禽类原始生殖细胞与转基因研究进展

综述了鸟类原始生殖细胞的特性，起源、迁移等以及对PGCs进行操作和培养，转基因鸡的研究现状和进展。     

鹅源禽痘病毒在鸡胚中的增殖及其理化特性鉴定

为填补国内外水禽源痘病毒培养特性和理化特性研究方面的空白,对国内检测并鉴定的鹅源禽痘病毒,用鸡胚进行分离培养和病毒理化特性研究。将鹅皮肤痘疹样本研磨后,接种SPF鸡胚进行连续传代培养。病理检查和PCR检测证实,病毒可在鸡胚中增殖,且经连续5轮传代后,鸡胚病变及病毒滴度趋于稳定。鸡胚中可以观察到绒毛尿囊膜水肿增厚及白色痘斑等禽痘特征性病变。对采集病变的绒毛尿囊膜进行电子显微镜观察,可见病毒包涵体和典型禽痘病毒粒子。利用建立的鸡胚培养体系,对该病毒理化特性进行鉴定,发现病毒对热(55℃)、酸(pH3)、碱(pH11)、胰蛋白酶、乙醚和氯仿敏感。本研究首次建立了鹅源禽痘病毒鸡胚培养体系并对其理化特性进行了鉴定,从而为系统开展该病毒生物学特性和防控技术研究提供了必要的技术基础。     

1例鸡白痢沙门氏菌的分离鉴定与耐药性分析

为查明河南省新乡市某规模化蛋鸡场疑似沙门氏菌感染病例的病原，进而制定合理的治疗方案，本研究无菌采集疑似病雏鸡肠管样品，用革兰氏染色、培养特性观察、生化试验、PCR方法对其进行确诊，并进行病原回归试验，采用药敏纸片琼脂扩散法（K-B法）分析分离菌对19种常见抗生素的耐药性。结果显示，本试验成功分离了一株革兰氏阴性短杆菌，该分离菌符合鸡白痢沙门氏菌的培养特性和生化特性；PCR成功扩出invA基因，通过与GenBank数据库比对分析确定该细菌为鸡白痢沙门氏菌；用分离细菌株感染SPF鸡能复制出与自然感染一致的病例，说明鸡白痢沙门氏菌是造成本养鸡场雏鸡发病的主要病原；分离株具有较强的致病性和多重耐药性，对阿米卡星、苯唑青霉素、青霉素耐药，对复达欣、氨苄青霉素、头孢曲松等药物敏感，将敏感抗生素用于临床治疗效果明显。结合以上结果，本研究提出该病的具体防治措施，并取得了较好的效果，为鸡白痢沙门氏菌的分离鉴定及防治提供了参考，对鸡白痢沙门氏菌病病原的早期诊断和治疗具有重要临床意义。     

禽类原始生殖细胞与转基因研究进展

综述了鸟类原始生殖细胞的特性、起源、迁移等以及对 PGCs进行操作和培养 ,转基因鸡的研究现状和进展     

短乳杆菌的分离鉴定及其对鸡肠粘膜SIgA分泌的影响

用RS培养基，从SPF鸡的消化道分离到1株乳酸杆菌。该菌生长速度快，耐酸，耐胆汗，对肠道粘膜有较强的粘附力，能提高鸡肠粘膜的SIgA滴度。对该菌的形态特征、培养特性、生理生化特性、药物的敏感性及G C等进行试验，证明该菌为短乳杆菌。该菌可作为禽用闪生素的候选菌株。     

鸡球虫体外培养模型及应用研究进展

鸡球虫病是由艾美耳球虫寄生于鸡肠上皮细胞内所引起的一种寄生性原虫病，感染鸡体重减轻、营养不良、肠道损伤等，严重危害养鸡业的健康发展。球虫的生命周期包括从无性阶段到有性阶段的转换并局限于一个宿主。目前对球虫的研究主要包括在细胞生物学及其不同生命阶段的蛋白质表达和运输机制、宿主细胞入侵和宿主-寄生虫相互作用、新型药物靶标筛选等方面。鉴于研究问题的多样性，以及减少和取代动物试验的要求，建立高效的体外培养模型成为研究鸡球虫生物学特性、抗球虫疫苗和药物的基本条件。笔者重点阐述了鸡球虫体外培养模型及其在药物抗虫效果研究中的应用，包括鸡胚培养模型、二维培养模型(原代细胞培养模型、传代细胞系培养模型)及三维类器官培养模型。越来越多培养技术的突破为研究球虫生命周期阶段和干预策略开辟了新的途径，也为深入了解鸡球虫致病机制、抗球虫药物研发提供依据。     

