growth performance and breast mass of the birds, but such effects attenuated infiltration of adipose tissue and inflammatory cells in the CB breast muscle.

Effects of Cyclic Thermal Stress at Later Age on Production Performance and Meat Quality of Fast-Growing, Medium-Growing and Thai Native Chickens.

The present study aimed at assessing the impact of cyclic thermal stress on production performance and meat quality of commercial broilers (BRs), Thai native chickens (NT) and the hybrids between BR and NT (H75; crossbreed 25% NT). At the age of 3, 5 and 9 weeks for BR, H75 and NT, respectively, each strain was equally divided (n = 50) into control and treatment groups. The controls were raised at a constant 26 ± 1 °C, while the treatments were subjected to thermal stress (35 ± 1 °C, 6 h daily) for 3 weeks. The results indicated that final weight and average daily gain of BR and NT treated groups were significantly lower than those of their control counterparts. Reduced body weight gain of BR and H75, as well as feed intake of H75, was observed in the treatment groups (p < 0.05). The stressed BR breasts showed decreased moisture, fat and carbohydrate, accompanied by increased protein, ash, L *-value, b*-value and shear force (p < 0.05). No significant effects (p ≥ 0.05) of the thermal stress on meat quality indices were found for H75 and NT breast samples. Pectoral myopathies were observed in BR and H75 chickens, but the numbers of cases were decreased in the thermally treated groups.

Insights Into Transcriptome Profiles Associated With Wooden Breast Myopathy in Broilers Slaughtered at the Age of 6 or 7 Weeks.

Transcriptomes associated with wooden breast (WB) were characterized in broilers at two different market ages. Breasts (Pectoralis major) were collected, 20-min postmortem, from male Ross 308 broilers slaughtered at 6 and 7 weeks of age. The breasts were classified as "non-WB" or "WB" based on palpation hardness scoring (non-WB = no abnormal hardness, WB = consistently hardened). Total RNA was isolated from 16 samples (n = 3 for 6 week non-WB, n = 3 for 6 week WB; n = 5 for 7 week non-WB, n = 5 for 7 week WB). Transcriptome was profiled using a chicken gene expression microarray with one-color hybridization technique, and compared between non-WB and WB samples of the same age. Among 6 week broilers, 910 transcripts were differentially expressed (DE) (false discovery rate, FDR < 0.05). Pathway analysis underlined metabolisms of glucose and lipids along with gap junctions, tight junction, and focal adhesion (FA) signaling as the top enriched pathways. For the 7 week broilers, 1,195 transcripts were identified (FDR < 0.05) with regulation of actin cytoskeleton, mitogen-activated protein kinase (MAPK) signaling, protein processing in endoplasmic reticulum and FA signaling highlighted as the enriched affected pathways. Absolute transcript levels of eight genes (actinin-1 - ACTN1, integrin-linked kinase - ILK, integrin subunit alpha 8 - ITGA8, integrin subunit beta 5 - ITGB5, protein tyrosine kinase 2 - PTK2, paxillin - PXN, talin 1 - TLN1, and vinculin - VCL) of FA signaling pathway were further elucidated using a droplet digital polymerase chain reaction. The results indicated that, in 6 week broilers, ITGA8 abundance in WB was greater than that of non-WB samples (p < 0.05). Concerning 7 week broilers, greater absolute levels of ACTN1, ILK, ITGA8, and TLN1, accompanied with a reduced ITGB5 were found in WB compared with non-WB (p < 0.05). Transcriptional modification of FA signaling underlined the potential of disrupted cell-cell communication that may incite aberrant molecular events in association with development of WB myopathy.

Lysine Depletion during Different Feeding Phases: Effects on Growth Performances and Meat Quality of Broiler Chickens.

The present study aimed at assessing the impact of lysine restriction performed during different feeding phases on growth performances, meat quality traits and technological properties as well as on the incidence and severity of breast muscle abnormalities. For this purpose, a total of 945 one-day-old Ross 308 male chicks was randomly divided into three experimental groups: CONT, fed a four feeding phases commercial diet, GRW I, and GRW I + II fed CONT diet with the depletion of synthetic lysine during grower I and grower I and II feeding phases, respectively. Productive performances were recorded throughout the whole rearing cycle and the incidence of breast muscle growth-related abnormalities assessed at slaughter (49 d) on 280 breasts/group. Quality traits and technological properties of breast meat were measured on a total of 54 Pectoralis major muscles. Lysine restriction only marginally affected the productive performances and the quality parameters of breast meat. The increased (p < 0.05) solubility of the protein fraction along with the remarkably higher (p < 0.05) anserine content found in GRW I + II suggests an increased energy requirement in the pectoral muscles belonging to lysine-restricted birds and supports the hypothesis of a reduced protein synthesis taking place within these muscles.

Application of One-Step Reverse Transcription Droplet Digital PCR for Dengue Virus Detection and Quantification in Clinical Specimens.

