Maternal Diet during Pregnancy Alters the Metabolites in Relation to Metabolic and Neurodegenerative Diseases in Young Adult Offspring.

Maternal nutrition during the critical period of pregnancy increases the susceptibility of offspring to the development of diseases later in life. This study aimed to analyze metabolite profiles to investigate the effect of maternal diet during pregnancy on changes in offspring plasma metabolites and to identify correlations with metabolic parameters. Pregnant Sprague-Dawley rats were exposed to under- and overnutrition compared to controls, and their offspring were fed a standard diet after birth. Plasma metabolism was profiled in offspring at 16 weeks of age using liquid chromatography-mass spectrometry (LC-MS/MS) and gas chromatography-tandem mass spectrometry (GC-MS/MS). We analyzed 80 metabolites to identify distinct metabolites and metabolic and neurodegenerative disease-associated metabolites that were sex-differentially altered in each group compared to controls (p < 0.05, VIP score > 1.0). Specifically, changes in 3-indolepropionic acid, anthranilic acid, linoleic acid, and arachidonic acid, which are involved in tryptophan and linoleic acid metabolism, were observed in male offspring and correlated with plasma leptin levels in male offspring. Our results suggest that fatty acids involved in tryptophan and linoleic acid metabolism, which are altered by the maternal diet during pregnancy, may lead to an increased risk of metabolic and neurodegenerative diseases in the early life of male offspring.

Flow Characteristics of the Conjugate of Anti-CD3 Monoclonal Antibodies and Magnetic Nanoparticle in PBS and Blood Vessels.

Previously, anti-CD3 antibodies delivered intravenously have been known for their negative side effects. The experimental conditions for optimal liquid production are derived from the Fc-directed conjugation of anti-CD3 foralumab antibodies and magnetic nanoparticles (Ab-MNPs). The anti-CD3 antibodies are prepared for conjugation with MNPs using SiteClick antibody labelling kits. The successful conjugation of the Ab-MNPs is confirmed using a transmission electron microscopy (TEM) image and an energy dispersive spectroscopy (EDS) analysis. The average values ​​of the moving speed of MNPs and Ab-MNPs in phosphate buffer saline (PBS) were + 3.16 pix/frame and + 6.70 pix/frame in the x-axis, respectively. This implies that MNPs with CD3 antibodies attached to the surface through biocompatible ligand functional groups has better fluidity in PBS. Afterwards, a non-clinical animal testing for the flow characteristics of Ab-MNPs inside a blood vessel is carried out to observe the effects of Ab-MNP delivery through intravenous injection.

Natural Product-Based Potential Therapeutic Interventions of Pulmonary Fibrosis.

Pulmonary fibrosis (PF) is a disease-refractive lung condition with an increased rate of mortality. The potential factors causing PF include viral infections, radiation exposure, and toxic airborne chemicals. Idiopathic PF (IPF) is related to pneumonia affecting the elderly and is characterized by recurring scar formation in the lungs. An impaired wound healing process, defined by the dysregulated aggregation of extracellular matrix components, triggers fibrotic scar formation in the lungs. The potential pathogenesis includes oxidative stress, altered cell signaling, inflammation, etc. Nintedanib and pirfenidone have been approved with a conditional endorsement for the management of IPF. In addition, natural product-based treatment strategies have shown promising results in treating PF. In this study, we reviewed the recently published literature and discussed the potential uses of natural products, classified into three types-isolated active compounds, crude extracts of plants, and traditional medicine, consisting of mixtures of different plant products-in treating PF. These natural products are promising in the treatment of PF via inhibiting inflammation, oxidative stress, and endothelial mesenchymal transition, as well as affecting TGF-ß-mediated cell signaling, etc. Based on the current review, we have revealed the signaling mechanisms of PF pathogenesis and the potential opportunities offered by natural product-based medicine in treating PF.

Magnetoresistance Properties of Red Blood Cells in Plasma Combined with Several Magnetic Beads Passing Two Cu Electrodes.

