Effects of enzyme levels in total mixed ration containing oil palm frond silage on intake, rumen fermentation, and growth performance of male goat

This experiment was conducted to study the effects of supplementing the total mixed ration (TMR) containing oil palmfrond (OPF) silage with different levels of enzyme on feed intake and growth performance of goat. Twenty four post-weaningBoer  Thai Native crossbred male goats with initial body weight (BW) of 11-18 kg, were arranged to receive four dietarytreatments in a randomized complete block design. The diet used in the study contained 60% oil palm frond silage and 40%concentrate. The enzyme mixture produced by Aspergillus spp. BCC 274, containing approximately 1107, 9106, 2106, 1106and 2106 unit/kg dry weight for xylanase, -glucanase, cellulase, mannanase and amylase, respectively, was supplementedto the concentrate portion at 0, 2, 4 and 6 g/kgDM of the TMR. The results showed that the supplementation of enzyme tothe TMR did not affect (P>0.05) dry matter intake (DMI). Goats receiving TMR supplemented with enzyme at 2 g/kgDMtended to have higher ADG and better feed per gain ratio as compared with other treatments. Coefficient of DM digestibilityof TMR was not significantly affected by the enzyme supplementation. In addition, there were no significant differences(P>0.05) among treatments regarding, average NH3-N concentration, the amount of C2, C3 and C4 in the rumen fluid and BUNconcentration. However, overall mean of ruminal NH3-N concentration was significantly lower in goat receiving TMR supplemented with enzyme at 2 g/kgDM than that of goat receiving TMR with no enzyme supplementation (P<0.05). Based on thisexperiment, the application of enzyme at 2 g/kgDM in TMR containing OPF silage could increase ruminal availability of slowlydigestible carbohydrate and improve goat performance

Monitoring of Chicken RNA Integrity as a Function of Prolonged Postmortem Duration

Gene expression profiling has offered new insights into postmortem molecular changes associated with meat quality. To acquire reliable transcript quantification, high quality RNA is required. The objective of this study was to analyze integrity of RNA isolated from chicken skeletal muscle (pectoralis major) and its capability of serving as the template in quantitative real-time polymerase chain reaction (qPCR) as a function of postmortem intervals representing the end-points of evisceration, carcass chilling and aging stages in chicken abattoirs. Chicken breast muscle was dissected from the carcasses (n = 6) immediately after evisceration, and one-third of each sample was instantly snap-frozen and labeled as 20 min postmortem. The remaining muscle was stored on ice until the next rounds of sample collection (1.5 h and 6 h postmortem). The delayed postmortem duration did not significantly affect A260/A280 and A260/A230 (p≥0.05), suggesting no altered purity of total RNA. Apart from a slight decrease in the 28s:18s ribosomal RNA ratio in 1.5 h samples (p<0.05), the value was not statistically different between 20 min and 6 h samples (p≥0.05), indicating intact total RNA up to 6 h. Abundance of reference genes encoding beta-actin (ACTB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine-guanine phosphoribosyltransferase (HPRT), peptidylprolylisomerase A (PPIA) and TATA box-binding protein (TBP) as well as meat-quality associated genes (insulin-like growth factor 1 (IGF1), pyruvate dehydrogenase kinase isozyme 4 (PDK4), and peroxisome proliferator-activated receptor delta (PPARD) were investigated using qPCR. Transcript abundances of ACTB, GAPDH, HPRT, and PPIA were significantly different among all postmortem time points (p<0.05). Transcript levels of PDK4 and PPARD were significantly reduced in the 6 h samples (p<0.05). The findings suggest an adverse effect of a prolonged postmortem duration on reliability of transcript quantification in chicken skeletal muscle. For the best RNA quality, chicken skeletal muscle should be immediately collected after evisceration or within 20 min postmortem, and rapidly preserved by deep freezing

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

open9siThis research was financially supported by Cluster and Program Management, National Science and Technology Development Agency (Thailand; project number P15-50668), and by Office of the Permanent Secretary, Ministry of Higher Education, Science, Research and Innovation (Thailand; P20-50946 and P21-50165).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 &lt; 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 &lt; 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 &lt; 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 &lt; 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.openMalila, Yuwares; Uengwetwanit, Tanaporn; Thanatsang, Krittaporn V.; Arayamethakorn, Sopacha; Srimarut, Yanee; Petracci, Massimiliano; Soglia, Francesca; Rungrassamee, Wanilada; Visessanguan, WonnopMalila, Yuwares; Uengwetwanit, Tanaporn; Thanatsang, Krittaporn V.; Arayamethakorn, Sopacha; Srimarut, Yanee; Petracci, Massimiliano; Soglia, Francesca; Rungrassamee, Wanilada; Visessanguan, Wonno

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 64 0.05 and absolute fold change 651.2. 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

Monitoring of white striping and wooden breast cases and impacts on quality of breast meat collected from commercial broilers ()

