This study was carried out to investigate the antimicrobial resistance pattern and distribution of resistance gene determinants in fecal E. coli from chicken. Antimicrobial susceptibility test was performed on a total of 109 fecal E. coli isolates from chicken, collected in Gyeonggi, Chungbuk, Jeonnam and Jeonbuk province from March to November 2003, by the disk agar diffusion method. Eighteen commonly used antimicrobial agents approved in Korea as veterinary medicine were tested: ampicillin (AM), amoxicillin/clavulanic acid (AMC), cephalothin (CF), cefozolin (CZ), cefoxitin (FOX), cefotaxime (CTX), cefepime (FEP), imipenem (IPM), streptomycin (S), gentamicin (GM), amikacin (AN), ciprofloxacin (CIP), enrofloxacin (ENO), norfloxacin (NOR), trimethoprim/sulfamethoxazole (SXT), erythromycin (E), chloramphenicol (C) and tetracycline (TE). Higher resistance rates (≥50%) were observed against 9 antimicrobial agents including AM, CF, S, CIP, ENO, NOR, SXT, E and TE. Resistance was most frequent for TE in 105 E. coli isolates (96.3%). Twenty-two isolates (20.2%) of the isolates showed multiple antimicrobial resistance to 8, and 19 isolates (17.4%) showed to 7 antimicrobial agents. The distribution of the resistance gene determinants for S and TE was assessed by PCR in resistant isolates. Thirty isolates possessed the strA, strB, and aadA gene, 25 isolates possessed the strA and strB gene among the 66 streptomycin-resistant isolates. Fifty one isolates possessed only the tetA gene, 22 isolates possessed the tetA and tetB gene, 11 isolates possessed only the tetB gene among the tetracycline-resistant isolates.

The purpose of this study is to establish a basic culture system enabling in vitro culture of chicken blastodermal cells and to test the feasibility of retrovirus-mediated gene transfer to the cultured cells. The blastodermal cells were isolated from freshly laid eggs of stage X and cultured with or without STO feeder layer cells. Stem cell-like morphology was maintained after multiple passages and RT-PCR analysis proved expression of several stem cell specific genes. Immunocytochemical analysis using antibodies of anti-EMA-1 and anti-SSEA-1 also showed the feature of stem cells. Infection of the cultured blastodermal cells with LNCGW retrovirus vector resulted in successful transfer of foreign genes. The results of this study may be useful in establishing stem cell-mediated transgenic chicken production.

We report here the production of transgenic chickens that can regulate human erythropoietin (hEPO) gene expression. The glycoprotein hormone hEPO is an essential for viability and growth of the erythrocytic progenitors. Retrovirus vector system used in this study has two features including tetracycline-controllable promoter and woodchuck hepatitis virus posttranscriptional regulator element (WPRE). The former is for to reduce the possibility of physiological disturbance due to constitutional and unregulated expression of hEPO gene in the transgenic chicken. The latter is for maximum expression of the foreign gene when we turn-on the gene expression. A replication-defective Moloney murine leukemia virus (MoMLV)-based vectors packaged with vesicular stomatitis virus G glycoprotein (VSV-G) was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 325 injected eggs, 28 chicks hatched after 21 days of incubation and 16 hatched chicks were found to express the hEPO gene delivered by the vector. The biological activity of the recombinant hEPO in transgenic chicken serum was comparable to its commercially available counterpart. The recombinant hEPO in transgenic chicken serum had N- and O-linked carbohydrate simillar to that produced from in vitro cultured cells transformed with hEPO gene.

The acaricidal activities of Illicium verum fruit-derived materials against adults of Dermanyssus gallinae were examined using the direct contact application method. Based on laboratory tests, an acaricidal constituent of I. verum fruit was determined because of its potent activity. Results were compared with those of the currently used acaricides such as dichlorvos, diazinon, and carbaryl. The acaricidal principle of I. verum fruit was identified as (E)-anethole using a GC-MS. Its acaricidal activity was compared with those of 12 compounds having a similar chemical moiety. Based on the LD50 values, the acaricidal activities of (+)-or-(–)-neomenthol were the strongest (0.01 ㎎/㎠]) and (E)-anethole, (+)-or-(–)-menthol, (±)-isoborneol, (–)-menthone, and (1S)-endo-(–)-borneol showed similar results (0.02 ㎎/㎠), and (1R)-(+)-camphor and (+)-menthone also gave good activities (0.03 and 0.04 ㎎/㎠, respectively). These compounds showed more toxic acaricidal activities than diazinon and carbaryl, 0.05 and > 0.2 ㎎/㎠, respectively, but were not comparable to that of dichlorvos with 0.0002 ㎎/㎠. These results indicate that the I. verum fruit-derived materials and tested compounds descried as poultry red mites-control agents could be useful for managing field populations of D. gallinae.

본 연구는 vesicular stomatitis virus G glycoprotein (VSV-G)으로 피막이 형성되는 replication-defective MoMLV-based vector를 이용한 hTPO 헝질전환 닭의 생산에 관한 연구이다. 실험에 사용한 retrovirus vector의 구조는 hTPO 유전자의 발현 조절을 위해 internal promoter인 hCMV promoter를 이용하였으며 외래 유전자의 발현을 증가시키기 위해 woodchurk hepatitis virus posttranascriptional regulatory element (WPRE) 서열을 도입하였다. 재조합한 vector는 GP2 293 포장세포에 도입하여 virus를 생산하였으며 이 virus를 이용하여 감염시킨 여러 표적세포에서 hTPO의 발현과 생물학적 활성을 확인하였다. 재조합 hTPO의 생물학적 활성은 시판되고 있는 재조합 hTPO에 비해 우월한 것으로 확인되었다. hTPO 형질전환 닭의 생산을 위하여 1,000배 이상 고농도로 농축된 virus를 stage X 단계의 계란의 배반엽 층에 미세주입하여 대리난각 방법으로 배양하였다. 미세주입한 132개의 계란 중 21일 후에 11개의 계란에서 병아리가 부화하였으며 그중 4마리가 형질전환 개체로 확인되었다. 그러나 생산된 4마리 중 3마리가 부화 후 1개월 이내에 원인불명으로 사망하였다. 본 연구의 의의는 상업적 이용 가능성이 있는 생물학적 활성을 가진 사람의 cytokine 단백질의 대량 생산을 위한 생체 반응기로서의 형질전환 닭 개발의 시례를 제공하는데 있다.