The matricellular protein CCN5 prevents anti-VEGF drug-induced epithelial-mesenchymal transition of retinal pigment epithelium.

Age-related macular degeneration (AMD) is one of the major causes of blindness in the elderly worldwide. Anti-vascular endothelial growth factor (VEGF) drugs have been widely used to treat the neovascular type of AMD (nAMD). However, VEGF acts not only as a pro-angiogenic factor but also as an anti-apoptotic factor in the eyes. In this study, we found that anti-VEGF drugs, including bevacizumab (Bev), ranibizumab (Ran), and aflibercept (Afl), induced epithelial-mesenchymal transition (EMT) in ARPE-19 cells in vitro, accompanied by the induction of CCN2, a potent pro-fibrotic factor. Similarly, intravitreal injection of Afl into mouse eyes resulted in EMT in the retinal pigmented epithelium (RPE). Co-treatment with CCN5, an anti-fibrotic factor that down-regulates CCN2 expression, significantly attenuated the adverse effects of the anti-VEGF drugs both in vitro and in vivo. Inhibition of the VEGF signaling pathway with antagonists of VEGF receptors, SU5416 and ZM323881, induced EMT and up-regulated CCN2 in ARPE-19 cells. Additionally, knock-down of CCN2 with siRNA abolished the adverse effects of the anti-VEGF drugs in ARPE-19 cells. Collectively, these results suggest that anti-VEGF drugs induce EMT in RPE through the induction of CCN2 and that co-treatment with CCN5 attenuates the adverse effects of anti-VEGF drugs in mouse eyes.

Modified mRNA-Mediated CCN5 Gene Transfer Ameliorates Cardiac Dysfunction and Fibrosis without Adverse Structural Remodeling.

Modified mRNAs (modRNAs) are an emerging delivery method for gene therapy. The success of modRNA-based COVID-19 vaccines has demonstrated that modRNA is a safe and effective therapeutic tool. Moreover, modRNA has the potential to treat various human diseases, including cardiac dysfunction. Acute myocardial infarction (MI) is a major cardiac disorder that currently lacks curative treatment options, and MI is commonly accompanied by fibrosis and impaired cardiac function. Our group previously demonstrated that the matricellular protein CCN5 inhibits cardiac fibrosis (CF) and mitigates cardiac dysfunction. However, it remains unclear whether early intervention of CF under stress conditions is beneficial or more detrimental due to potential adverse effects such as left ventricular (LV) rupture. We hypothesized that CCN5 would alleviate the adverse effects of myocardial infarction (MI) through its anti-fibrotic properties under stress conditions. To induce the rapid expression of CCN5, ModRNA-CCN5 was synthesized and administrated directly into the myocardium in a mouse MI model. To evaluate CCN5 activity, we established two independent experimental schemes: (1) preventive intervention and (2) therapeutic intervention. Functional analyses, including echocardiography and magnetic resonance imaging (MRI), along with molecular assays, demonstrated that modRNA-mediated CCN5 gene transfer significantly attenuated cardiac fibrosis and improved cardiac function in both preventive and therapeutic models, without causing left ventricular rupture or any adverse cardiac remodeling. In conclusion, early intervention in CF by ModRNA-CCN5 gene transfer is an efficient and safe therapeutic modality for treating MI-induced heart failure.

Urinary Metabolomic Differentiation of Infants Fed on Human Breastmilk and Formulated Milk.

Human breastmilk is an invaluable nutritional and pharmacological resource with a highly diverse metabolite profile, which can directly affect the metabolism of infants. Application of metabolomics can discriminate the complex relationship between such nutrients and infant health. As the most common biological fluid in metabolomic study, infant urinary metabolomics may provide the physiological impacts of different nutritional resources, namely human breastmilk and formulated milk. In this study, we aimed to identify possible differences in the urine metabolome of 30 infants (1-14 days after birth) fed with breast milk (n = 15) or formulated milk (n = 15). From metabolomic analysis with gas chromatography-mass spectrometry, 163 metabolites from single mass spectrometry (GC-MS), and 383 metabolites from tandem mass spectrometry (GC-MS/MS) were confirmed in urinary samples. Various multivariate statistical analysis were performed to discriminate the differences originating from physiological/nutritional variables, including human breastmilk/formulate milk feeding, sex, and duration of feeding. Both unsupervised and supervised discriminant analyses indicated that feeding resources (human breastmilk/formulated milk) gave marginal but significant differences in urinary metabolomes, while other factors (sex, duration of feeding) did not show notable discrimination between groups. According to the biomarker analyses, several organic acid and amino acids showed statistically significant differences between different feeding resources, such as 2-hydroxyhippurate.

