Transcriptomic analysis of the longissimus thoracis muscle in pigs has identified molecular regulatory patterns associated with meat quality.

Meat quality is a critical aspect of pig breeding. In addition to genetics, meat quality is also influenced by nutritional and environmental factors. In this study, three pig breeds, Shengxianhua, Jiaxing, and Qinglian Black (SXH, JXB and QLB), were used as experimental animals. Transcriptional analysis was performed on the longissimus thoracis (LT) muscle to investigate variations in intramuscular fat (IMF), inosine monophosphate (IMP), amino acids, and muscle fiber morphology across different breeds. Ingenuity canonical pathway analysis (IPA) identified biological processes and key driver genes related to metabolism and muscle development. Additionally, weighted gene co-expression network analysis (WGCNA) revealed gene modules associated with IMP. KEGG and GO analyses identified specific biological processes and signaling pathways related to IMP, including the Oxidative Phosphorylation pathway and rRNA Metabolic Processes. These findings provide novel insights into the molecular regulatory mechanisms underlying meat quality variations among pig breeds.

CBP22, a Novel Bacteriocin Isolated from Clostridium butyricum ZJU-F1, Protects against LPS-Induced Intestinal Injury through Maintaining the Tight Junction Complex.

A novel bacteriocin secreted by Clostridium butyricum ZJU-F1 was isolated using ammonium sulfate fractionation, cation exchange chromatography, affinity chromatography, and reverse-phase high-performance liquid chromatography (RP-HPLC). The bacteriocin, named CBP22, contained 22 amino acids with the sequence PSAWQITKCAGSIAWALGSGIF. Analysis of its structure and physicochemical properties indicated that CBP22 had a molecular weight of 2264.63 Da and a +1 net charge. CBP22 showed activity against E. col K88, E. coli ATCC25922, and S. aureus ATCC26923. The effects and potential mechanisms of bacteriocin CBP22 on the innate immune response were investigated with a lipopolysaccharide- (LPS-) induced mouse model. The results showed that pretreatment with CBP22 prevented LPS-induced impairment in epithelial tissues and significantly reduced serum levels of IgG, IgA, IgM, TNF-α, and sIgA. Moreover, CBP22 treatment increased the expression of the zonula occludens and reduced permeability as well as apoptosis in the jejunum in LPS-treated mice. In summary, CBP22 inhibits the intestinal injury and prevents the gut barrier dysfunction induced by LPS, suggesting the potential use of CBP22 for treating intestinal damage.

Degradation of sulfanilamide by Fenton-like reaction and optimization using response surface methodology.

Excess sulfonamides are discharged into the environmental system due to the abuse of antibiotics, which threatens the ecological environment and human health. In this study, the ferric and ferrous as well as calcium peroxide (CP), sodium percarbonate (SPC) and sodium persulfate (SPS) have been used to build Fenton-like system for the sulfanilamide (SA) removal. Compared with other Fenton-like system, the Fe3+/CP system exhibited better degradation capacity and 94.65% SA was removed with 3.0 mM CP and 3.0 mM Fe3+. A response surface and corresponding quadratic regression equation were obtained by using a three-level Box-Behnken factorial design with the initial pH value and the dosage of Fe3+ and CP as the model parameters. Depended on the result of the response surface, the optimum conditions of the removal of SA in Fe3+/CP system could be obtained: [Fe3+] = 2.96 mM, [CaO2] = 2.33 mM and [pH] = 6.45. Besides that, the influences of Na+, Mg2+, Cl-, HCO3-, NO3- and HA on SA removal were also investigated under the optimum condition. The results revealed that the high concentration of HCO3- was able to inhibit degradation of SA while other ions and HA have little effect on SA degradation. These results provided a novel strategy to evaluate the catalyst/oxidant system by combining experiment and computer simulation in wastewater treatment.

Integrated rumen microbiome and serum metabolome analysis responses to feed type that contribution to meat quality in lambs.

