열 안정성을 가지고 있는 한국산 꿩에서 분리된 New-castle disease virus CBP-1주는 9일령 SPF 계태아에 접종되어 $37^{\circ}C$에서 배양하는 방법으로 173번(parent주) 누대 배양되었다. $37^{\circ}C$에서 173번 누대 배양된 NDV CBP-1 주를 10 일령 계태아에 접종한 후 저온에서 ($29^{\circ}C$) 15번 (CA-15) 30번(CA-30) 누대 배양하였다. 저온순화 주인 CA-15주와 CA-30주의 이화학적 성상검사 (열 안정성 실험, 지질 용매에 대한 감수성 실험, 산성 용매에 대한 감수성 실험)와 병원성 실험(MDT, ICPI, IVPI), 온도 감수성 실험, 안전성 실험, 부스터 효과 실험, 방어효과 등을 실험하였고, $37^{\circ}C$에서 173번 누대 배양된 parent주와 비교하였다. $29^{\circ}C$에 적응된 CA-30주는 $37^{\circ}C$와 $41^{\circ}C$에서 세포 감염력이 parent주와 비교할 때 감소하였다. CA-15주와 CA-30주를 $56^{\circ}C$에서 30분, 60분, 120분 동안 처리하였을 때 이들 저온 순화주들은 혈구응집능과 세포감염력을 상실하였다. Parent주와 CA-15, CA-30주는 ethyl ether를 10분간 처리했을 때 혈구응집능과 세포 감염력을 모두 상실하였다. 그러나 parent주와 CA-15, CA-30주는 pH 3.0-glycine HCl 완충액에 60분간 처리하였을 때 혈구응집능을 가지고 있었다. Parent주의 대뇌병원성 지수와 정맥내 병원성 지수는 각각 1.12, 1.45 이었다. 그러나 CA-30주의 대뇌 병원성 지수와 정맥내 병원성 지수는 각각 0.75, 0.00으로 감소하였다. CA-30주의 안전성은 1 일령 병아리에서 실시하였고 parent주와 B-1주와 비교였다. 이들의 안전성은 치사율을 가지고 평가하였다. parent주와 CA-30주와 B-1주의 치사율은 각각 17.5, 12.0. 0.0%이었다. CA-30주가 parent주에 비해 보다 높은 안전성을 보여 주었으나 B-1주에 비해서 아직 높은 치사율을 보여 주고 있었다.

소나무 수피로부터 alkaline sulfite-anthraquinone cooking을 통하여 생산된 Pine Bark Spent Liquor(PBSL)의 펠렛 내구성(ellet durability index(PDI)과 사료결착재로서 가능성을 구명하기 위하여 육계와 산란계 사료에 첨가·급여하였다. 실험1에서는 실용계 사료인 대조구, 소맥 10%와 (W10), 20% 첨가구(W20), PBSL 0.25, 0.50, 1.00, 2.00, 3.00% 첨가구, 두 종류 시판용 사료결착제 0.1, 0.2%(CPB A와 CPB B), W10+0.1% CPB A or B로 14개 처리구를 4반복으로 하여 펠렛 내구성 조사를 시행하였다. PDI는 PDI tester(Oriental motors, Japan)로 측정하였다. 실험2에서는 1일령 육계수컷 200수를 무첨가구인 대조구외에 실험 1에서 최적으로 판단되는 처리구인 W10, PBSL 0.5%, 0.2% CPB A and B의 5개 처리구를 4반복으로 배치하여 시행하였다. 기초사료는 사육전기에 3,169와 3,149kca./kg ME와 21% CP를 급여하였고, 사육후기에는 3,192와 3,172 kcal/kg ME와 19% CP사료를 급여하였다. 증체량, 사료섭취량, 사료요구율은 주간별로 5주간 측정하였으며 실험 종료시에 장내 혐기적인 미생물 수를 조사하였다. 실험 3에서 24주령 로만 갈색계 450수를 이용하여 처리구는 실험 2와 동일하게 8주간 시행하였다. 처리구당 반복은 18수씩 5반복으로 90수 전체 450수를 이용하였다. 사료는 대상에너지 2,720 kcal/kg와 CP는 18.5%로 하였다. 처리구당 산란율, 난중, 사료섭취량, 사료요구율, 계란의 품질을 주간 별로 조사하였으며, 실험종료시에 혈액의 성상을 조사하였다. 실험 1에서 PBSL 0.5%, W10 0.5%, CPB A, B 0.2% 처리구의 PDI는 95.9, 95.9, 95.8. and 95.7로서 대조구에 비하여 현저하게 높았다.(P<0.05). 그러므로 이러한 결과는 실험2와 3에 적용하였다. 실험 2에서 PBSL 처리구의 증체량은 대조두와 통계적인 차이는 없었지만 W10과 CPB A와는 현저하게 차이가 있었다. (P<0.05). 사료요구율은 PBSL이나 다른 사료결착제 처리구에서 대조구에 비하여 개선되는 경향을 보였다.