Detection and quantification of viruses in laboratory and clinical samples are standard assays in dengue virus (DENV) studies. The quantitative reverse transcription polymerase chain reaction (qRT-PCR) is considered to be the standard for DENV detection and quantification due to its high sensitivity. However, qRT-PCR offers only quantification relative to a standard curve and consists of several "in-house" components resulting in interlaboratory variations. We developed and optimized a protocol for applying one-step RT-droplet digital PCR (RT-ddPCR) for DENV detection and quantification. The lower limit of detection (LLOD95) and the lower limit of quantification (LLOQ) for RT-ddPCR were estimated to be 1.851 log10-copies/reaction and 2.337 log10-copies/reaction, respectively. The sensitivity of RT-ddPCR was found to be superior to qRT-PCR (94.87% vs. 90.38%, p = 0.039) while no false positives were detected. Quantification of DENV in clinical samples was independently performed in three laboratories showing interlaboratory variations with biases <0.5 log10-copies/mL. The RT-ddPCR protocol presented here could help harmonize DENV quantification results and improve findings in the field such as identifying a DENV titer threshold correlating with disease severity.

Influence of non-phosphate and low-sodium salt marination in combination with tumbling process on properties of chicken breast meat affected by white striping abnormality.

This study investigated the effects of non-phosphate and low-sodium (NPLS) marination on properties of white striping chicken breasts (WSCB). Chicken breasts were collected from slaughterhouse and classified as normal (NCB, n = 24) and severe WS (WSCB, n = 120). Sixty WSCB samples were vacuum-tumbled (30 min, 2 °C) with NPLS solution, containing 2.8% (w/v) potassium bicarbonate, 2.9% (w/v) potassium chloride, and 1.5% (w/v) sorbitol at the ratio of meat-to-marinade of 4 to 1 (w/w). The other 60 WSCB received no marination were assigned as nonmarinated WSCB. Properties of marinated (n = 12) and nonmarinated (n = 12) WSCB samples were determined at 0, 3, 7, 10, and 14 days of the storage at 4 °C. Properties of the NCB were also determined on day 0. Concerning day 0, the marinated WSCB exhibited higher (p < 0.05) pH, moisture content, total cooked yield, protein solubility, hardness, cohesiveness, and chewiness along with lower (p < 0.05) cooked loss, expressible water, and shear force than those of nonmarinated WSCB and NCB. Based on nuclear magnetic resonance spectroscopy, bound, intra-myofibrillar, and extra-myofibrillar water of cooked marinated WSCB were greater (p < 0.05) than those of cooked nonmarinated WSCB. The greater (p < 0.05) weight loss, moisture content, and total cooking yield were observed in marinated samples compared to those of nonmarinated WSCB throughout the storage period. Although microbial stability was reduced (p < 0.05), no difference (p ≥ 0.05) in lipid oxidation was detected between the treatments. The findings suggest the NPLS marination as a promising process for improving water holding capacity of the WSCB. PRACTICAL APPLICATION: This study presents the promising application of non-phosphate, low-sodium (NPLS) marination combined with vacuum-tumbling in improving water holding capacity of chicken breast meat affected with white striping condition. Although microbial stability of the marinated breast was negatively affected, no adverse impacts on lipid oxidation was observed during storage up to 14 days.

Nutritional Properties and Oxidative Indices of Broiler Breast Meat Affected by Wooden Breast Abnormality.

Wooden breast (WB) abnormality adversely impacts the quality of chicken meat and has been linked with oxidative stress. In this study, breast samples were taken from carcasses of 7-week-old Ross 308 broilers 20-min and 24-h postmortem. Five WB and seven non-WB control samples were assigned based on palpatory hardness (non-WB = no unusual characteristics and WB = focal or diffused hardness). WB exhibited lower contents of protein and the amino acids, i.e., isoleucine, leucine and valine, lighter surface color, lower shear force, greater drip loss and altered mineral profiles (p ≤ 0.05). Despite no difference in lipid oxidation, a greater degree of protein oxidation was found in the WB meat (p ≤ 0.05). Absolute transcript abundances of superoxide dismutase, hypoxia inducible factor 1 alpha and pyruvate dehydrogenase kinase 1 were greater in WB (p ≤ 0.05), whereas lactate dehydrogenase A expression was lower in WB (p ≤ 0.05). The findings support an association between oxidative stress and the altered nutritional and technological properties of chicken meat in WB.

Knockdown of Death-Associated Protein Expression Induces Global Transcriptome Changes in Proliferating and Differentiating Muscle Satellite Cells.