In recent years, research has been intensively carried out on the applicability of magnetic beads (MBs) and magnetic nanoparticles coupled to biological objects such as red blood cells (RBCs). The magnetoresistance (MR) of a solution of RBCs and MBs (RBCs+MBs) was evaluated when MBs migrated in the presence or absence of an external magnetic field. The pattern of distribution of the MBs, which were homogeneously suspended in deionized distilled water, varied depending on the magnitude of the external magnetic field applied between the Cu electrodes connected to the two terminals. As the magnitude of the external magnetic field is increased or decreased, MBs are split on both sides and evenly mixed, respectively. The ratios (ΔMR/MR) versus an external magnetic field for the solutions of only MBs and a mixed RBCs+MBs were -33.4% and -27.4% at ±30 Oe and ±46 Oe of coercive fields, respectively. These results show that a solution of RBCs+MBs can act like a high-resolution biosensor that detects the oxygenation state of RBCs.

Gene Expression of Prox-1 and Hif-1a in Primo Vessels Inside Lymph Vessels of the Rabbit.

The gene expression of Prox-1 and Hif-1a for the isolated primo vessels (PVs) and composite lymphatic vessels (LVs) containing PVs (LVs + PVs) was investigated by RNA-sequencing (Seq) and quantitative polymerase chain reaction (qRT-PCR) analysis. RNA-Seq on the passed 10 samples on RNA-QC for two experimental groups with PVs and PVs + LVs proceeded to the library construction stage automatically and analyzed differentially expressed genes (DEGs). From the real-time qRT-PCR analysis data, we found the marker genes of Prox-1 and Hif-1a were enriched and decreased in an isolated PVs compared to LVs, respectively. Based on mRNA transcriptional data, Prox-1 and Hif-1a were increased and decreased in PVs compared to LVs + PVs under lipopolysaccharide (LPS) treatment and relieved by acupuncture electric stimulation (AES), respectively. This finding indicates that high and low levels of Prox-1 and Hif-1a may be involved in the function of PVs and that pathophysiological and physiological condition could progress into inflamed lymphatic endothelial cells expanding the PV within the LV.

Metabolomic analysis of organic acids, amino acids, and fatty acids in plasma of Hanwoo beef on a high-protein diet.

INTRODUCTION: Ketoacidosis of metabolic disease showed in beef cattle although body weight was increased by high-grain diets (HGDs). However, few studies have examined for health status related with metabolic disease of ketoacidosis following high-protein diet (HPD). OBJECTIVES: Metabolomic analysis was performed for the monitoring of health status associated with metabolic disease of ketoacidosis in the plasma of Hanwoo heifers following a HPD. METHODS: Hanwoo heifers of 24 months with 459 ± 42 kg weight were used under a 2 × 2 crossover design. The plasma was collected from control (n = 5) and HPD group (n = 5) on day 21 following diet adaptation for 20 days. Metabolic profiling analysis of organic acids (OAs), amino acids (AAs) and fatty acids (FAs) by gas chromatography-tandem mass spectrometry combined with star pattern analysis was performed in plasma. Levels of OAs, AAs and FAs were evaluated by Mann-Whitney test, PCA and PLS-DA. RESULTS: In HPD group, ketoacidosis as metabolic disease was monitored by elevated acetoacetic acid and 3-hydroxybutyric acid. In addition, the elevation of ketogenic AAs, reduction of medium chain FAs and OAs with energy metabolism in TCA cycle were monitored in HPD group. Star graphic pattern was characteristic and readily distinguished between control and HPD groups. In PLS-DA, two groups were separated with VIP score of top-ranked 10 FAs as important metabolites for discrimination. CONCLUSION: Elevation of ketone body including ketogenic AAs and reduced energy metabolism of FAs and OAs may useful for evaluation of health states associated with ketoacidosis from metabolic event by HPD in beef cattle.

