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Development of Non-toxic Recombinant Ricin Vaccine and Evaluation of Vaccine Efficacy (독성을 제거한 재조합 리신 백신 개발 및 효능평가)

  • Hyeongseok Yun;Hae-Eun Joe;Dong Hyun Song;Chi Ho Yu;Young-Jo Song;Gyeung Haeng, Hur
    • Journal of the Korea Institute of Military Science and Technology
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    • v.27 no.2
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    • pp.304-310
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    • 2024
  • Ricin is a highly toxic protein which is produced in the seeds of the castor oil plant. Ricin toxin A chain has ribosomal RNA N-glycosylase activity that irreversibly hydrolyses the N-glycosidic bond of the adenine residue at position 4324 within the 28S rRNA. In this study, we developed non-toxic recombinant ricin vaccine(R51) in E. coli expression system, and evaluated efficacy of the R51 according to adjuvants. When the R51 was administered using aluminum hydroxide as an adjuvant, the vaccine efficacy was higher than that of TLR agonists or aluminum phosphate. Because it is time-consuming to administer the vaccine three times at three-week intervals, we investigated the survival rate and antibody titer of mice according to the change of time interval of vaccination. Interestingly, there was no difference in survival rate and antibody titer when R51 was administered at 0, 1, and 3 weeks or 0, 2, and 4 weeks compared to when administered at 0, 3, and 6 weeks. Therefore, the developed R51 vaccine is promising to protect soldiers from Ricin attack.

Generation of ints14 Knockout Zebrafish using CRISPR/Cas9 for the Study of Development and Disease Mechanisms

  • Ji Hye Jung;Sanghoon Jeon;Heabin Kim;Seung-Hyun Jung
    • Development and Reproduction
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    • v.27 no.4
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    • pp.205-211
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    • 2023
  • INTS14/VWA9, a component of the integrator complex subunits, plays a pivotal role in regulating the fate of numerous nascent RNAs transcribed by RNA polymerase II, particularly in the biogenesis of small nuclear RNAs and enhancer RNAs. Despite its significance, a comprehensive mutation model for developmental research has been lacking. To address this gap, we aimed to investigate the expression patterns of INTS14 during zebrafish embryonic development. We generated ints14 mutant strains using the CRISPR/Cas9 system. We validated the gRNA activity by co-injecting Cas9 protein and a single guide RNA into fertilized zebrafish eggs, subsequently confirming the presence of a 6- or 9-bp deletion in the ints14 gene. In addition, we examined the two mutant alleles through PCR analysis, T7E1 assay, TA-cloning, and sequencing. For the first time, we used the CRISPR/Cas9 system to create a model in which some sequences of the ints14 gene were removed. This breakthrough opens new avenues for in-depth exploration of the role of ints14 in animal diseases. The mutant strains generated in this study can provide a valuable resource for further investigations into the specific consequences of ints14 gene deletion during zebrafish development. This research establishes a foundation for future studies exploring the molecular mechanisms underlying the functions of ints14, its interactions with other genes or proteins, and its broader implications for biological processes.

Effect of phytase supplementation on performance, fecal excretion, and compost characteristics in broilers fed diets deficient in phosphorus and calcium

  • Chun Ik Lim;Hyo Jun Choo;Jae Hong Park
    • Journal of Animal Science and Technology
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    • v.66 no.1
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    • pp.93-102
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    • 2024
  • This study investigated the effect of dietary supplementation with phytase on growth performance, fecal excretion, and compost nutrition on broilers fed available phosphorus (avP)- and calcium (Ca)-deficient diets. A total of 750 one-day-old broiler chicks were randomly divided into five dietary groups having ten replications in a floor house. Diets of the groups were formulated with positive control (PC), negative control (NC; low avP and Ca), and NC supplemented with phytase levels; 500 (NC500), 1,000 (NC1000), and 1,500 FTU/kg (NC1500). A three-phase feeding program was used in the trial. Average daily gain (ADG) and average daily feed intake (ADFI) in the groups fed diets supplemented with phytase were significantly (p < 0.05) higher than those fed NC and the increase was equivalent to those fed PC. Serum levels of Ca and phosphorus (P) were higher (p < 0.05) in broilers fed NC1000 and NC1500 than in those fed NC. Interleukin (IL) level was the lowest in the group fed NC. Plasma myo-inositol (INS) concentrations in the NC1500 group were higher (p < 0.05) than PC, NC, and NC500 groups. Crude protein (CP) excretion was notably (p < 0.05) lower in the NC1500 group than in PC and NC groups. A lower (p < 0.05) concentration of P2O5 was observed in compost from the group fed NC1500 than the groups fed PC and NC. Accordingly, we suggest that phytase supplementation in lower avP and Ca levels of broiler diet can improve their productive performance and reduce environmental pollution.