Quantitative Profiling of Carotenoids, Tocopherols, Phytosterols, and Fatty Acids in the Flower Petals of Ten Marigold (Tagetes spp. L.) Cultivars.

Marigold (Tagetes spp.) flower petals are the most vital sources of carotenoids, especially lutein esters, for the production of natural lutein to use for food, feed, and pharmaceutical industries. Several marigold cultivars are cultivated globally; however, their lutein ester composition and contents have not been widely investigated. Considering this, this study aimed to identify and quantify prominent carotenoid esters from the flower petals of ten marigold cultivars by liquid chromatography (LC)-diode-array detection (DAD)-mass spectrometry (MS). In addition, tocopherols, phytosterols, and fatty acids were analyzed by gas chromatography (GC)-flame ionization detection (FID) and GC-MS. Furthermore, the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS•+) and 2,2-diphenyl-1-picrylhydrazyl (DPPH•) radical scavenging abilities of lipophilic extracts were determined. The total carotenoid contents varied significantly (p < 0. 05, Tukey HSD) among cultivars, ranging from 25.62 (cv. Alaska)-2723.11 µg/g fresh weight (cv. Superboy Orange). Among the five major lutein-diesters, (all-E)-lutein-3-O-myristate-3'-O-palmitate and lutein dipalmitate were predominant. Among the studied cultivars, α-tocopherol was recorded, ranging from 167.91 (cv. Superboy Yellow) to 338.50 µg/g FW (cv. Taishan Orange). Among phytosterols, ß-sitosterol was the most prevalent phytosterol, ranging between 127.08 (cv. Superboy Yellow) and 191.99 µg/g FW (cv. Taishan Yellow). Palmitic acid (C16:0; 33.36-47.43%) was the most dominant among the fatty acids. In this study, the highest contents of lutein were recorded from cv. Superboy Orange; however, due to the substantially higher flower petal yield, the cv. Durango Red can produce the highest lutein yield of 94.45 kg/ha. These observations suggest that cv. Durango Red and cv. Superboy Orange are the ideal candidates for lutein fortification in foods and also for commercial lutein extraction.

Dissipation Kinetics and Risk Assessment of Diniconazole, Dinotefuran, Metconazole, and Tebuconazole in Raphanus sativus L.

This study investigated the degradation characteristics and conducted a risk assessment of four pesticides (Diniconazole, Dinotefuran, Metconazole, and Tebuconazole) in the leaves and roots of radish. Radish was cultivated in two greenhouse fields, and samples were collected at 0, 1, 2, 3, 5, 7, and 10 days after pesticide application. Sample analysis was performed using LC-MS/MS, and the recovery rates ranged from 70.1% to 118.6%. The biological half-life of Diniconazole was found to be 6.2 days (leaf and root), Dinotefuran was 5.3 days (leaf) and 4.6 days (root), Metconazole was 9.3 days (leaf) and 3.2 days (root), and Tebuconazole was 8.0 days (leaf) and 5.1 days (root). After comparing the maximum residue limits (MRL) of each pesticide in Korea with the residues during the pre-harvest interval (PHI), Diniconazole showed a Hazard quotient (HQ) exceeding 1, indicating potential risks for true consumers. Furthermore, Tebuconazole showed an HQ of 0.3 or higher, indicating a significant level of risk.

Effects of different stocking density and various phytogenic feed additives dosage levels on growing-finishing pigs.