Aestuariirhabdus litorea gen. nov., sp. nov., isolated from a sea tidal flat and proposal of Aestuariirhabdaceae fam. nov.

A Gram-negative, moderately halophilic and facultatively aerobic bacterium, designated strain GTF13T, was isolated from a sea tidal flat. Cells were curved rods and motile by a single polar flagellum showing catalase and oxidase activities. Growth was observed at 20-37 °C, pH 5.0-8.5 and 1.0-6.0 % (w/v) NaCl. Strain GTF13T contained C16:0, summed feature 3 (comprising C16 : 1 ω6c/C16 : 1 ω7c), summed feature 8 (comprising C18 : 1 ω6c/C18 : 1 ω7c) and C12 : 0 3-OH as major fatty acids and ubiquinone-9 and ubiquinone-8 as major quinones. Phosphatidylethanolamine and two unidentified phospholipids were detected as major polar lipids. The G+C content of the genomic DNA was 59.8 mol%. Strain GTF13T was most closely related to Simiduia agarivorans SA1T, Endozoicomonas montiporae CL-33T and Pseudomonas segetis FR1439T, belonging to different families or orders of the class Gammaproteobacteria, with less than 92.0 % 16S rRNA gene sequence similarities. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain GTF13T formed a phylogenetic lineage with the family Litoricolaceae, but the genome-based phylogenomic tree showed that strain GTF13T formed a distinct phylogenetic lineage within the order Oceanospirillales. The very low 16S rRNA gene sequence similarities and distinct phylogenetic relationships, together with distinct phenotypic and chemotaxonomic properties, served to differentiate strain GTF13T from phylogenetically closely related families. Here, strain GTF13T is proposed as a novel genus and species, for which the name Aestuariirhabdus litorea gen. nov., sp. nov. is proposed, within a new family Aestuariirhabdaceae fam. nov. of the order Oceanospirillales. The type strain is GTF13T (=KACC 19788T=JCM 32043T).

Geomonas soli gen. nov., sp. nov., a New Member of the Family Comamonadaceae, Isolated from Soil.

A Gram-stain negative, facultative aerobic bacterial strain, designated strain S-16T, was isolated from soil in South Korea. Colonies were white-milkish and cells were non-motile rods with oxidase- and catalase-positive activities. The growth of strain S-16T was observed at 20-40 °C (optimum, 25-30 °C) and pH 5.5-7.0 (optimum, pH 6.5). Ubiquinone-8 was identified as the sole respiratory quinone and C12:0, C16:0, C18:0, C15:1ω5c and summed feature 3 (comprising C16:1ω7c and/or C16:1ω6c) were identified as the major fatty acids (>5%). The major polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, an unidentified aminophospholipid, two unidentified phospholipids and an unidentified polar lipid. The G + C content of the genomic DNA calculated from the whole genome sequence was 66.8 mol%. Strain S-16T was most closely related to Piscinibacter aquaticus IMCC1728T, Rhizobacter gummiphilus NS21T and Rhizobacter dauci H6T with 16S rRNA gene sequence similarities of 97.93%, 97.93% and 97.44%, respectively. Phylogenetic analyses based on 16S rRNA gene and whole genome sequences suggested that strain S-16T could form a distinct phyletic lineage as a new genus within the family Comamonadaceae. Based on the phenotypic, chemotaxonomic and molecular features, strain S-16T represents the type strain of a novel species of a novel genus within the family Comamonadaceae, for which the name Geomonas soli gen. nov., sp. nov. is proposed. The type strain is S-16T (= KACC 19792T = JCM 32971T).

Analysis and Differential Expression of Primo Genes Using RNA-Seq and qRT-PCR Experiments.