Effects of supplemental bacteriophage on the gut microbiota and nutrient digestibility of ileal-cannulated pigs

  • Hyunwoong Jo;Geongoo Han;Eun Bae Kim;Changsu Kong;Beob Gyun Kim
    • Journal of Animal Science and Technology
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    • v.66 no.2
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    • pp.340-352
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    • 2024
  • This study measured the potential changes of the microbiota in the gastrointestinal tract and energy and nutrient digestibility by supplemental bacteriophages in pigs. Twelve castrated male pigs (initial mean body weight = 29.5 ± 2.3 kg) were surgically cannulated using T-cannula. The animals were housed individually in pens equipped with a feeder and a nipple waterer. The pigs were allotted to 1 of 3 experimental diets in a quadruplicated 3 × 2 Latin square design with 3 experimental diets, 2 periods, and 12 pigs resulting in 8 replicates per diet. The 3 diets were a control mainly based on corn and soybean meal with no antibiotics or bacteriophages, a diet containing 0.1% antibiotics, and a diet containing 0.2% bacteriophages. On day 5 of the experimental period, feces were collected and on days 6 and 7, ileal digesta were collected. Genomic DNA for bacteria were extracted from the ileal digesta and feces and the V4 region of the 16S rRNA gene was amplified. The ileal and fecal digestibility of energy, dry matter, organic matter, crude protein, and fiber was unaffected by dietary antibiotics or bacteriophages. At the phylum level, the supplemental antibiotic or bacteriophage tended to result in a higher proportion of Firmicutes (p = 0.059) and a lower proportion of Bacteroidetes (p = 0.099) in the ileal digesta samples compared with the control group with no difference between the antibiotic and bacteriophage groups. At the genus level, the supplemental antibiotic or bacteriophage tended to result in a higher proportion of Lactobacillus (p = 0.062) and a lower proportion of Bacteroides (p = 0.074) and Streptococcus (p = 0.088) in the ileal digesta compared with the control group with no difference between the antibiotic and bacteriophage groups. In the feces, supplemental antibiotics or bacteriophages reduced the proportion of Bifidobacterium compared with the control group (p = 0.029) with no difference between the antibiotic and bacteriophage groups. Overall, supplemental antibiotics and bacteriophages showed positive effect on the microbiota of in the ileal digesta without largely affecting energy or nutrient digestibility, with no differences between the antibiotic and bacteriophage groups in growing pigs.

Comparison study between single enzyme and multienzyme complex in distiller's dred grains with soluble supplemented diet in broiler chicken

  • Min-Jin Kwak;Dong-Jin Ha;Min Young Park;Ju Young Eor;Kwang-Youn Whang;Younghoon Kim
    • Journal of Animal Science and Technology
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    • v.66 no.2
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    • pp.398-411
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    • 2024
  • Upregulation of the nutritional value of feed is the major target of various studies in the livestock industry, and dietary enzyme supplementation could aid in digesting the nondegrading nutrients of grains in feed ingredients. Dried distillers' grains with solubles (DDGS) is a byproduct of the fermentation process in the beverage industry and can be used as a large supply source of fiber in feed. Therefore, we conducted an experiment with male broiler chickens to investigate the effect of various types of enzymes on DDGS and compare the efficacy of single enzyme and multienzyme complexes on growth performance and gut environments in broiler chickens. We used 420 1-day-old broiler chickens (Ross 308), and they were allotted into 4 dietary treatments with seven replications (CON, corn-soybean meal [SBM] diet; NC, DDGS supplemented diet; SE, 0.05 % of mannanase supplemented DDGS-based diet; MC, 0.10% of multienzyme complex (mannanase and xylanase, glucanase) supplemented DDGS-based diet. The dietary exogenous enzyme in the DDGS-supplemented diet could improve growth performance as much as the growth of the control group, and digestibility of dry matter, crude protein, and gross energy were significantly increased by enzyme addition in groups of chicks fed DDGS-supplementation diet. Moreover, the populations of pathogenic bacteria, coliforms, and Bacteroidetes were significantly decreased by enzyme supplementation, which might lead to improved gut mucus-secreting cells and inflammatory cytokines in the jejunum. Collectively, dietary single enzyme and multienzyme complexes could improve gut environments, including intestinal immune responses and gut microbial population, and lead to improvement of growth performance in broiler chickens.