BACKGROUND: Lifestyle factors, such as diet, are known to be a driver on the meat quality, rumen microbiome and serum metabolites. Rumen microbiome metabolites may be important for host health, the correlation between rumen microbiome and production of rumen metabolites are reported, while the impact of rumen microbiome on the serum metabolome and fatty acid of meat are still unclear. This study was designed to explore the rumen microbiome, serum metabolome and fatty acid of meat in response to the grass diet and concentrate diet to lambs, and the relationship of which also investigated. METHODS: In the present study, 12 lambs were randomly divided into two groups: a grass diet (G) and a concentrate diet (C). Here, multiple physicochemical analyses combined with 16S rRNA gene sequences and metabolome analysis was performed to reveal the changes that in response to feed types. RESULTS: The concentrate diet could improve the growth performance of lambs compared to that fed with the grass diet. The microbiome composition was highly individual, compared to the concentrate group, the abundance of Rikenellaceae_RC9_gut_group, F082_unclassified, Muribaculaceae_unclassified, Ruminococcaceae_NK4A214_group, Bacteroidetes_unclassified, and Bacteroidales_UCG-001_unclassified were significantly (P < 0.05) lower in the grass group, while, the abundance of Succinivibrio, Succinivibrionaceae_UCG-002, Fibrobacter and Christensenellaceae_R-7_group were significantly (P < 0.05) higher in the grass group. Serum metabolomics analysis combined with enrichment analysis revealed that serum metabolites were influenced by feed type as well as the metabolic pathway, and significantly affected serum metabolites involved in amino acids, peptides, and analogues, bile acids, alcohols and derivatives, linoleic acids derivatives, fatty acids and conjugates. Most of the amino acids, peptides, and analogues metabolites were positively associated with the fatty acid contents. Among the bile acids, alcohols and derivatives metabolites, glycocholic was positively associated with all fatty acid contents, except C18:0, while 25-Hydroxycholesterol and lithocholic acid metabolites were negatively associated with most of the fatty acid contents. CONCLUSION: Correlation analysis of the association of microbiome with metabolite features, metabolite features with fatty acid provides us with comprehensive understanding of the composition and function of microbial communities. Associations between utilization or production were widely identified among affected microbiome, metabolites and fatty acid, and these findings will contribute to the direction of future research in lamb.

Endogenous chitinase might lead to differences in growth performance and intestinal health of piglets fed different levels of black soldier fly larva meal.

This study aimed to investigate the effects of different levels of black soldier fly (BSF) replacing soybean meal (SBM) in diets on the performance and health condition of piglets. A total of 180 weaned piglets were allocated into 5 treatments: BSF0 (corn-soybean meal basal diet), BSF25 (BSF replacing 25% SBM), BSF50 (BSF replacing 50% SBM), BSF75 (BSF replacing 75% SBM) and BSF100 (BSF replacing 100% SBM). During the whole period, in comparison with BSF0, average daily gain (ADG) and average daily feed intake increased in the BSF25 and BSF50 groups, whereas ADG decreased in the BSF75 and BSF100 groups (P < 0.05). The result of quadratic fitting curve showed that piglets exhibited the highest ADG when BSF replaced around 20% SBM. Compared with BSF0, organic matter and dry matter digestibility improved in the BSF25 group, whereas ether extract digestibility decreased in the BSF100 group (P < 0.05). In comparison with BSF0, piglets from the BSF25 group showed a higher duodenal ratio of villus height to crypt depth, increased jejunal sucrase activity, serum neuropeptide Y and ghrelin levels, elevated ileal immunoglobulin (Ig) A, IgG and IgM contents and a lower leptin level, and piglets from the BSF100 group exhibited an increased relative weight of kidney (P < 0.05). However, no significant differences were observed in the expression level of tight junction proteins and chitin-degrading enzyme. Additionally, compared with BSF0, the abundance of short chain fatty acid producing bacteria such as Ruminococcaceae, Faecalibacterium and Butyricicoccus increased, and potential pathogenic bacteria decreased in piglets from the BSF25 group, whereas piglets from the BSF100 group had a greater abundance of harmful bacteria. In conclusion, BSF replacing 25% SBM in diets could improve digestive parameters, immune function and intestinal microbiota, and thus improved growth performance of piglets. However, BSF replacing 100% SBM showed an adverse effect on piglet performance, and the reason might be related to the limited amount of chitin-degrading enzyme.