Although the existence of the primo vasculature system has been shown in many species, including mice, rats, rabbits and humans, the biological role of this system, including expression of genes and proteins, has not yet been investigated. Especially the transcriptional action by mRNA, which is required for biological action, needs to be studied in primo vasculature biology. Differentially expressed genes in both isolated primo vessels and lymphatic vessels of rabbits were analyzed by RNA sequencing experiments. Primer efficiency and RNA purity of the primo vessels under lipopolysaccharides were confirmed prior to performing real-time qRT-PCR analysis following RNA extraction. We demonstrated that FLT4 was enriched in primo vessels and that several genes, including HSPH1 and EPHB2, were highly expressed in primo vessels compared with lymphatic vessels. Our data show that almost all genes, except HSPA4, were increased or sustained in isolated primo vessels compared with lymphatic vessels (FLT4 2.58 fold, HSPH1 1.83 fold, EPHB2 1.52 fold; whereas HSPA4 decreased 0.50 fold), suggesting primo vessels as a central regulator in diverse physiology. This implies that FLT4, HSPH1, and EPHB2 in high amounts may be involved in the functional activity of primo vessels. Our experimental data show that several genes are highly enriched in primo vessels in the lymphatic vessels of the rabbit.

Effect of γ-aminobutyric acid producing bacteria on in vitro rumen fermentation, growth performance, and meat quality of Hanwoo steers.

OBJECTIVE: The present study aimed to evaluate the effects of γ-aminobutyric acid (GABA)- producing bacteria (GPB) on in vitro rumen fermentation and on the growth performance and meat quality of Hanwoo steers. METHODS: The effects of GPB (Lactobacillus brevis YM 3-30)-produced and commercially available GABA were investigated using in vitro rumen fermentation. Using soybean meal as a substrate, either GPB-produced or commercially available GABA were added to the in vitro rumen fermentation bottles, as follows: control, no additive; T1, 2 g/L GPB; T2, 5 g/L GPB; T3, 2 g/L autoclaved GPB; T4, 5 g/L autoclaved GPB; T5, 2 g/L GABA; and T6, 5 g/L GABA. In addition, 27 Hanwoo steers (602.06±10.13 kg) were subjected to a 129-day feeding trial, during which they were fed daily with a commercially available total mixed ration that was supplemented with different amounts of GPB-produced GABA (control, no additive; T1, 2 g/L GPB; T2, 5 g/L GPB). The degree of marbling was assessed using the nine-point beef marbling standard while endotoxin was analyzed using a Chromo-Limulus amebocyte lysate test. RESULTS: In regard to in vitro rumen fermentation, the addition of GPB-produced GABA failed to significantly affect pH or total gas production but did increase the ammonia nitrogen (NH3-N) concentration (p<0.05) and reduce total biogenic amines (p<0.05). Animals fed the GPB-produced GABA diet exhibited significantly lower levels of blood endotoxins than control animals and yielded comparable average daily gain, feed conversion ratio, and beef marbling scores. CONCLUSION: The addition of GPB improved in vitro fermentation by reducing biogenic amine production and by increasing both antioxidant activity and NH3-N production. Moreover, it also reduced the blood endotoxin levels of Hanwoo steers.

Reduction of slaughter age of Hanwoo steers by early genotyping based on meat yield index.