This study was to investigate the effects of different phytogenic feed additives (PFA) dosage levels in growing- finishing pigs stressed by high stocking density. A total of 72 mix sexed 12 weeks growing pigs ([Landrace × Yorkshire] × Duroc) with initial body weight (BW) of 49.28 ± 4.58 kg were used for 8 weeks. There were 3 replicate pens in each treatment group, with 3 pigs per pen. The dietary treatment groups consisted of basal diets in animal welfare density (negative control [NC]), basal diet in high stocking density (positive control [PC]), PC + 0.04% essential oil (ES1), PC + 0.08% essential oil (ES2), PC + 0.10% bitter citrus extract & essential oil (CES1), PC + 0.20% bitter citrus extract & essential oil (CES2), PC + 0.05% grape pomace extract (GP1), PC + 0.10% grape pomace extract (GP2). The reduction of space allowance decreased (p < 0.05) average daily gain, feed efficiency, and digestibility of dry matter, crude protein, and gross energy. Also, the fecal score of PC groups increased (p < 0.05) compared with other groups. Basic behaviors (feed intake, standing, lying) were inactive (p < 0.05) and singularity behavior (biting) was increased (p < 0.10) under high stocking density. There was no difference in blood profile. However, the supplementation of PFA alleviated the negative effects such as reduced growth performance, nutrient digestibility, and some increasing stress indicators in th blood (cortisol) and animal behavior (biting). In conclusion, the negative effect of high stocking density was most effectively mitigated by the normal dosage of the mixture of bitter citrus extract and essential oil additive (CES1).

Dynamic modeling of pesticide residue in proso millet under multiple application situations.

Proso millet (Panicum miliaceum L.) is a cereal crop with potential resistance to drought and heat stress, making it a promising alternative crop for regions with hot and dry climates. Because of its importance, it is crucial to investigate pesticide residues in proso millet and assess their potential risks to the environment and human health to protect it from insects or pathogens. This study aimed to develop a model for predicting pesticide residues in proso millet using dynamiCROP. The field trials consisted of four plots, with each plot containing three replicates of 10 m2. The applications of pesticides were conducted two or three times for each pesticide. The residual concentrations of the pesticides in the millet grains were quantitatively analyzed using gas and liquid chromatography-tandem mass spectrometry. The dynamiCROP simulation model, which calculates the residual kinetics of pesticides in plant-environment systems, was employed for predicting pesticide residues in proso millet. Crop-specific, environment-specific, and pesticide-specific parameters were utilized to optimize the model. Half-lives of pesticides in grain of proso millet, which were needed to input for dynamiCROP, were estimated using a modified first-order equation. Proso millet-specific parameters were obtained from previous studies. The accuracy of the dynamiCROP model was assessed using statistical criteria, including the coefficient of correlation (R), coefficient of determination (R2), mean absolute error (MAE), relative root mean square error (RRMSE), and root mean square logarithmic error (RMSLE). The model was then validated using additional field trial data, which showed that it could accurately predict pesticide residues in proso millet grain under different environmental conditions. The results demonstrated the accuracy of the model in predicting pesticide residues in proso millet after multiple applications.

Metabolomic Responses of Lettuce (Lactuca sativa) to Allelopathic Benzoquinones from Iris sanguinea Seeds.

Weed management is important in modern crop protection. Chemical weed control using synthetic herbicides, however, suffers from resistance and ecotoxicity. Metabolomic investigation of allelopathy (or allelochemicals) may provide novel alternatives to synthetic herbicides. This study aimed to investigate the detailed metabolomic responses of plants to allelochemicals in Iris seed extracts. The seed extracts of Iris sanguinea showed the strongest growth inhibitory activity against alfalfa, barnyard grass, lettuce, and mustard. 3-Hydroxyirisquinone (3-[10(Z)-heptadecenyl]-2-hydroxy-5-methoxy-1,4-benzoquinone) was isolated as a major allelochemical from I. sanguinea seeds through bioassay-guided fractionation. The compound inhibited the growth of shoots and roots by browning root tips. Discriminant analysis identified 33 differentially regulated lettuce metabolites after treatment with 3-hydroxyirisquinone (3HIQ). Metabolic pathway analysis revealed that several metabolic pathways, including aromatic amino acid biosynthesis and respiratory pathways, were affected by the compounds. Differential responses of membrane lipids (accumulation of unsaturated fatty acids) and extensive formation of reactive oxygen species were observed in root tissues following treatment with 3HIQ. Overall, alkylbenzoquinone from I. sanguinea induced extensive metabolic modulation, oxidative stress, and growth inhibition. The metabolomic responses to allelochemicals may provide fundamental information for the development of allelochemical-based herbicides.

