• Asian-Australasian Journal of Animal Sciences
• /
• v.30 no.10
• /
• pp.1478-1485
• /
• 2017

Objective: The effects of vaccinating 18-day-old chicken embryos with the combination of recombinant Eimeria profilin plus Clostridium perfringens (C. perfringens) NetB proteins mixed in the Montanide IMS adjuvant on the chicken immune response to necrotic enteritis (NE) were investigated using an Eimeria maxima (E. maxima)/C. perfringens co-infection NE disease model that we previously developed. Methods: Eighteen-day-old broiler embryos were injected with $100{\mu}L$ of phosphate-buffered saline, profilin, profilin plus necrotic enteritis B-like (NetB), profilin plus NetB/Montanide adjuvant (IMS 106), and profilin plus Net-B/Montanide adjuvant (IMS 101). After post-hatch birds were challenged with our NE experimental disease model, body weights, intestinal lesions, serum antibody levels to NetB, and proinflammatory cytokine and chemokine mRNA levels in intestinal intraepithelial lymphocytes were measured. Results: Chickens in ovo vaccinated with recombinant profilin plus NetB proteins/IMS106 and recombinant profilin plus NetB proteins/IMS101 showed significantly increased body weight gains and reduced gut damages compared with the profilin-only group, respectively. Greater antibody response to NetB toxin were observed in the profilin plus NetB/IMS 106, and profilin plus NetB/IMS 101 groups compared with the other three vaccine/adjuvant groups. Finally, diminished levels of transcripts encoding for proinflammatory cytokines such as lipopolysaccharide-induced tumor necrosis $factor-{\alpha}$ factor, tumor necrosis factor superfamily 15, and interleukin-8 were observed in the intestinal lymphocytes of chickens in ovo injected with profilin plus NetB toxin in combination with IMS 106, and profilin plus NetB toxin in combination with IMS 101 compared with profilin protein alone bird. Conclusion: These results suggest that the Montanide IMS adjuvants potentiate host immunity to experimentally-induced avian NE when administered in ovo in conjunction with the profilin and NetB proteins, and may reduce disease pathology by attenuating the expression of proinflammatory cytokines and chemokines implicated in disease pathogenesis.

• Korean Journal of Poultry Science
• /
• v.16 no.1
• /
• pp.1-8
• /
• 1989

This experiment was conducted to improve the performance of chickens by the precise separation and analysis of chromosomes which are integrated genetic materials, and by the use of gene manipulation techniques. Following are the main results obtained. 1. When the chromosomes were separated through the leucocyte culture and analyzed by Giemsa banding techniques (especially by the method in which 20 layers of banding patterns could be found in chromosome ＃1), the normal Patterns of chromosomes ＃l-9 and sex chromosomes, and the location of constitutive heterochromatin without any gene activities in all chromosomes were discovered. 2. To utilize the primodial germ cells (PGC) as the genetic vector which is one of the most important gene manipulation techniques, PGC's from triploid were transplanted to normal host embryos. Since the donor PGC's(3n) were found in the gonads of growing host embryos gene manipulation in poultry using PGC's, seemed to be possible.

• The Korean Journal of Mycology
• /
• v.14 no.1
• /
• pp.61-70
• /
• 1986

Sterigmatocystin bears a close structural relationship to aflatoxin $B_1$ and is a carcinogenic compound that has been shown to affect various species of experimental animals. Reaction and toxicity of sterigmatocystin in the artificial gastric juice were investigated. Sterigmatocystin was degraded in artificial gastric juice and extracted by the method of A.O.A.C. After cleaned up by silica gel column chromatography, this substance was detected and characterized by thin layer chromatography, UV, IR and mass spectra. It showed $R{\mathcal{f}}$ 0.4 and brick-red color by TLC. Especially, in the mass spectrum of it, fragment peak at m/e 327 was due to the loss of the $-CH_3$ and $-H_2O$, fragment peak at m/e 341 was due to the loss of the $H_2O$ and $-H^+$, and fragment peak at m/e 239 was due to the loss of the 2-chloro-tetrahydrofuran and methyl group from the parent molecule. Therefore, a degraded substance of sterigmatocystin reacted in artificial gastric juice (Sub. K) was estimated with additional formation of hydrochloric acid. In four-day-old chicken embryos, the mean lethal dose $(LD_{50})$ was $140\;{\mu}g/egg$, and 90 to 100% of the embryos were killed with 1 mg/egg. This $LD_{50}$ $140\;{\mu}g/egg$ compared with an $LD_{50}$ $14.69\;{\mu}g/egg$ for sterigmatocystin (acute toxicity) showed the substance to be much less toxic than sterigmatocystin.