Fermented mixed feed alters growth performance, carcass traits, meat quality and muscle fatty acid and amino acid profiles in finishing pigs.

This study was conducted to investigate the effects of fermented mixed feed (FMF) on growth performance, carcass traits, meat quality, muscle amino acid and fatty acid composition and mRNA expression levels of genes related to lipid metabolism in finishing pigs. In the present study, 144 finishing pigs (Duroc × Berkshire × Jiaxing Black) were randomly allocated to 3 dietary treatments with 4 replicate pens per group and 12 pigs per pen. The dietary treatments included a basal diet (CON), a basal diet + 5% FMF and a basal diet + 10% FMF. The experiment lasted 38 d after 4 d of acclimation. The results showed that 5% and 10% FMF significantly increased the average daily gain (ADG) of the females but not the males (P < 0.05), but FMF supplementation showed no impact on carcass traits. Moreover, 10% FMF supplementation increased the meat color45 min and meat color24 h values, while it decreased the shear force relative to CON (P < 0.05). In addition, 10% FMF significantly increased the contents of flavor amino acids (FAA), total essential AA (EAA), total non-EAA (NEAA) and total AA relative to CON (P < 0.05). Furthermore, the diet supplemented with 10% FMF significantly increased the concentration of n-3 polyunsaturated fatty acids (PUFA), n-6 PUFA and total PUFA, and the PUFA to saturated fatty acids ratio (P < 0.05), suggesting that FMF supplementation increased meat quality. Moreover, compared with the CON, 10% FMF supplementation increased the mRNA expression of lipogenic genes, including CEBPα, PPARγ, SREBP1 and FABP4, and upregulated the expression of unsaturated fatty acid synthesis (ACAA1 and FADS2). Together, our results suggest that 10% FMF dietary supplementation improved the female pigs' growth performance, improved the meat quality and altered the profiles of muscle fatty acids and amino acids in finishing pigs. This study provides a reference for the production of high-quality pork.

Fermented mixed feed regulates intestinal microbial community and metabolism and alters pork flavor and umami.

This study aimed to determine the effects of fermented mixed feed (FMF) supplementation (0%, 5% and 10%) on the intestinal microbial community and metabolism, and the compositions of volatile flavor compounds and inosine monophosphate (IMP) contents in the longissimus thoracis. In this study, 144 finishing pigs (Duroc × Berkshire × Jiaxing Black) were randomly allocated to 3 groups with 4 replicate pens per group and 12 pigs per pen. The experiment lasted 38 days after 4 days of acclimation. The 16S rRNA gene sequences and an untargeted metabolomics analysis showed FMF altered the profiles of microbes and metabolites in the colon. Heracles flash GC e-nose analysis showed that 10% FMF (treatment 3) had a greater influence on the compositions of volatile flavor compounds than 5% FMF (treatment 2). Compared to 0% FMF (treatment 1), the contents of total aldehydes, (E,E)-2,4-nonadienal, dodecanal, nonanal and 2-decenal were significantly increased by treatment 3, and treatment 3 increased IMP concentrations and gene expressions related to its synthesis. Correlations analysis showed significantly different microbes and metabolites had strong correlations with the contents of IMP and volatile flavor compounds. In conclusion, treatment 3 regulated intestinal microbial community and metabolism, that in turn altered the compositions of volatile compounds, which contributed to improving pork flavor and umami.

Effects of dietary oil sources and fat extraction methods on apparent and standardized ileal digestibility of fat and fatty acids in growing pigs.