OBJECTIVE: This study was conducted to determine early hereditary endowment to establish a short-term feeding program. METHODS: Hanwoo steers (n = 140) were equally distributed into four groups (35/group) based on genetic meat yield index (MYI) viz. the greatest, great, low, and the lowest at Jukam Hanwoo farm, Goheung. All animals were fed in group pens (5 animals/pen) with similar feed depending on the growth stage. Rice straw was provided ad libitum, whereas concentrate was fed at 5.71 kg during the growing period (6 to 13 mo) and 9.4 kg during the fattening period (13 to 28 mo). Body weight (BW) was measured at two-month intervals, whereas carcass weight was determined at slaughtering at about 31 months of age. The Affymetrix Bovine Axiom Array 640K single nucleotide polymorphism (SNP) chip was used to determine the meat quantity-related gene in the blood. RESULTS: After 6 months, the highest (p<0.05) BW was observed in the greatest MYI group (190.77 kg) and the lowest (p<0.05) in the lowest MYI group (173.51 kg). The great MYI group also showed significantly (p<0.05) higher BW than the lowest MYI group. After 16 and 24 months, the greatest MYI group had the highest BW gain (p<0.05) and were therefore slaughtered the earliest. Carcass weight was significantly (p<0.05) higher in the greatest and the great MYI groups followed by the low and the lowest MYI groups. Back-fat thickness in the greatest MYI group was highly correlated to carcass weight and marbling score. The SNP array analysis identified the carcass-weight related gene BTB-01280026 with an additive effect. The steers with the allele increasing carcass weight had heavier slaughter weight of about 12 kg. CONCLUSION: Genetic MYI is a potential tool for calf selection, which will reduce the slaughter age while simultaneously increasing carcass weight, back-fat thickness, and marbling score.

Recent insight and future techniques to enhance rumen fermentation in dairy goats.

Recent development of novel techniques in systems biology have been used to improve and manipulate the rumen microbial ecosystem and gain a deeper understanding of its physiological and microbiological interactions and relationships. This provided a deeper insight and understanding of the relationship and interactions between the rumen microbiome and the host animal. New high-throughput techniques have revealed that the dominance of Proteobacteria in the neonatal gut might be derived from the maternal placenta through fetal swallowing of amniotic fluid in utero, which gradually decreases in the reticulum, omasum, and abomasum with increasing age after birth. Multi "omics" technologies have also enhanced rumen fermentation and production efficiency of dairy goats using dietary interventions through greater knowledge of the links between nutrition, metabolism, and the rumen microbiome and their effect in the environment. For example, supplementation of dietary lipid, such as linseed, affects rumen fermentation by favoring the accumulation of α-linolenic acid biohydrogenation with a high correlation to the relative abundance of Fibrobacteriaceae. This provides greater resolution of the interlinkages among nutritional strategies, rumen microbes, and metabolism of the host animal that can set the foundation for new advancements in ruminant nutrition using multi 'omics' technologies.

Use of a Turn Coil and Channel Above a GMR-SV Device to Observe and Measure the Properties of Deoxidized Red Blood Cells Coupled to Magnetic Beads.

A giant magnetoresistance-spin valve (GMR-SV) having highly sensitive magnetic properties was prepared with a multilayer soft ferromagnetic/non-magnetic/pinned ferromagnetic/anti-ferromagnetic iridium manganese (IrMn) top layer (NiFe/CoFe/Cu/CoFe/IrMn) structure. Micron-sized magnetic beads (MBs) with a diameter of 1 µm immersed in a solution containing 50 mg of Co-Si-OH/ml were attached to red blood cells (RBCs) having the very low magnetization of hemoglobin (HEM) to detect a deoxidized RBC combined with MBs (RBC+MBs). The RBC+MB combinations can be captured on a single-turn µ-coil that maintains a magnetic field large enough to detect the MBs attached to the RBC. When the RBC+MBs passes through a PR µ-channel with a width of 10 µm, its stopping and starting are controlled by the electrical AC input signal applied to the single-turn µ-coil. The RBC+MBs combinations captured above the µ-device of the GMR-SV change the output signal and thus, can be used for detection. This observation implies that this device can be used as a biosensor to analyze the coupling force between the HEM and the MBs for RBCs with deformed membranes passing through a narrow capillary.

Effects of Rubus coreanus byproducts on intestinal microbiota and the immune modulation.