Metabolomic approach of azole fungicides in radish (Raphanus sativus): Perspective of functional metabolites.

Azole fungicides is one of the major fungicides in agricultural field. In this study, toxic effects of diniconazole (DIN), metconazole (MET), and tebuconazole (TEB) to radish leaves and roots were investigated using targeted metabolomics with gas chromatography-mass spectrometry (GC-MS/MS). Especially, the changes of functional chemicals, including phytosterols and glucosinolates evaluated. Radish leaves and roots were harvested after 7 days and 14 days from last exposure. In multivariate analysis, the experimental groups showed clear separation in PCA and PLS-DA score plots. Phytosterols and glucosinolates were significantly changed by azole fungicide. Six metabolic pathways which are affected by fungicides were selected and showed similar patterns regardless of the type of azole fungicide used. As a result, azole fungicide induces the defense mechanisms of plants and affects both primary and secondary metabolism.

Profiling of Nutritionally Vital Bioactive Compounds in Emerging Green Leafy Vegetables: A Comparative Study.

Green leafy vegetables (GLVs), especially lettuce and spinach, are the key source of bioactive antioxidants in a diet. This research compared the contents and composition of lettuce and spinach bioactive compounds with emerging GLVs, moringa and fenugreek. Liquid chromatography (LC)-mass spectrometry (MS) with single ion monitoring (SIM) was used to examine carotenoids and tocols, while phytosterols were examined using gas chromatography (GC)-MS. Among the studied GLVs, the (all-E)-lutein was the most dominating carotenoid ranging between 31.3 (green/red lettuce)−45.3 % (fenugreek) of total carotenoids, followed by (all-E)-violaxanthin and (all-E)-ß-carotene. Surprisingly, (all-E)-ß-carotene, a provitamin A carotenoid, was the second most dominating carotenoid in moringa, accounting for 109.2 µg/g fresh weight (FW). Moreover, the significantly highest (p < 0.05; Tukey HSD) contents of total carotenoids (473.3 µg/g FW), α-tocopherol (83.7 µg/g FW), and total phytosterols (206.4 µg/g FW) were recorded in moringa. Therefore, moringa foliage may serve as an affordable source of nutritionally vital constituents in a diet.

Profiling of Redox-Active Lipophilic Constituents in Leaf Mustard (Brassica juncea (L.) Czern.) Cultivars Using LC-MS and GC-MS.

Leaf mustard is an important commercial and culinary vegetable. However, only limited information is available on the content and composition of the nutritionally important lipophilic constituents in these leaves. This research presents information on the contents and composition of carotenoids, tocols, phytosterols, and fatty acids in four cultivars of leaf mustard. The carotenoids and tocols were analyzed utilizing liquid chromatography (LC)-mass spectrometry (MS) with single ion monitoring (SIM), while phytosterols and fatty acids were analyzed using gas chromatography (GC)-MS and GC-flame ionization detection (FID), respectively. The LC-MS results revealed the dominance of (all-E)-lutein, within the range of 37.12 (cv. Asia Curled)-43.54% (cv. Jeok) of the total carotenoids. The highest amount of all of the individual carotenoids and total carotenoids (143.85 µg/g fresh weight; FW) were recorded in cv. Cheong. Among the studied leaf samples, 67.16 (cv. Asia Curled)-83.42 µg/g FW (cv. Cheong) of α-tocopherol was recorded. Among the phytosterols, ß-sitosterol was the most dominant one among the studied mustard leaves, accounting for 80.42 (cv. Jeok)-83.14% (cv. Red frill) of the total phytosterols. The fatty acid analysis revealed the presence of a significant amount of rare hexadecatrienoic acid (C16:3n3) in the studied mustard leaves, which accounted for 27.17 (cv. Asia Curled)-32.59% (cv. Red frill) of the total fatty acids. Overall, the cv. Cheong represented the highest contents of carotenoids, tocols, and phytosterols. Moreover, cv. Red frill contains the highest amount of n-3 PUFAs and antioxidant compounds. Thus, these cultivars can be promoted in cuisines which can be eaten to obtain the highest health benefits.