BACKGROUND: There is a lack of data for the standardized ileal digestibility (SID) of fat and fatty acids in national feed databases. In addition, it is important to specify the procedures used for fat analyses. Therefore, an experiment was conducted to 1) determine the apparent ileal digestibility (AID) and SID of fat and fatty acids in ten different oil sources for growing pigs and to develop prediction equations for SID of fat based on fatty acid composition; and 2) compare the effect of the fat extraction methods on the calculated values for endogenous loss and digestibility of fat. METHODS: Twenty-two barrows (initial body weight: 32.1 ± 2.3 kg) were surgically fitted with a T-cannula in the distal ileum, and allotted to 1 of 11 experimental diets in a 4-period Youden Square design. A fat-free diet was formulated using cornstarch, soy protein isolate and sucrose. Ten oil-added diets were formulated by adding 6% of dietary oil sources to the fat-free diet at the expense of cornstarch. All diets contained 26% sugar beet pulp and 0.40% chromic oxide. RESULTS: The endogenous loss of ether extract (EE) was lower than that of acid-hydrolyzed fat (AEE; P < 0.01). There were significant differences in the AID and SID of fat and saturated fatty acids across the dietary oil sources (P < 0.05). The SID of AEE for palm oil was lower than that of sunflower oil, corn oil, canola oil, rice oil and flaxseed oil (P < 0.01). The AID and SID of fat ranged from 79.65% to 86.97% and from 91.14% to 99.18%. Although the AID of EE was greater than that of AEE (P < 0.01), there was no significant difference in SID of EE and AEE except for palm oil. The ratio of unsaturated to saturated fatty acids (U/S) had a positive correlation with SID of fat (P < 0.05), whereas C16:0 and long chain saturated fatty acids (LSFA) were significant negatively correlated with SID of fat (P < 0.01). The best-fit equation to predict SID of fat was SID AEE = 102.75 - 0.15 × LSFA - 0.74 × C18:0 - 0.03 × C18:1 (Adjusted coefficient of determination = 0.88, P < 0.01). CONCLUSIONS: When calculating the SID of fat, the EE content of the samples can be analyzed using the direct extraction method, whereas the acid hydrolysis procedure should be used to determine the AID of fat. Fat digestibility of dietary oils was affected by their fatty acid composition, especially by the contents of C16:0, LSFA and U/S.

Protective effects of polysaccharides from Atractylodes macrocephalae Koidz. against dextran sulfate sodium induced intestinal mucosal injury on mice.

In the present research, the water-soluble polysaccharides (AMP) from Atractylodes macrocephalae Koidz. were isolated and prepared. The protective effects of AMP on intestinal mucosal barrier injury induced by dextran sulfate sodium (DSS) in mice were investigated. It was found that AMP treatment significantly alleviated the body weight decreases and shorten colon length, and ameliorated colonic damage induced by DSS. Importantly, AMP prevented the over-expression of proinflammatory cytokines TNF-α, IL-1ß and IL-6, and decreased the infiltration of neutrophils in colon. Additionally, AMP could raise expressions of Mucin 2 and tight junction protein Claudin-1. AMP also modulated the intestinal microbiota by enhancing the overall richness and diversity, greatly reducing the proportion of harmful bacteria, for instance, Clostridiumsensu stricto1 and Escherichia Shigella, however, augmenting the ratio of potential beneficial bacteria such as Faecalibaculum and Bifidobacterium. This work offers some important insights on protective effects of polysaccharides AMP against intestinal barrier dysfunction and provides underlying mechanism of health-beneficial properties of these biological macromolecules.

Clostridium Butyricum ZJU-F1 Benefits the Intestinal Barrier Function and Immune Response Associated with Its Modulation of Gut Microbiota in Weaned Piglets.

This study investigated the effects of dietary C. butyricum ZJU-F1 on the apparent digestibility of nutrients, intestinal barrier function, immune response, and microflora of weaned piglets, with the aim of providing a theoretical basis for the application of Clostridium butyricum as an alternative to antibiotics in weaned piglets. A total of 120 weanling piglets were randomly divided into four treatment groups, in which piglets were fed a basal diet supplemented with antibiotics (CON), Bacillus licheniformis (BL), Clostridium butyricum ZJU-F1 (CB), or Clostridium butyricum and Bacillus licheniformis (CB-BL), respectively. The results showed that CB and CB-BL treatment increased the intestinal digestibility of nutrients, decreased intestinal permeability, and increased intestinal tight junction protein and mucin expression, thus maintaining the integrity of the intestinal epithelial barrier. CB and CB-BL, as exogenous probiotics, were also found to stimulate the immune response of weaned piglets and improve the expression of antimicrobial peptides in the ileum. In addition, dietary CB and CB-BL increased the proportion of Lactobacillus. The levels of butyric acid, propionic acid, acetic acid, and total acid were significantly increased in the ceca of piglets fed CB and CB-BL. Furthermore, we validated the effects of C. butyricum ZJU-F1 on the intestinal barrier function and immune response in vitro and found C. butyricum ZJU-F1 improved intestinal function and enhanced the TLR-2-MyD88-NF-κB signaling.