OBJECTIVE: Although the efficacy of Rubus coreanus (RC) byproducts as a feed additive has been recognized, its effects on intestinal microorganisms and the immune system are still unknown. METHODS: Six-week-old male rats were treated with 0.5% RC (T1), 1.0% RC (T2), and 1.5% RC (T3) for 4 weeks. RESULTS: We found that treatment with RC byproducts significantly increased the daily gain of body weight and feed intake. Treg-cell differentiation was enhanced in the mesenteric lymph nodes and spleen from the rats fed with RC byproducts. Illumina sequencing showed that bacteria in the phylum Firmicutes decreased and while those in the phylum Bacteroidetes increased in RC-treated groups. Particularly, the pathogenic microorganisms in the family Peptococcaceae decreased, and the non-pathogenic families Lachnospiraceae and S24-7 increased. Quantitative polymerase chain reaction analysis showed that the RC byproducts increased the lactic acid bacteria Bifidobacterium spp., Oscillospira spp., Leuconostoc citreum, and Weissella cibaria in a concentration-dependent manner. CONCLUSION: RC byproducts may be effective in immunomodulation by affecting intestinal microorganisms.

Effect of sodium stearoyl-2-lactylate supplementation on lactation performance, blood-biochemical profile, and economic efficacy of mid-lactation Holstein cows.

OBJECTIVE: This study was done to evaluate the effect of sodium stearoyl-2-lactylate (SSL) supplementation in a total mixed ration (TMR) on the lactation performance, blood parameters, and economic efficacy of mid-lactation Holstein cows. METHODS: Twenty-four cows (body weight 647±11.7 kg) were randomly divided into 4 treatment groups, with six cows per group. The dietary treatments were as follows: basal diet (CON); CON+17.5 g of top dressed SSL (treatment [TRT] 0.05); CON+35 g of SSL (TRT 0.1); and CON+70 g of SSL (TRT 0.2) per 35 kg TMR. RESULTS: The highest level of SSL supplementation (TRT 0.2) significantly improved milk yield during the second period compared to the TRT 0.05 group (5 to 8 wks; 33.28 vs 31.09 kg/d), during the third period compared to both the CON and TRT 0.05 groups (p<0.05) (9 to 13 wks; 32.59 vs 30.64 and 30.01 kg/d) and during the overall experimental period compared to both the CON and TRT 0.05 groups (p<0.05) (1 to 13 wks; 33.43 vs 32.06 and 31.40 kg/d), respectively. No negative effects on hematological or biochemical parameters were observed due to SSL supplementation. Considering both the milk fat and protein content, the total milk price was set at 1,073.60 (TRT 0.05), 1,085.60 (TRT 0.1), 1,086.10 (TRT 0.2), and 1,064.20 (CON) won/L, with consequent total milk profits of -1.7%, 5.4%, and 3.5% for the TRT 0.05, TRT 0.1, and TRT 0.2 diet, respectively, compared to those in the CON diet. CONCLUSION: The milk sales revenue related to SSL supplementation of the TRT 0.1 diet was increased by up to 5.4% compared to the milk sales revenue of the CON diet. Therefore, 0.1% SSL supplementation might be effective and profitable during the mid-lactation period of cows, without producing adverse effects.

Effect of optimal sodium stearoyl-2-lactylate supplementation on growth performance and blood and carcass characteristics in Hanwoo steers during the early fattening period.

OBJECTIVE: This study was conducted to evaluate the effect of different levels of total digestible nutrients (TDN) and sodium stearoyl-2-lactylate (SSL) supplementation on growth performance and blood and carcass characteristics in Hanwoo steers during the early fattening period. METHODS: Sixty Hanwoo steers (average body weight, 333±36.4 kg) were randomly allotted to 3 treatments, with twenty steers per treatment, and ten steers per pen with a size of 80 m2. Dietary treatments were as follows: CON, basal diet; treatment (TRT) 0.5, 0.5% down-spec of TDN with 0.1% SSL; TRT 1.0, 1.0% down-spec of TDN with 0.1% SSL. RESULTS: The results demonstrated that average daily gain and feed efficiency increased with TRT 0.5 (0.85 kg and 11.68) vs CON (0.82 kg and 11.27) or TRT 1.0 (0.78 kg and 10.74), indicating that 0.1% SSL supplementation in the feed of early fattening steers may result in a saving of 0.5% TDN. No significant differences were observed amongst all treatments (p> 0.05) for blood metabolite concentration and blood corpuscle values, which were all within the normally accepted range for healthy steers. CONCLUSION: Our study suggests that a TDN 0.5% down spec with 0.1% SSL supplemented feed may be effective and profitable for the early fattening period of Hanwoo steers without causing adverse effects.