Phytogenic feed additives alleviate pathogenic Escherichia coli-induced intestinal damage through improving barrier integrity and inhibiting inflammation in weaned pigs.

BACKGROUND: This study was conducted to investigate the effects of each phytogenic feed additive (PFA; PFA1, bitter citrus extract; PFA2, a microencapsulated blend of thymol and carvacrol; PFA3, a mixture of bitter citrus extract, thymol, and carvacrol; PFA4, a premixture of grape seed, grape marc extract, green tea, and hops; PFA5, fenugreek seed powder) on the growth performance, nutrient digestibility, intestinal morphology, and immune response in weaned pigs infected with Escherichia coli (E. coli). RESULTS: A total of 63 4-week-old weaned pigs were placed in individual metabolic cages and assigned to seven treatment groups. The seven treatments were as follows: 1) NC; basal diet without E. coli challenge, 2) PC; basal diet with E. coli challenge, 3) T1; PC + 0.04% PFA1, 4) T2; PC + 0.01% PFA2, 5) T3; PC + 0.10% PFA3, 6) T4; PC + 0.04% PFA4, 7) T5; PC + 0.10% PFA5. The experiments lasted in 21 d, including 7 d before and 14 d after the first E. coli challenge. In the E. coli challenge treatments, all pigs were orally inoculated by dividing a total of 10 mL of E. coli F18 for 3 consecutive days. The PFA-added groups significantly increased (P < 0.05) average daily gain and feed efficiency and decreased (P < 0.05) the fecal score at d 0 to 14 post-inoculation (PI). Tumor necrosis factor α was significantly lower (P < 0.05) in the PFA-added groups except for T1 in d 14 PI compared to the PC treatment. The T3 had a higher (P < 0.05) immunoglobulin G and immunoglobulin A concentration compared to the PC treatment at d 7 PI. Also, T3 showed significantly higher (P < 0.05) villus height:crypt depth and claudin 1 expression in ileal mucosa, and significantly down-regulated (P < 0.05) the expression of calprotectin compared to the PC treatment. CONCLUSIONS: Supplementation of PFA in weaned pigs challenged with E. coli alleviated the negative effects of E. coli and improved growth performance. Among them, the mixed additive of bitter citrus extract, thymol, and carvacrol showed the most effective results, improving immune response, intestinal morphology, and expression of tight junctions.

Effects of different Bacillus licheniformis and Bacillus subtilis ratios on nutrient digestibility, fecal microflora, and gas emissions of growing pigs.

The objective of this study was to evaluate the effects of different mixing ratios of Bacillus licheniformis and Bacillus subtilis in diets on nutrient digestibility, fecal microflora, and odor gas emissions of growing pigs. A total of four crossbred ([Landrace × Yorkshire] × Duroc) barrows with average body weight (BW) of 41.2 ± 0.7 kg were randomly allotted four diets over four periods in a 4 × 4 Latin square design. Treatments were as follows: Control (CON, basal diet), CON + 0.2% probiotic complex (L4S6, B. licheniformis and B. subtilis at a 4:6 ratio), CON + 0.2% probiotic complex (L5S5, B. licheniformis and B. subtilis at a 5:5 ratio), CON + 0.2% probiotic complex (L6S4, B. licheniformis and B. subtilis at a 6:4 ratio). Dietary probiotic supplementation showed higher crude protein (CP) digestibility values and lower Escherichia coli counts in fecal samples than the CON group (p < 0.05). There was no significant difference in NH3 or H2S emission until day 3. The positive effect of H2S and NH3 emissions was detected earlier with the L4S6 and L5S5 compared to the L6S4, which had a lower ratio of B. subtilis. Both the L4S6 and L5S5 probiotic complexes significantly decreased the fecal H2S and NH3 emission in days 4 and 6 (p < 0.05). On day 7, all probiotic complexes decreased (p < 0.05) H2S and NH3 emissions than the CON group. Our results agreed that the dietary supplementation of Bacillus licheniformis and Bacillus subtilis complexes in growing pigs can significantly improve CP digestibility and reduce fecal E. coli counts, NH3 and H2S emissions. Notably, the higher mixing ratio of Bacillus subtilis in probiotic supplementation is more effective in reducing the odor of manure.