Objective This study aimed at investigating white striping (WS) and wooden breast (WB) cases in breast meat collected from commercial broilers. Methods A total of 183 breast samples were collected from male Ross 308 broilers slaughtered at the age of 6 weeks (n = 100) and 7 weeks (n = 83). The breasts were subjected to meat defect inspection, meat quality determination and histology evaluation. Results Of 183, 4 breasts from 6-week-old broilers were classified as non-defective while the others exhibited the WS lesion. Among the 6-week-old birds, the defective samples from the medium size birds (carcass weight ≤2.5 kg) showed mild to moderate WS degree with no altered meat quality. Some of the breasts from the 6-week-old birds with carcass weight above 2.5 kg exhibited WB in accompanied with the WS condition. Besides of a reduction of protein content, increases in collagen matter and pH values in the defective samples (p<0.05), no other impaired quality indices were detected within this group. All 7-week-old broilers yielded carcasses weighing above 2.5 kg and showed abnormal characteristics with progressive severity. The breasts affected with severe WS and WB showed the greatest cook loss, hardness, springiness and chewiness (p<0.05). Development of WB induced significantly increased drip loss in the samples (p<0.05). Histology indicated necrotic events in the defective myofibers. Based on logistic regression, increasing percent breast weight by one unit enhanced the chance of WS and WB development with advanced severity by 50.9% and 61.0%, respectively. Delayed slaughter age from 6 to 7 weeks increased the likelihood of obtaining increased WS severity by 56.3%. Conclusion Cases of WS and WB defects in Southeast Asia have been revealed. Despite few cases of the severe WS and WB, such abnormal conditions significantly impaired technological properties and nutritional quality of broiler breasts

Effect of Skipjack Tuna Spleen on the Liquefaction and Characteristics of Sardine Fish Sauce

The effects of the addition of spleen of skipjack tuna (Katsuwonus pelamis), at levels of 0%, 10% and 20%, on the liquefaction and characteristics of fish sauce produced from the sardine (Sardinella gibbosa) with different salt concentrations (15%, 20% and 25%) were monitored during fermentation for 180 days. Fish sauces prepared from sardine with spleen supplementation contained greater total nitrogen, amino nitrogen, formaldehyde nitrogen and ammonia nitrogen than did those without spleen addition throughout the fermentation. The rate of liquefaction was dependent on the amount of spleen added. Reduction of salt content accelerated the hydrolysis of fish protein during fermentation. The liquefaction rate of the lower salt-treated samples was generally faster than were those treated with higher salt content. Among all treatments, sardine with 25% spleen and 15% salt added exhibited the greatest protein hydrolysis, particularly at the early stages, suggesting the combined effects of autolysis and spleen proteinase. The greater liquefaction was coincidental with the development of browning as well as the increase in redness of liquid formed. An acceptability test revealed that the samples were different in colour, aroma, taste and overall acceptance (p < 0.05). Fish sauce samples containing 20% salt, without and with 10% spleen addition had similar acceptabilities to commercial fish sauce. Therefore, the addition of spleen, as well as salt reduction, can accelerate the liquefaction of sardine for fish sauce production

Proteolytic degradation of sardine (Sardinella gibbosa) proteins by trypsin from skipjack tuna (Katsuwonus pelamis) spleen

Trypsin from skipjack tuna (Katsuwonus pelamis) spleen was purified by ammonium sulfate precipitation and a series of chromatographies including Sephacryl S-100 and Benzamidine Sepharose 4 Fast Flow (high sub). The enzyme was purified to 22.3 folds with a yield of 51.6%. The molecular weight of trypsin was estimated to be 42 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Purified trypsin was able to hydrolyze natural actomyosin (NAM) and myosin, but rarely hydrolyzed collagen. Myosin heavy chain was most susceptible to hydrolysis by trypsin as evidenced by the lowest band intensity remained. The effect of NaCl on proteolytic activity was also studied. The band intensity of myosin heavy chain slightly increased as NaCl concentration increased, suggesting the inhibitory activity of NaCl. When hydrolytic activities of skipjack tuna spleen and bovine pancreas trypsins on sardine proteins, including NAM, myosin and collagen were compared, it was found that trypsin from bovine pancreas showed the greater activity towards NAM and myosin than that from skipjack tuna spleen. However, both enzymes could not degrade collagen

Partitioning and recovery of proteinase from tuna spleen by aqueous two-phase systems

Partitioning of spleen proteinase from yellowfin tuna (Thunnus albacores) in an aqueous two-phase system (ATPS) was investigated. Phase compositions including PEG molecular mass and concentration as well as types and concentration of salts affected the protein partitioning. ATPS comprising PEG1000 (15% w/w) and magnesium sulfate (20% w/w) provided the best condition for the maximum partitioning of the proteinase into the top phase and gave a highest specific activity (47.0 units/μg protein) and purification fold (6.61). The yield of 69.0% was obtained. Under the same ATPS condition used, the partitioning of proteinase of splenic extract from three tuna species involving skipjack tuna, yellowfin tuna and tongol tuna were compared. The purity of splenic extract from all tuna species increased after ATPS process. Among all species tested, yellowfin tuna showed the highest purification fold, followed by tongol tuna and skipjack tuna, respectively. SDS-substrate gel electrophoresis revealed that the band intensity of major proteinase in ATPS fraction from all tuna species slightly increased with the concomitant decrease in band intensity of other contaminating proteins, indicating the greater specific activity of splenic extract. Therefore, ATPS was an effective method for partitioning and recovery of proteinases from tuna spleen