본 시험의 목적은 국내에서 분리된 강병원성 뉴캣슬병 바이러스를 SPF(specific pathogen free)닭에 접종을 하여 병리학적 변화와 각 조직내의 뉴캣슬병 바이러스 항원의 분포를 조사하여 국내분리 바이러스들간의 조직내 친화성을 비교함으로써 분리 바이러스간 병리학적 차이점을 규명하는 것이다. 병리학적 검사를 위하여 가능한 대부분의 조직을 채취하였으며 항원검사는 특이성을 높이기 위하여 단클론성 항체를 이용하였다. 검사결과를 요약하면 뉴캣슬병 바이러스를 접종한 모든 닭들은 접종 후 3일부터 폐사가 시작되어 접종 후 9일에 100% 폐사하였으며 대부분의 접종군은 접종후 5일에서 6일 사이에 폐사가 있었으나 8248 계군은 접종후 7일에서 9일 사이에 모두 폐사를 하여 다른 접종군과 확연히 다른 폐사양상을 보였다. 그러나, 폐사한 닭들은 접종 바이러스에 관계없이 임상증상은 비슷하였다. 일반적으로 접종 후 3일이 경과하면 급격한 활동저하와 함께 빈사상태를 보였고 일단 빈사상태까지 진행된 접종 닭은 다시 회복하지 못하였다. 병리조직학적 소견은 과거 연구자들이 강병원성 뉴캣슬병에서 흔히 관찰하였던 장, 기낭, 폐장에서의 출혈소견 들을 발견할 수 있었으며 접종한 바이러스에 따른 병리학적 변화의 차이는 뚜렷하지 않았다. 공통적인 병변으로서 대표적인 것은 면역기관인 F. bursa, 흉선, 비장에서의 탐식세포 및 단핵 세포의 괴사로서 과거의 문헌들에서 기술된 병변과 비교해 볼 때 이 면역기관에서의 괴사소견은 강병원성 뉴캣슬병의 가장 특징적인 조직학적 병변인 것으로 판단된다. 조직병변과 바이러스간의 유기적 관계를 알아보기 위해 면역화학조직검사법(Immuno histo chemistry)이 많이 이용되는데 본 실험에서도 이 방법을 사용하여 국내분리주들간의 조직친화성을 비교하여 보았다.지하였으며 폐사율은 각각 1.9%, 1.9%, 2.9%와 1.0%로써 대조군의 4.8%에 비하여 낮은 수준을 보여 질병에 대한 저항능력이 향상되었다.kg/h로 분석되었다. 6. 건조작업 중에 소요되는 전력은 9.5~19.3 Wh/kg이었으며, 연료효율 6.9~9.3kg-$H_2$O/L와 열효율 50.2~65.1%에 의한 수분증발량은 124.0~125.4 kg-$H_2$O/h 로 나타났다. 7. 건조기의 열효율을 70%를 기준으로 평가한다면, 시작기의 열효율은 조금 낮은 수준이며, 열관리 계통의 개선을 필요로 하였다. 8. 제작된 계분건조기의 시작기를 15,000수 규모의 양계농장에서 사용한다면 관행의 계분처리 방법에 비하여 약 590,700(원/월)의 비용을 절감할 수 있을 것으로 추정되었다.

The goal of this paper was to examine the qualify zygote-acquiring method for in-vitro culture and the in-vitro culture method of the acquired zygote from a technological perspective. We have reported the results on the introduction of foreign DNAs using the described culturing method. After performing in-vitro and surrogate eggshell culture on a zygote acquired from the abdomen of a hen, 25.8% hatchability was acquired. After microinjecting foreign DNAs into the acquired zygote and performing in-vitro and surrogate eggshell culture using the same method, 13.1∼11.7% hatchability was acquired. Having compared the developments of the control subjects and the experimental subjects, the viability of the experimental subjects on the 4∼5th day of culturing was much lower compared to that of the control subjects. This is a result that shows that the microinjection process of foreign DNAs might have a negative effect on the existence of the embryo; therefore, various technical attempts should be made to minimize such negative effects. Having microinjected foreign DNAs into the zygote of a hen to produce transgenic chickens, 3 transgenic founders were Produced and 70 G1 progeny were produced as a result of the progeny test that had been performed to the present.