Lysophosphatidic acid improves development of porcine somatic cell nuclear transfer embryos

  • Ling Sun;Tao Lin;Jae Eun Lee;So Yeon Kim;Ying Bai;Dong Il Jin
    • Journal of Animal Science and Technology
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    • v.66 no.4
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    • pp.726-739
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    • 2024
  • This study was conducted to investigate whether lysophosphatidic acid (LPA) could improve the development of porcine somatic cell nuclear transfer (SCNT) embryos. Porcine SCNT-derived embryos were cultured in chemically defined polyvinyl alcohol (PVA)-based porcine zygote medium (PZM)-4 without or with LPA, and the development, cell proliferation potential, apoptosis, and expression levels of pluripotent markers were evaluated. LPA significantly increased the rates of cleavage and blastocyst formation compared to those seen in the LPA un-treatment (control) group. The expression levels of embryonic development-related genes (IGF2R, PCNA and CDH1) were higher (p < 0.05) in the LPA treatment group than in the control group. LPA significantly increased the numbers of total, inner cell mass and EdU (5-ethynyl-2'-deoxyuridine)-positive cells in porcine SCNT blastocysts compared to those seen in the control group. TUNEL assay showed that LPA significantly reduced the apoptosis rate in porcine SCNT-derived embryos; this was confirmed by decreases (p < 0.05) in the expression levels of pro-apoptotic genes, BAX and CASP3, and an increase (p < 0.05) in the expression level of the anti-apoptotic gene, BCL2L1. In addition, LPA significantly increased Oct4 expression at the gene and protein levels. Together, our data suggest that LPA improves the quality and development of porcine SCNT-derived embryos by reducing apoptosis and enhancing cell proliferation and pluripotency.

Complete genome sequence of candidate probiotic Limosilactobacillus fermentum KUFM407

  • Bogun Kim;Ji yu Heo;Xiaoyue Xu;Hyunju Lee;Duleepa Pathiraja;Jae-Young Kim;Yi Hyun Choi;In-Geol Choi;Sae Hun Kim
    • Journal of Animal Science and Technology
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    • v.66 no.4
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    • pp.859-862
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    • 2024
  • It has been reported that the administration of Limosilactobacillus fermentum alleviates diseases such as osteoporosis and colitis. In this study, we report the complete genome sequence of Limosilactobacillus fermentum KUFM407, a probiotic strain of LAB isolated from Korean traditional fermented food, Kimchi. Whole genome sequencing of L. fermentum KUFM407 was performed on the Illumina MiSeq and Oxford Nanopore MinION platform. The genome consisted of one circular chromosome (2,077,616 base pair [bp]) with a guanine cytosine (GC) content of 51.5% and one circular plasmid sequence (13,931 bp). Genome annotation identified 1,932 protein-coding genes, 15 rRNAs, and 58 tRNAs in the assembly. The function annotation of the predicted proteins revealed genes involved in the biosynthesis of bacteriocin and fatty acids. The complete genome of L. fermentum KUFM407 could provide valuable information for the development of new probiotic food and health supplements.

Potential use of dried persimmon (Diospyros kaki) byproducts as feed sources for ruminants

  • Sang Moo Lee;Tabita Dameria Marbun;Eun Joong Kim
    • Journal of Animal Science and Technology
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    • v.66 no.4
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    • pp.749-762
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    • 2024
  • The aim of this study was to evaluate the chemical composition, in vitro digestibility, and palatability of dried persimmon byproducts (persimmon peel [PP] and damaged whole persimmons [WP]) ensiled with rice straw in different mixing ratios. PP and WP were ensiled with rice straw at ratios of 3:7 (PP3R7, WP3R7), 5:5 (PP5R5, WP5R5), 7:3 (PP7R3, WP7R3), and 8:2 (PP8R2, WP8R2) for 70 d. WP3R7 had the highest (p < 0.05) crude protein and lactate contents compared to the other combinations. On the other hand, PP3R7 and PP8R2 had lower concentrations of neutral and acid-detergent fibers (p < 0.05) and produced lower amounts of ammonia-N (p < 0.05). The silages were compared to rice straw silage (RS), maize silage (MS), whole-crop rye silage (WCRS), and sorghum-sudangrass silage (SSGS) during an in vitro study. The results showed that PP8R2 and WP7R3 had higher (p < 0.05) dry matter digestibility values than RS, MS, WCRS, and SSGS in a 6 h incubation period. In addition, a palatability test of the silages was conducted on Hanwoo cattle, goats, and deer, using the cafeteria method. The palatability index rate of PP7R3 was the highest (p < 0.05) for the goats and the Hanwoo cattle, whereas PP8R2 had the highest (p < 0.05) rate for the deer and the Hanwoo cattle. In conclusion, dried persimmon byproducts in the form of PP and WPs can be used as ruminant feed when ensiled with RS at ratios of 7:3 and 8:2.