Dramatic enhancement effects of l-cysteine on the degradation of sulfadiazine in Fe3+/CaO2 system.

Excessive sulfonamides accumulated in soil and groundwater seriously menace the ecological environment and human health. The performance of a Fenton-like system applying Fe3+ and calcium peroxide (CaO2) in the presence of l-cysteine(l-cys) for sulfadiazine (SDZ) degradation was investigated. Compared with other chelating agents such as citric acid, butyric acid and Ethylenediaminetetraacetic acid, l-cys could effectively promote the SDZ removal in Fe3+/CaO2 system. With the addition of 0.5 mM l-cys, the SDZ degradation increased from 2.14% to 66.53% in 60 min. High concentration of HCO3- inhibited the degradation of SDZ while slightly negative effects on SDZ degradation were observed in the presence of Cl- or humic acid (HA) in l-cys/Fe3+/CaO2 system. Electron paramagnetic resonance (EPR) analysis and radicals scavenge tests affirmed the generation of OH and O2- in l-cys/Fe3+/CaO2 system. Possible degradation pathway of SDZ was speculated and the toxicity of SDZ intermediates was further evaluated. l-cys could enhance the reduction of Fe3+ to Fe2+ and reduced the Fe3+ precipitation due to the l-cys could form stable complexes with Fe3+. l-cys/Fe3+/CaO2 system exhibited high mineralization ability. Overall, these results indicated that l-cys is a promising chelating agent for sulfadiazine wastewater treatment.

Quantitative study of taste bud distribution within the oral cavity of the postnatal mouse.

The aim of this study was to investigate the age-related developmental changes of taste bud distribution within the subpopulations at different postnatal ages in the mouse oral cavity. Developmental changes of taste bud distribution on the soft palate, fungiform, foliate and circumvallate papillae in the mouse oral cavity were examined histologically at different postnatal ages. After paraffin embedding, complete serial sections at 10mum thickness were made and stained by routine hematoxylin-eosin staining methods. Digitised images for each section were examined carefully. The existence of a taste pore was used to identify mature taste buds. A two-way analysis of variance (group versus age) was used to analyse differences in taste bud number and characteristics for each of the developmental changes. An independent measures t-test was used to compare two means. No taste buds with pores were observed at birth within circumvallate and foliate papillae. However, 61% of the circumvallate and 58% of the foliate taste buds contained taste pores at 2 weeks after birth. In contrast, at birth, 55% of the taste buds on the soft palate and only 22% of the taste buds within fungiform papillae contained taste pores. Then, the number of mature taste buds (taste buds with pores) increased rapidly 1 week after birth, resulting in 90% of soft palate taste buds and 32% of fungiform taste buds containing taste pores. These results suggests that the earlier maturation of soft palate taste buds compared with the other populations in the oral cavity raises evidence of their significant role in the taste mechanism, especially in the early life of the mouse.

LFP-20, a porcine lactoferrin peptide, ameliorates LPS-induced inflammation via the MyD88/NF-κB and MyD88/MAPK signaling pathways.

LFP-20 is one of the 20 amino acid anti-microbial peptides identified in the N terminus of porcine lactoferrin. Apart from its extensively studied direct anti-bacterial activity, its potential as an activator of immune-related cellular functions is unknown. Therefore, this study investigated its anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated pig alveolar macrophages in vitro and systemic inflammation in an in vivo mouse model. We found that the inhibitory effects of LFP-20 on production of pro-inflammatory cytokines were independent of its LPS-binding activity. However, they were associated with NF-κB and MAPK-dependent signaling. Furthermore, LFP-20 might directly influence MyD88 levels to block its interaction with NF-κB and MAPK-dependent signaling molecules that might alter LPS-mediated inflammatory responses in activated macrophages. Taken together, our data indicated that LFP-20 prevents the LPS-induced inflammatory response by inhibiting MyD88/NF-κB and MyD88/MAPK signaling pathways, and sheds light on the potential use of LFP-20 in the therapy of LPS-mediated sepsis.