Paenibacillus konkukensis sp. nov., isolated from animal feed.

A Gram-stain-positive, oxidase- and catalase-positive, aerobic, rod-shaped bacterium, designated strain SK-3146T, was isolated from animal feed. Phylogenetic analysis, based on 16S rRNA gene sequence comparisons, revealed that the strain formed a distinct lineage within the genus Paenibacillus that was closely related to Paenibacillusyunnanensis JCM 30953T (98.6 %), Paenibacillusvulneris CCUG 53270T (98.0 %) and Paenibacilluschinjuensis DSM 15045T (96.9 %). Cells were non-motile, endospore-forming and formed milky colonies on NA and R2A agar media. Growth of strain SK-3146T occurred at temperatures of 18-45 °C, at pH 6.0-9.5 and between 0.5-3.0 % NaCl (w/v). The major menaquinone was MK-7, with lesser amounts of MK-6 present. The cell wall peptidoglycan of strain SK-3146T contained meso-diaminopimelic acid. The major fatty acids were anteiso-C15 : 0 and iso-C16 : 0. The major polar lipids were diphosphatidylglycerol and phosphatidylethanolamine. The DNA G+C content was 53.8 mol% and the DNA-DNA hybridization relatedness values between strain SK-3146T and P.yunnanensis JCM 30953T and P.vulneris CCUG 53270T were 26.13±0.8 % and 38.7±0.6 %, respectively. The phenotypic, phylogenetic and chemotaxonomic results indicate that strain SK-3146T represents a novel species of the genus Paenibacillus, for which the name Paenibacillus konkukensis sp. nov. is proposed. The type strain is SK-3146T (=KACC 18876T=LMG 29568T).

Identification of the MUC2 Promoter as a Strong Promoter for Intestinal Gene Expression through Generation of Transgenic Quail Expressing GFP in Gut Epithelial Cells.

Identification of tissue- and stage-specific gene promoters is valuable for delineating the functional roles of specific genes in genetically engineered animals. Here, through the comparison of gene expression in different tissues by analysis of a microarray database, the intestinal specificity of mucin 2 (MUC2) expression was identified in mice and humans, and further confirmed in chickens by RT-PCR (reverse transcription-PCR) analysis. An analysis of cis-acting elements in avian MUC2 gene promoters revealed conservation of binding sites, within a 2.9 kb proximal promoter region, for transcription factors such as caudal type homeobox 2 (CDX2), GATA binding protein 4 (GATA4), hepatocyte nuclear factor 4 α (HNF4A), and transcription factor 4 (TCF4) that are important for maintaining intestinal homeostasis and functional integrity. By generating transgenic quail, we demonstrated that the 2.9 kb chicken MUC2 promoter could drive green fluorescent protein (GFP) reporter expression exclusively in the small intestine, large intestine, and ceca. Fluorescence image analysis further revealed GFP expression in intestine epithelial cells. The GFP expression was barely detectable in the embryonic intestine, but increased during post-hatch development. The spatiotemporal expression pattern of the reporter gene confirmed that the 2.9 kb MUC2 promoter could retain the regulatory element to drive expression of target genes in intestinal tissues after hatching. This new transgene expression system, using the MUC2 promoter, will provide a new method of overexpressing target genes to study gene function in the avian intestine.

Use of Lysozyme as a Feed Additive on In vitro Rumen Fermentation and Methane Emission.