• Korean Journal of Poultry Science
• /
• v.23 no.1
• /
• pp.9-17
• /
• 1996

This study was conducted to compare the survival rate of chick embryos among different eggshell window positions and to search for the most appropriate injection position. The eggshells were punctured at blunt-end, sharp-end and side-up with a sterilized fine forceps, respectively. The survival rate of sharp-end window was higher than the other window positions. Injection of Dulbecco’s modified eagle’s medium (DMEM) through blunt-end window (BE1) was impossible because inner cell membrane was obscure. The 2 ${\mu}$L DMEM was injected into 2.5 d-old embryo blood vessel through sharp end window. To prevent hemorrhages at the point of injection, the air bubbles were injected into the embryo blood vessel. The survival rate of chicks embryo in sharp end window was about 17.0％. Therefore, this sharp-end window system will be helpful for the production of germline chimera or transgenic chicken using primordial germ cells ( PGCs ).

• Analytical Science and Technology
• /
• v.26 no.6
• /
• pp.401-406
• /
• 2013

Oxygen($O_2$) consumption and carbon dioxide($CO_2$) excretion of a fertile chicken egg during incubation were measured by a gas mass spectrometer. A closed sample chamber was developed to collect gas samples during the 20 days of artificial incubation of both a fertile and an infertile egg. After leaving an egg in the sample chamber for an hour, using a gas-tight syringe, samples of 2 mL of gas were collected from the closed sample chamber and analyzed using a gas mass spectrometer in 2~4 day intervals. The $O_2$ consumption and $CO_2$ excretion of chicken embryos increased rapidly after 10 days from the starting point of incubation. After 20 days, 23 mL of $O_2$ was consumed and 16 mL of $CO_2$ was excreted per hour. Throughout the whole period of incubation, concentration of $O_2$ decreased 4.3 mol% and $CO_2$ increased only 3.1 mole%, i.e., the mole of consumed $O_2$ and the mole of excreted $CO_2$ were not the same. On the other hand, during the same period, concentration of $N_2$ increased about 1.3 mol% and the increased mole fraction of $N_2$ was comparable with the difference (1.2 mol%) between the mole fraction of consumed $O_2$ and excreted $CO_2$. Therefore, we can attribute the increase of $N_2$ mole% to the difference of mole fraction between consumed $O_2$ and excreted $CO_2$. In this study, through the analysis of gas, we could explain the respiration of a fertile chicken egg during incubation.

• Proceedings of the Korea Society of Poultry Science Conference
• /
• 2005.11a
• /
• pp.66-67
• /
• 2005

The domestic chicken (Gallus gallus) is an important model for research in developmental biology because its embryonic development occurs in ovo. To examine the mechanism of embryonic germ cell development, we constructed proteome map of gonadal primordial germ cells (gPGC) from chicken embryonic gonads. Embryonic gonads were collected from 500 embryos at 6 day of incubation, and the gPGC were cultured in vitro until colony formed. After 7-10 days in cultured gPGC colonies were separated from gonadal stroma cells (GSCs). Soluble extracts of cultured gPGCs were then fractionated by two-dimensional gel electrophoresis (pH 4-7). A number of protein spots, including those that displayed significant expression levels, were then identified by use of matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry and LC-MS/MS. Of the 89 gPGC spots examined, 50 yielded mass spectra that matched avian proteins found in on-line databases. Proteome map of thistype will serve as an important reference for germ cell biology and transgenic research.

• Korean Journal of Poultry Science
• /
• v.38 no.2
• /
• pp.89-96
• /
• 2011

The outbreak of Newcastle disease occurred for the first time at a commercial chicken farm near Ulaanbaatar, Mongolia in August 2010. Newcastle disease virus (NDV) obtained from infected chickens in Mongolia was characterized by biological and molecular biological approches. Mongolian NDV isolate killed all of chicken embryos within 60 h in the mean death time assay, indicating virulent for chicken. A genomic region of 695 nts between nts 1055 of the M gene and 508 of the F gene was amplified by RT-PCR and sequenced. The deduced amino acid sequence of the F protein cleavage site was $^{112}RRQKRF^{117}$, which is a typical sequence of velogenic strains of NDV and is agreement with the result of the MDT assay. The sequence of the partial F gene (nts 47 to 435) was used for genotyping by phylogenetic analysis. The phylogenetic analysis showed that the Mongolian isolate was of genotype VII within class II of NDV. Further phylogenetic analysis on the genotype VII strains revealed that the isolates placed in a genetic sublineage of VIId and most closely related with velogenic strains of NDV circulating in Far-east Asian region especially China, suggesting the introduction of velogenic NDV into Mongolia from neighboring countries.