Death-associated protein (DAP) undergoes substantial changes in expression during turkey skeletal muscle development, decreasing from the 18 day embryonic stage to 1 day posthatch, and again from 1 day posthatch to 16 weeks of age. These changes suggest that DAP plays an important role at critical stages of the developmental process. The objective of this study was to elucidate the role of DAP in muscle development by examining the effect of reduced DAP expression on global gene expression in proliferating and differentiating turkey pectoralis major muscle satellite cells. Small interfering RNA was used to knock down expression of DAP and the transcriptome was subsequently profiled using a turkey skeletal muscle long oligonucleotide microarray. Microarray data were corroborated using quantitative real-time PCR. In proliferating cells, 458 loci, resulting in 378 uniquely annotated genes, showed differential expression (false discovery rate, FDR < 0.05). Pathway analysis highlighted altered eukaryotic translational initiation factors (eIFs) signaling, protein ubiquitination, sirtuin signaling, and mechanistic target of rapamycin (mTOR) signaling as the primary pathways affected in the knockdown proliferating cells. The findings underpinned the potential DAP involvement in cell proliferation of turkey satellite cells through the coordination between protein synthesis and cell cycle. In differentiating cells, 270 loci, accounting for 189 unique genes, showed differential expression (FDR < 0.05). Decreased expression of genes encoding various myofibrillar proteins and proteins involved in sarcoplasmic reticulum calcium flux suggests that DAP may affect regulation of calcium homeostasis and cytoskeleton signaling. This study provides the first evidence that reduced expression of DAP significantly alters the transcriptome profile of pectoralis major muscle satellite cells, thereby reducing proliferation and differentiation.

Transcriptional Profiles of Skeletal Muscle Associated With Increasing Severity of White Striping in Commercial Broilers.

Development of the white striping (WS) abnormality adversely impacts overall quality of broiler breast meat. Its etiology remains unclear. This study aimed at exploring transcriptional profiles of broiler skeletal muscles exhibiting different WS severity to elucidate molecular mechanisms underlying the development and progression of WS. Total RNA was isolated from pectoralis major of male 7-week-old Ross 308 broilers. The samples were classified as mild (n = 6), moderate (n = 6), or severe (n = 4), based on number and thickness of the white striations on the meat surface. The transcriptome was profiled using a chicken gene expression microarray with one-color hybridization technique. Gene expression patterns of each WS severity level were compared against each other; hence, there were three comparisons: moderate vs. mild (C1), severe vs. moderate (C2), and severe vs. mild (C3). Differentially expressed genes (DEGs) were identified using the combined criteria of false discovery rate ≤ 0.05 and absolute fold change ≥1.2. Differential expression of 91, 136, and 294 transcripts were identified in C1, C2, and C3, respectively. There were no DEGs in common among the three comparisons. Based on pathway analysis, the enriched pathways of C1 were related with impaired homeostasis of macronutrients and small biochemical molecules with disrupted Ca2+-related pathways. Decreased abundance of the period circadian regulator suggested the shifted circadian phase when moderate WS developed. The enriched pathways uniquely obtained in C2 were RNA degradation, Ras signaling, cellular senescence, axon guidance, and salivary secretion. The DEGs identified in those pathways might play crucial roles in regulating cellular ion balances and cell-cycle arrest. In C3, the pathways responsible for phosphatidylinositol 3-kinase-Akt signaling, p53 activation, apoptosis, and hypoxia-induced processes were modified. Additionally, pathways associated with a variety of diseases with the DEGs involved in regulation of [Ca2+], collagen formation, microtubule-based motor, and immune response were identified. Eight pathways were common to all three comparisons (i.e., calcium signaling, Ras-associated protein 1 signaling, ubiquitin-mediated proteolysis, vascular smooth muscle contraction, oxytocin signaling, and pathway in cancer). The current findings support the role of intracellular ion imbalance, particularly Ca2+, oxidative stress, and impaired programmed cell death on WS progression.

Mechanical and antibacterial properties of the chitosan coated cellulose paper for packaging applications: Effects of molecular weight types and concentrations of chitosan.

Chitosan with low (25 kDa) and high molecular weight (2100 kDa) were used to enhance performances of paper made from steam-exploded bamboo fibers and nanofibrillated cellulose. Chitosan solutions with concentrations of 0-1.0 wt% were manually applied on paper surface using a facile coating approach with a wire bar. Effects of chitosan coatings on morphology, thermal stability, wettability, mechanical performances and antibacterial properties of the paper were investigated. The larger improvement in the mechanical properties and wettability of the chitosan coated paper was observed with increasing concentrations of chitosan due to the disappearance of empty pores between fibers within a cellulose network by the formed chitosan matrix. These improvements were significantly higher when high molecular weight chitosan was applied. Yet, the addition of chitosan slightly decreased the thermal stability of the coated paper, and the chitosan coating did not improve the antimicrobial properties of the paper. The antimicrobial activity of chitosan against Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli) was found to be diminished when a chitosan solution was entrapped within paper. Together with the overall migration of the paper in food simulants, the results suggested that the chitosan coated paper could be applied for non-food-direct-contact packaging materials.