This study was conducted to determine the effect of lysozyme addition on in vitro rumen fermentation and to identify the lysozyme inclusion rate for abating methane (CH4) production. An in vitro ruminal fermentation technique was done using a commercial concentrate to rice straw ratio of 8:2 as substrate. The following treatments were applied wherein lysozyme was added into 1 mg dry matter substrate at different levels of inclusion: Without lysozyme, 2,000, 4,000, and 8,000 U lysozyme. Results revealed that, lysozyme addition had a significant effect on pH after 24 h of incubation, with the highest pH (p<0.01) observed in 8,000 U lysozyme, followed by the 4,000 U, 2,000 U, and without lysozyme. The highest amounts of acetic acid, propionic acid (p<0.01) and total volatile fatty acid (TVFA) (p<0.05) were found in 8,000 U after 24 h of incubation. The CH4 concentration was the lowest in the 8,000 U and the highest in the without lysozyme addition after 24 h of incubation. There was no significant differences in general bacteria, methanogen, or protozoan DNA copy number. So far, addition of lysozyme increased the acetate, propionate, TVFA, and decreased CH4 concentration. These results suggest that lysozyme supplementation may improve in vitro rumen fermentation and reduce CH4 emission.

Characteristics of Wet Distillers Grains on In vitro Ruminal Fermentation and Its Effects on Performance and Carcass Characteristics of Finishing Hanwoo Steers.

Two experiments were conducted to determine the nutrient composition, in vitro ruminal ammonia concentrations and pH of wet distillers grains (WDG, produced from tapioca 70% and rice 30%) and to evaluate dietary effects of fermented total mixed ration (TMR) using WDG on the performance, blood metabolites and carcass characteristics of Hanwoo steers from mid fattening to slaughter. In Exp. I, average dry matter (DM), crude protein, ether extract, crude fiber, ash, neutral detergent fiber, acid detergent fiber, and nitrogen free extract of seven WDG samples from an ethanol plant with different sampling dates were 19.9%, 24.8%, 3.8%, 21.8%, 8.87%, 60.3%, 34.5%, and 40.7% (DM basis), respectively. For in vitro ammonia concentrations and pH, each sample was assigned to 7 incubation times (0, 4, 8, 12, 24, 48, and 72 h). Linear increase was observed between 12 and 48 h for ammonia concentrations, but final ammonia concentrations (72 h) were not significantly different among WDG samples and fermentation patterns of WDG samples showed similar tendency. In vitro pH varied among treatments from 0 to 24 h, but were not different statistically after 48 h. In Exp. II, 45 Hanwoo steers of 23 months (641±123 kg) from mid fattening period to slaughter (248 days) were randomly divided into three groups of 15 pens each (five repetitions/each treatment) and assigned to one of three dietary treatments; i) Control (TMR), ii) WDG 15 (TMR containing 15% of WDG, as fed basis) and iii) WDG 28 (TMR containing 28% of WDG, as fed basis). The body weight (BW), ADG, and feed conversion ratio (FCR) of control and WDG 15 and 28 during 248 days were 760.8, 740.1, and 765.5 kg, and 0.50, 0.50, and 0.52 kg/d, and 18.6, 17.6, and 17.1, respectively. The dry matter intake (DMI) (kg/d) of control (9.11) was higher (p<0.05) than WDG treatments (WDG 15%, 8.57; 28%, 8.70). Nevertheless, DMI did not affect BW, ADG, and FCR of Hanwoo finishing steers. Blood metabolites were in normal ranges and were not different among treatments except the albumin concentration. In carcass characteristics, WDG 15 (30%) showed higher frequency of A-carcass yield grade than WDG 28 (15%) and control (7%), and WDG 28 (61%) showed higher frequency of 1(++) and 1(+)-carcass quality grade than WDG 15 (40%) and control (60%). In conclusion, using WDG up to 28% in TMR did not show any negative effect on the performance and blood metabolites, and improved carcass quality of Hanwoo steers. Therefore, WDG can be a useful feed ingredient for Hanwoo steers in mid-fattening period to slaughter.