• Asian-Australasian Journal of Animal Sciences
• /
• v.25 no.9
• /
• pp.1229-1236
• /
• 2012

Our previous studies showed that kisspeptin-10 (Kp-10) injected in vivo can markedly increase lipid anabolism in liver of quails. In order to investigate the direct effect of Kp-10 on lipid metabolism of hepatocytes in birds, cells were separated from embryos livers and cultured in vitro with 0, 100 and 1,000 nM Kp-10, respectively. The results showed that after 24 h treatment, cells viability was not affected by 100 nM Kp-10, but showed a mild decrease with 1,000 nM Kp-10 compared to the control cells. Based on the results of the cell viability, 100 nM dosage of Kp-10 was selected for the further study and analysis. Compared with control cells, total cholesterol (Tch) contents in 100 nM treated cells were increased by 51.23%, but did not reach statistical significance, while the level of triglyceride (TG), high density of lipoprotein-cholesterol (HDL-C) and low density of lipoprotein-cholesterol (LDL-C) were significantly increased. Real-time PCR results showed that ApoVLDL-II mRNA expression had a tendency to increase, genes including sterol regulatory element-binding protein-1 (SREBP-1), acetyl coenzyme A carboxylase ${\alpha}$ ($ACC{\alpha}$), carnitine palmitoyltransferase 1 (CPT1), 3-hydroxyl-3-methylglutaryl-coenzyme A reductases (HMGCR) and stearyl coenzyme A dehydrogenase-1 (SCD1) mRNA in hepatocytes were significantly down-regulated by 100 nM Kp-10. However, contrary to its gene expression, SREBP-1 protein expression was significantly up-regulated by 100 nM Kp-10. Some of the significant correlations in mRNA expression were found between genes encoding hepatic factors or enzymes involved in lipid metabolism in liver of birds. These results indicate that Kp-10 stimulates lipid synthesis directly in primary cultured hepatocytes of chickens.

• Korean Journal of Veterinary Research
• /
• v.8 no.2
• /
• pp.98-109
• /
• 1968

The small type piroplasma which is known to be prevailing in Korean cattle was injected to un-susceptible hosts such as mice, guinea pigs, rats, rabbits, fowls and developing chicken eggs by i.v., i.p, and intra C.A.M. routes, and its morphological development in each animal species was observed. 1. The small type piroplasma of cattle did not produce any pathogenicity in the experimental animals such as mice, guinea pigs, rats, rabbits, fowls and developing chicken embryos. However, only a partial sign for developing cycle of the organism was observed in the peritoneal fluid. The schizogony of protozoa was not detected in any organs of the inoculated animals. 2. In the animals inoculated, the parasites of I-III types, relative to the process of time in the host, have changed their shapes into IV-VI types and then VI-X types however, the type X was not restored to the type I. In guinea pigs, more grape type was detected then X type in number, and in process of time, the shape was changed from round to elliptic, and the pointed end of the organism appeared in flesh color. 3. None of the organisms were found from the blood stream of inoculated animals excepts only a few piroplasma as were found in the experimental mice shortly after inoculation or i.v. route, No protozoa was also found in the organs of the animals autopsied. 4. The form change of protozoa was more clear in these animals than that in susceptible host. The most piroplasmas have changed their forms into VI-X types 30 hours after inoculation and were dispersed from R.B.C. Tracing of piroplasrna beyond this stage was not attempted. 5. The small type piroplasma comprises in I. X types and it is further believed that VII-X types which were freed from one cell proceeded to invade others. The from of development resembles to the small type of piroplasma which is prevailing in Japan.

• Korean Journal of Poultry Science
• /
• v.22 no.1
• /
• pp.31-42
• /
• 1995

This study was to investigate the effects of dietary linoleic acid(18:2\omega6, LA) and aipha-linolenic acid(18:3\omega3. \alpha-LNA) levels on brain development and fatty acid compositions of various lipid classes in the chicken embryo brain tissues. Thirty two ISA Brown layers, 52 weeks-old, were divided into four groups. Birds of each group were given corn-soybean meal based diets added with 1) safflower oil 8%, 2) safflower oil 6% + perilla oil 2%, 3) safflower oil 2% + perilla oil 6%, or 4) perilla oil 8%. Mter 15 days fed the diets. the layers were artificially inseminated to obtain fertile eggs. During the incubation. embryonic brains were sampled at 15th and 21st days. Fatty acid contents were quantitated by using heptadecanoic acid (17:0) as an internal standard. No significant differences in brain weight and in contents of various lipids such as phospholipid. triglyceride, cholesterol. cholesterol ester and free fatty acid in the tissues were found among the dietary groups (P<0.05). The ratios of AA/LA in the brain lipid classes were lowered as the dietary levels of perilla oil were increased. Higher LA was found in phosphatidylcholine(PC) than arachidonic acid (20:4\omega6. AA), meanwhile the level of LA was less than AA in phosphatidylethanolamine(PE). Docosahexaenoic acid(22:6\omega3, DHA) was the* major fatty acid in the tissue and its content in PE was 2.5~3 times higher than in PC. DHA level in the phospholipid reached at a peak (1.7~1.8 mg/brain) in dietary groups added with 6% or 8% perilla oil. suggesting that no more increase in that fatty acid level in the brain tissue could be obtained by consuming more \alpha-LNA, the major precursor of DHA.