Matricellular Protein CCN5 Gene Transfer Ameliorates Cardiac and Skeletal Dysfunction in mdx/utrn (±) Haploinsufficient Mice by Reducing Fibrosis and Upregulating Utrophin Expression.

Duchenne muscular dystrophy (DMD) is a genetic disorder characterized by progressive muscle degeneration due to dystrophin gene mutations. Patients with DMD initially experience muscle weakness in their limbs during adolescence. With age, patients develop fatal respiratory and cardiac dysfunctions. During the later stages of the disease, severe cardiac fibrosis occurs, compromising cardiac function. Previously, our research showed that the matricellular protein CCN5 has antifibrotic properties. Therefore, we hypothesized that CCN5 gene transfer would ameliorate cardiac fibrosis and thus improve cardiac function in DMD-induced cardiomyopathy. We utilized mdx/utrn (±) haploinsufficient mice that recapitulated the DMD-disease phenotypes and used an adeno-associated virus serotype-9 viral vector for CCN5 gene transfer. We evaluated the onset of cardiac dysfunction using echocardiography and determined the experimental starting point in 13-month-old mice. Two months after CCN5 gene transfer, cardiac function was significantly enhanced, and cardiac fibrosis was ameliorated. Additionally, running performance was improved in CCN5 gene-transfected mice. Furthermore, in silico gene profiling analysis identified utrophin as a novel transcriptional target of CCN5. This was supplemented by a utrophin promoter assay and RNA-seq analysis, which confirmed that CCN5 was directly associated with utrophin expression. Our results showed that CCN5 may be a promising therapeutic molecule for DMD-induced cardiac and skeletal dysfunction.

The TSP-1 domain of the matricellular protein CCN5 is essential for its nuclear localization and anti-fibrotic function.

The matricellular protein CCN5 exerts anti-fibrotic activity in hearts partly by inducing reverse trans-differentiation of myofibroblasts (MyoFBs) to fibroblasts (FBs). CCN5 consists of three structural domains: an insulin-like growth factor binding protein (IGFBP), a von Willebrand factor type C (VWC), and a thrombospondin type 1 (TSP-1). In this study, we set out to elucidate the roles of these domains in the context of the reverse trans-differentiation of MyoFBs to FBs. First, human cardiac FBs were trans-differentiated to MyoFBs by treatment with TGF-ß; this was then reversed by treatment with recombinant human CCN5 protein or various recombinant proteins comprising individual or paired CCN5 domains. Subcellular localization of these recombinant proteins was analyzed by immunocytochemistry, cellular fractionation, and western blotting. Anti-fibrotic activity was also evaluated by examining expression of MyoFB-specific markers, α-SMA and fibronectin. Our data show that CCN5 is taken up by FBs and MyoFBs mainly via clathrin-mediated endocytosis, which is essential for the function of CCN5 during the reverse trans-differentiation of MyoFBs. Furthermore, we showed that the TSP-1 domain is essential and sufficient for endocytosis and nuclear localization of CCN5. However, the TSP-1 domain alone is not sufficient for the anti-fibrotic function of CCN5; either the IGFBP or VWC domain is needed in addition to the TSP-1 domain.

Partial Replacement of Animal Fat with Full-Fat Almond in Broiler Chicken Diets: Performance, Nutrient Digestibility, Blood Profile, Cecal-Fecal Microflora Composition, and Foot-Pad Dermatitis.