Rapamycin treatment during prolonged in vitro maturation enhances the developmental competence of immature porcine oocytes

  • Seung-Eun Lee;Han-Bi Lee;Jae-Wook Yoon;Hyo-Jin Park;So-Hee Kim;Dong-Hun Han;Eun-Seo Lim;Eun-Young Kim;Se-Pill Park
    • Journal of Animal Science and Technology
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    • v.66 no.5
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    • pp.905-919
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    • 2024
  • Porcine oocytes undergo in vitro maturation (IVM) for 42-44 h. During this period, most oocytes proceed to metaphase and then to pro-metaphase if the nucleus has sufficiently matured. Forty-four hours is sufficient for oocyte nuclear maturation but not for full maturation of the oocyte cytoplasm. This study investigated the influences of extension of the IVM duration with rapamycin treatment on molecular maturation factors. The phospho-p44/42 mitogen-activated protein kinase (MAPK) level was enhanced in comparison with the total p44/42 MAPK level after 52 h of IVM. Oocytes were treated with and without 10 µM rapamycin (10 R and 0 R, respectively) and examined after 52 h of IVM, whereas control oocytes were examined after 44 h of IVM. Phospho-p44/42 MAPK activity was upregulated the 10 R and 0 R oocytes than in control oocytes. The expression levels of maternal genes were highest in 10 R oocytes and were higher in 0 R oocytes than in control oocytes. Reactive oxygen species (ROS) activity was dramatically increased in 0 R oocytes but was similar in 10 R and control oocytes. The 10 R group exhibited an increased embryo development rate, a higher total cell number per blastocyst, and decreased DNA fragmentation. The mRNA level of development-related (POU5F1 and NANOG) mRNA, oocyte-apoptotic (BCL2L1) genes were highest in 10 R blastocysts. These results suggest that prolonged IVM duration with rapamycin treatment represses ROS production and increases expression of molecular maturation factors. Therefore, this is a good strategy to enhance the developmental capacity in porcine oocytes.

Development of semi-dried goat meat jerky using tenderizers considering the preferences of the elderly

  • Shine Htet Aung;Md. Altaf Hossain;Ji-Young Park;Young-Sun Choi;Ki-Chang Nam
    • Journal of Animal Science and Technology
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    • v.66 no.4
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    • pp.807-833
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    • 2024
  • Elderly people avoid eating red meat and dried meat product due to its texture and stiffness; they deprive them of rich sources of nutrients. In addition, food-related diseases are exponentially increasing due to using synthetic additives in food products. Therefore, this research aimed to develop semi-dried goat meat jerky considering geriatric preferences by using natural tenderizers and nitrate. Four treatments were formulated negative control (NC [synthetic nitrite without tenderizers]), positive control (PC [Swiss chard without tenderizers]), T1 (Swiss chard with pineapple powder), and T2 (Swiss chard with pineapple and tomato powder). T1 and T2 had higher processing yield, and rehydration capacity compared with NC and PC. The fat content of T1 and T2 was lower than the control groups. Moisture was significantly higher in T1, NC, and T2 than in PC (p < 0.05). T2 showed the lowest water activity (0.87), lowest shear force (4.82 kgf), and lowest total plate count (TPC). The lowest pH and thiobarbituric acid reactive substances (TBARS) were observed in T1 and T2. T1 showed the lowest lightness and the maximum redness (p < 0.05) while PC showed the lowest yellowness. During the storage period, moisture and pH decreased, and TPC and TBARS significantly increased whereas water activity is stable regardless of the treatment. The results of the myofibrillar fragmentation index (MFI) and sodium dodecyl sulfate-polyacrylamide gel revealed that T1 and T2 more effectively converted protein to polypeptides. In addition, tenderizers positively affected thrombogenicity, atherogenicity, and hypocholesterolemic/hypercholesterolemic indices. T2 observed the highest overall sensory acceptance by reducing goaty flavor. Overall, jerky treated with tenderizers is easily chewable and digestible for the elderly due to its tenderness and essential fatty acids that would be senior-friendly food.