Antimicrobial peptide Cathelicidin-BF prevents intestinal barrier dysfunction in a mouse model of endotoxemia.

Intestinal barrier functions are altered during the development of sepsis. Cathelicidin antimicrobial peptides, such as LL-37 and mCRAMP, can protect animals against intestinal barrier dysfunction. Cathelicidin-BF (C-BF), a new cathelicidin peptide purified from the venom of the snake Bungarus fasciatus, has been shown to have both antimicrobial and anti-inflammatory properties. This study investigated whether C-BF pretreatment could protect the intestinal barrier against dysfunction in a mouse model of endotoxemia, induced by intraperitoneal injection of LPS (10mg/kg). Mice were treated with low or high dose C-BF before treatment with LPS, and samples were collected 5h after LPS treatment. C-BF reduced LPS induced intestinal histological damage and gut permeability to 4 KD Fluorescein-isothiocyanate-conjugated dextran. Pretreatment with C-BF prevented LPS induced intestinal tight junction disruption and epithelial cell apoptosis. Moreover, C-BF down regulated the expression and secretion of TNF-α, a process involving the NF-κB signaling pathway. C-BF also reduced LPS induced TNF-α expression through the NF-κB signaling pathway in mouse RAW 264.7 macrophages. These findings indicate that C-BF can prevent gut barrier dysfunction induced by LPS, suggesting that C-BF may be used to develop a prophylactic agent for intestinal injury in endotoxemia.

A novel sweet taste cell-based sensor.

In this paper, a novel sweet taste cell-based sensor is proposed for tastants detection. The human colorectal carcinoma NCI-H716 cell lines, which express α-gustducin and sweet taste receptor T1R1/T1R3, are cultured on the carbon screen-printed electrode with the pre-coated poly-L-ornithine and lamimin for adhesion in extracellular matrix. The electrode is placed into a fluidic and environment regulation system. When stimulated by an electric field at a fixed frequency of 1 kHz and 10 µA, the electrochemical impedance spectrum data was recorded by EG&G 273A and high performance dual phase analog lock-in amplifier 5210 and processed by bistable stochastic resonance method. Four basic tastants and sucrose solutions in seven concentrations can be decided by maximums of the signal-to-noise ratio and relevant noise intensity. A negative control experiment utilizing COLO-205 cell lines demonstrate that carbon screen-printed electrode with mammalian cell line without the expression of gust TIR2 and TIR3 lacked the tastant detecting ability. The sensor system presents good stability and repeatability. The proposed sensor is promising for practical applications, and provides a novel way for taste mechanism investigation.

A reliable method to obtain cells of taste buds from fungiform papillae of mice.

Taste buds consist of four kinds of cells which have distinct characteristics and play different roles in recognizing chemical compounds contained in foodstuffs. In this study we describe a procedure for separating viable taste bud cells from the fungiform papillae in mice. After sacrifice with CO(2), the mouse tongue was excised and immediately incubated in collagenase II and dispase II. The epithelium with fungiform papillae was then peeled away from underlying tissue and the anterior one-third region was incubated in a solution of 0.25% trypsin and 0.02M ethylene-diamine-tetraacetic acid (EDTA) for 8-12min. Following incubation, a cell suspension was obtained by mechanical dissociation. Cells in suspension were identified as taste bud cells by their morphology and by immunofluorescence. A 0.25% trypan blue staining demonstrated that nearly 90% of these cells remained viable. Micrographs from scanning electron microscopy illustrated that taste buds were dissociated from the fungiform papillae, while maintaining the integrity of the other part of the dissociated lingual epithelium during incubation. Such a method allows acquisition of viable taste cells and will aid further research in the study of gustatory characteristics.