The purpose of this study was to evaluate the effect of full-fat almonds (FFA) as an alternative protein and fat source for broiler feed on broiler productivity, nutrient digestibility, blood characteristics, cecal-fecal microflora, and foot-pad dermatitis (FPD). A total of 96, one-day-old broiler chickens (Arbor Acres) with initial body weight 41.61 ± 0.36 g were placed in 16 cages. In each trial, four treatments were set up: a basal diet partially replacing animal fat with FFA 0% (Control, CON), a basal diet partially replacing animal fat with FFA 1% (T1), a basal diet partial replacing animal fat with FFA 2% (T2), a basal diet partially replacing animal fat with FFA 4% (T3). The experiment was conducted for a total of 4 weeks. Feed conversion ratio (FCR) was higher (p < 0.05) in the T3 group of broilers at weeks 0 to 1 than in the CON group of broilers. From weeks 3 to 4, and for the entire experimental period, FCR was lower (p < 0.05) in the T3 group of broilers than in the CON and T1 groups of broilers. The apparent ileal digestibility (AID) of the ether extract (EE) was higher (p < 0.05) in the T3 group than in the other treatment groups, and AID of crude protein (CP) was higher (p < 0.05) in the T3 group than in the CON group. The apparent total tract digestibility (ATTD) of EE was lower (p < 0.05) in the CON group than in the other treatment groups, and the ATTD of CP and energy was higher (p < 0.05) in the T3 group of broilers than in the CON group of broilers. The AID and ATTD of total amino acids were higher (p < 0.05) in the T3 group than in the other treatment groups. Blood cholesterol levels were lower (p < 0.05) in the T3 group of broilers than in the CON and T1 groups of broilers, and higher (p < 0.05) in the CON group of broilers than in the T2 and T3 groups of broilers. The amount of E. coli in the cecal and fecal was lower (p < 0.05) in the T3 group than in CON and T1 groups. FPD score was higher (p < 0.05) in the T3 group of broilers than in the CON group of broilers. In conclusion, replacing a partial of animal fat with at least 4% FFA in broiler diets can increase growth performance and nutrient digestibility in broiler nutrition.

Effects of Replacing Medical Zinc Oxide with Different Ratios of Inorganic: Organic Zinc or Reducing Crude Protein Diet with Mixed Feed Additives in Weaned Piglet Diets.

One hundred twenty weaned piglets (9.34 ± 0.74 kg) were used in a four-week experiment to investigate the effects of replacing medical ZnO with a different ratio of inorganic and organic zinc (IZ:OZ) or a low-crude-protein diet (LP) with mixed feed additives (MFAs) in the weaned piglets' diet. The dietary treatments included a control (CON), T1 (T1; ZnO 1000 mg/kg), T2 (IZ:OZ 850:150), T3 (IZ:OZ 700:300), T4 (IZ:OZ, 500:500), and T5 (LP with MFAs (0.1% essential oils + 0.08% protease + 0.02% xylanase)). The growth performance was decreased (p < 0.05) in the CON treatment compared with the T4 treatment. The diarrhea incidence was decreased (p < 0.05) in the T4 and the T5 treatment compared with the CON and the T1 treatments. The apparent total tract digestibility (ATTD) of nutrients were increased (p < 0.05) in the T4 and T5 treatments compared with the CON, T1, and T2 treatments. The T4 treatment had a higher (p < 0.05) ATTD of zinc than the T1, T2, and T3 treatments. The fecal microflora was improved (p < 0.05) in the T5 treatment compared with the CON and T3 treatments. In conclusion, IZ:OZ 500:500 could improve growth performance, nutrient digestibility, and zinc utilization while reducing diarrhea incidence in weaned piglets. Moreover, LP with MFA could replace medical ZnO.

Anticancer Potential of Lipophilic Constituents of Eleven Shellfish Species Commonly Consumed in Korea.

The present study was aimed to investigate the composition and contents and the major lipophilic compounds, including the sterols, fatty acids, and tocols of shellfish species. Moreover, to explore the antitumor activity of these lipophilic constituents, their cytotoxicity potentials were determined against five different human cancer cells, including colon carcinoma (HCT116), epithelial melanoma (A2058), glioblastoma multiforme (T98G), lung carcinoma (A549), and adenocarcinoma (HeLa). The results show a significant variation in the contents and composition of lipophilic constituents among the studied species. The highest omega-3 (n-3) polyunsaturated fatty acids (PUFAs) were recorded from arrow squid and pacific oysters, accounting for 53.2% and 53.0% of their total fatty acids, respectively. However, the highest cholesterol content was also recorded in arrow squid (154.4 mg/100 g; 92.6% of total sterols). In contrast, in the Japanese littleneck, Yesso scallop, and common orient clam, cholesterol was just 17.1%, 18.3%, and 18.9% of total sterols, respectively, making them the richest source of non-cholesterol sterols (NCS). Lipids extracted from shellfish species showed ABTS+•- and DPPH•-scavenging activities. In the cytotoxicity analysis, lipids extracted from the Argentine red shrimp showed the highest cytotoxicity against glioblastoma multiforme T98G cells, with an IC50 value of 12.3 µg/mL. The composition and cytotoxicity data reported herein may help explore the nutritional and anticancer potentials of shellfish species.