A novel sterategy has been established to determine the origin of the Primordial Germ Cells (PGCs) in avian embryos directly and the developmental fate of the PGCs for the application to Poultry biotechnology. Cells were removed from 1) the centre of area pellucida, 2) the outer of area pellucida and 3) the area opaca of the stage X blastoderm (Eyal-Giladi & Kochav, 1976). When the cells were removed from the centre of area pellucida, the mean number of circulating PGCs in blood was significantly decreased in the embryo at stage 15 (Hamburger & Hamilton, 1951) as compared to intact embryos. When the cells were replenished with donor cells, no reduction in the PGCs number was observed. The removal of cells at the outer of area pellucida or at the area opaca had no effect on the number of PGCs. In case, another set of the manipulated embryos were cultured ex vivo to the hatching and reared to the sexual maturity, the absence of germ cells and degeneration of seminiferous tubules was observed in resulting chickens derived from the blastoderm in which the cells were removed from the centre of the area pellucida. It was concluded that the avian Primordial Germ cells are originated at the center of area pellucida. Developmental ability of the cells to differentiate into somatic cells and germ cells in chimeras were analyzed. Somatic chimerism was detected as black feather attributed from donor cells. Molecular identification by use of female - specific DNA was performed. It was confirmed that the donor cells could be differentiated into chimeric body and erythrocytes. Donor cells retained the ability to differentiate into germline in chimeric gonads. More than 70% of the generated chimeras transmitted donor derived gametes to their offspring indicating that the cells at the center of area pellucida had the high ability to differentiate into germ cells. A molecular technique to identify germline chimerism has been developed by use of gene scan analysis. Strain specific DNA fragments were amplified by the method. It would be greatly contributed for the detection of germline chimerism. Mixed- sex chimeras which contained both male and female cells were produced to investigate the developmental fate of male and female cells in ovary and testes. The sex combinations of donor and recipient of the resulting chimeras were following 4 pairs; (1) chimeras (ZZ/ZZ) produced by a male donor (ZZ) and a male recipient (ZZ), (2) chimeras (ZW/ZW) produced by a female donor (ZW) and a female recipient (ZW), (3) chimeras (ZZ/ZW) Produce by a male donor (ZZ) and a female recipient (ZW), (4) chimeras (ZW/ZZ) produced by a female donor (ZW) and a male recipient (ZZ). It was found that genetically male avian germ cells could differentiate into functional ova and that genetically female germ cells can differentiate into functional spermatozoa in the gonad of the mixed- sex chimeras. An ability for introduction of exogenous DNA into the PGCs from stage X blastoderms were analyzed. Two reporter genes, SV-$\beta$gal and RSV-GFP, were introduced into the PGCs. Expression of bacterial/gal was improved by complexing DNA with liposome detectedcc in 75% of embryos at 3 days embryos. At the embryos incubated for 1 day, expression of the GFP was observed all the embryos. At day 3 of incubation, GFP was detected in about 70% of the manipulated embryos. In case of GFP, expression of the transgene was detected in 30 %e of the manipulated embryos. These results suggested that the cells is one of the most promising vectors for transgenesis. The established strategy should be very powerfull for application to poultry biotechnology.

Using bioinformatic tools for searching the massive genome databases, it is possible to Identify new genes in few minutes for initial discoveries based on evolutionary conservation, domain homology, and tissue expression patterns, followed by further verification and characterization using the bench-top works. The development of high-density two-dimensional arrays has allowed the analysis of the expression of thousands of genes simultaneously in the humans, mice, rats, yeast, and bacteria to elucidate the genes and pathways involved in physiological processes. In addition, rapid and automated protein identification is being achieved by searching protein and nucleotide sequence databases directly with data generated from mass spectrometry. Recently, analysis at the bio-chemical level such as biochemical screening and metabolic profiling (Biochemical genomics) has been introduced as an additional approach for categorical assignment of gene function. To make advantage of recent achievements in computational approaches for facilitated gene discoveries in the avian model, chicken expression sequence tags (ESTs) have been reported and deposited in the international databases. By searching EST databases, a chicken heparanase gene was identified and functionally confirmed by subsequent experiments. Using combination of sub-tractive hybridization assay and Genbank database searches, a chicken heme -binding protein family (cSOUL/HBP) was isolated in the retina and pineal gland of domestic chicken and verified by Northern blot analysis. Microarrays have identified several host genes whose expression levels are elevated following infection of chicken embryo fibroblasts (CEF) with Marek's disease virus (MDV). The ongoing process of chicken genome projects and new discoveries and breakthroughs in genomics and proteomics will no doubt reveal new and exciting information and advances in the avian research.

Gene and cell transfer technique will serve as a powerful tool for the genetic improvement of the poultry and to yield useful products. For avian transgenesis, Japanese quail may serve as an excellent animal model because of its small body size and fast growth rate. Recent progress was described on the manipulation of quail embryos such as the introduction of foreign genes and cells, and the subsequent culturing of the manipulated embryos yielding hatchlings. Intraspecific donor-derived offspring have been available in quail, however, further investigation will be required to obtain interspecific offspring with the aim of rescuing endangered species. Trans genesis will also be useful for improving the profitability and quality of poultry stocks and for developing stocks with novel uses. Considerable progress should soon be made toward the production of transgenic poultry. The key feature of the procedure described here is that embryos are initially taken out from the shell for ease of manipulation and then placed back in culture in addition to various operations midway during culture.

Vaccination is one of the most important and cost-effective methods of preventing infectious diseases. Over the past decade, scientific in molecular biology and immunology have improved understanding of many diseases and led to the development of novel strategies for vaccination. An ideal vaccine would induce effective immunity specific for the type of infection, have long duration, require minimal or no boosters, have safety, would not induce adverse reaction, and be easy to administer. The desire to meet these criteria has resulted in the development of vaccines that do not depend on the use of the viable disease agent. It is not the intent of this review to give an extensive review of the field of vaccinology, but rather to address characteristics of conventional and genetically engineered vaccines.