Phytosterol Profiling of Apiaceae Family Seeds Spices Using GC-MS.

Phytosterols are nutritionally vital phytoconstituent owing to their cholesterol (low-density plasma lipoprotein-cholesterol, LDL-C)-lowering, anti-inflammatory, and antioxidant properties. Among the widely used spices and herbs, the seeds spices of the Apiaceae family represented the healthiest fatty acid profile. Thus, to explore the other health-beneficial lipids, the present study was aimed to analyze the phytosterol profile of eight seed spices of the Apiaceae family, utilizing gas chromatography (GC)-mass spectrometry (MS). The sterols contents calculated on an oil (mg/100 g of oil) and spice weight (mg/100 g spices; dry weight) basis varied significantly among the seed spices (p < 0.05; Turkey HSD). The ß-sitosterol and stigmasterol were the most dominating sterols among the studied spices, together accounted for 40.3 (Ajwain) to 69.8% (celery) of total sterols in the seed oil. Among the studied spices, the oil extracted from caraway seeds showed the highest total sterols (602.2 mg/100 g of oil). Interestingly, based on spice weight, fennel seeds also showed the similar highest number of total sterols (134.2 mg/100 g in fennel and 133.3 mg/100 g in caraway), owing to the high contents of oil (25.9%) in fennel seeds. Overall, celery, caraway, fennel, and anise seeds oil are rich sources of health-beneficial phytosterols.

Evaluation of pig behavior changes related to temperature, relative humidity, volatile organic compounds, and illuminance.

The objective of this study was evaluation of pig behavior changes related to temperature, relative humidity, volatile organic compounds (VOCs), and illuminance. A total of 24 growing pigs ([Yorkshire × Landrace] × Duroc) were used in the experiment. A sensor was installed at a height of 0.5 m in the center of the pig house. In experiment 1, temperature was changed every four days to 18°C (T1), 22°C (T2), 26°C (T3), and then 30°C (T4). In experiment 2, relative humidity was adjusted to 45% (low humidity [LH]), 60% (middle humidity [MH]), and then 75% (high humidity [HH]) for four days. In experiment 3, after cleaning the pig house just before experiment, only minimal ventilation was provided. VOCs and pig behaviors were observed for 7 days without cleaning the pig house. In experiment 4, three light bulbs of 40 W (470 lumens / 45 lx; low illuminance [LI]), 75 W (1,055 lumens / 103 lx; middle illuminance [MI]), and 100 W (1,521 lumens / 146 lx; high illuminance [HI]) were used for four days each. Pig behavior analysis was performed for following criteria : Feed intake, Standing, Lying, Sitting, Drink water, Rooting, Posture transition (lying-standing), Posture transition (standing-lying), Wallowing, and Biting. In experiment 1, feed intake time was lower (p < 0.05) for the T3 than other treatment groups. Standing time was highest (p < 0.05) for the T1 and lowest (p < 0.05) for the T3. Lying time was shorter (p < 0.05) in T1 and T2 compared to T3 and T4. Drinking frequency was higher (p < 0.05) for the T4 than other treatment groups. In experiment 2, the frequency of rooting and wallowing increased (p < 0.05) with increasing humidity. LH showed the lowest (p < 0.05) rooting frequency and HH showed the highest (p < 0.05) rooting frequency. In experiment 3, VOCs concentration did not (p > 0.05) change pig behavior. In experiment 4, lying time was the longest (p < 0.05) at LI and shortest (p < 0.05) at HI. Therefore, pig behavior is heavily influenced by the environment, especially temperature and humidity. However, correlation between pig behavior to VOCs and illuminance seems to be needed more research.