鸡传染性喉气管炎及其防制研究

作者论述了鸡传染性喉气管炎的病原及培养特性、发病机制、诊断方法及其防制，并对传染性喉气管炎的控制和根除进行了展望。     

MHC及其在肉鸡和蛋鸡中的不同

本文综述了鸡HHC的结构、组成等特性，并介绍了其在肉鸡和蛋鸡中的不同。最后，总结了鸡HHC的作用．并对其在鸡育种中的应用前景进行了展望。     

鸡大肠杆菌菌种选育及冻干藏试验

为试验鸡大肠杆菌病中草药佐剂多价灭活疫苗而进行了制造和检验用菌种的选育试验。经染色特性、培养特性、生化特性及血清学特性等的观察和测定，选用Ａ、Ｂ、Ｃ、ＳＥ４株血清型分别是Ｏ１、Ｏ２、Ｏ７８及Ｏ２４的鸡Ｅ．ｃｏｌｉ作为标准菌株，现地分离和鉴定有代表性疫区或病鸡群病原血清型作为地方菌株，经毒力和免疫原性测定，结果证实，该４株血清型大肠杆菌作为疫苗制造和检验用菌种较好，并分别进行了冷冻真空干燥保存试验，     

鸡胚胎体外操作技术的研究现状和进展

本文介绍了禽类胚胎操作技术方面的概况，对受精卵收集及体外受精、早期胚胎体外培养、人造材料蛋壳培养胚胎、代用蛋壳培养胚胎、嵌合体鸡的生产、转基因鸡生产等的进展及发展进行了综述。     

A comparison of routine culture with polymerase chain reaction technology for the detection of Mycoplasma species in feline nasal samples.

Nasal flush samples were collected from 20 cats and submitted for Mycoplasma culture and polymerase chain reaction (PCR). Nasal biopsy samples were also obtained from each cat and simultaneously evaluated for Mycoplasma by standard culture and PCR. Concordance of the test results was determined through calculation of the kappa statistic. In 6 cats, nasal flush samples were culture positive for Mycoplasma. PCR was positive in each culture-positive cat and also positive in 1 flush sample that was culture negative. DNA sequencing of the PCR product from the culture negative flush sample identified the organism as Mycoplasma arginini. All other flush samples that were culture negative were also PCR negative (kappa = 0.89). Nasal biopsy samples from 7 cats were culture positive for Mycoplasma, and all were PCR positive. Biopsy samples that were culture negative for Mycoplasma were also PCR negative (kappa = 1.0). Results of culture and PCR for both nasal flush and biopsy were concordant in 19 of 20 cats, and PCR was able to identify an unusual Mycoplasma species that did not grow in culture. In most cats, organisms could be detected in either nasal flush or biopsy samples. In this study, PCR provided rapid and sensitive detection of Mycoplasma species in nasal samples from cats and detected 1 organism that did not grow in culture.     

Evaluation of highly sensitive indigenous milk ELISA kit with fecal culture, milk culture and fecal-PCR for the diagnosis of bovine Johne's disease (BJD) in India

Country lacks indigenous diagnostic kits against Johne's disease in animals. Indigenous ELISA and IS 900 PCR kits, originally developed for goats and sheep, have been adapted for screening of lactating cows. Multiple diagnostic tests were used to screen 26 lactating dairy cows against Johne's disease. Milk ELISA was evaluated with fecal culture, milk culture and fecal PCR. Of the 26 samples from lactating cows, 84.6, 96.1, 88.4 and 23.0% were positive in fecal culture, milk culture, m-ELISA and m-PCR, respectively. Comparatively milk sediment and milk fat culture detected 84.6 and 76.9% cows positive, respectively. Comparatively fecal culture and milk culture detected 84.6 and 96.1% cows positive, respectively. M-ELISA detected 11.5, 0.0, 11.5, 61.0 and 15.3%, cows as negative, suspected, low positive, positive and strong positive, respectively. There was good correlation between milk and fecal culture with m-ELISA. Three negative cows in m-ELISA were also detected in milk and fecal culture. Of the 26 decontaminated fecal samples, 23.0% cows were positive using specific IS 900 f-PCR. Comparative evaluation of m-ELISA with fecal and milk culture showed agreement in 80.7 and 84.6% cows, respectively. Sensitivity of m-ELISA with respect to fecal and culture was 90.9 and 95.6%, respectively. Comparative evaluation of four tests (milk culture, fecal culture, m-ELISA and f-PCR) showed that only 15.3% cows were detected in all the four tests. In three tests (fecal and milk culture and m-ELISA), 57.6% cows were detected positive. None of the cow was exclusively detected in f-PCR. Of the four diagnostic tests used milk culture was most sensitive (96.15%), followed by fecal culture (86.6%), m-ELISA (76.9%) and IS 900 PCR (23.0%) for the diagnosis of bovine Johne's disease (BJD). Milk ELISA detected only one cow extra, which was negative in milk culture. In view of the simplicity, rapidity and efficacy present milk ELISA kit employing soluble protoplasmic antigen from native Map 'Bison type' genotype of goat origin can be reliable for screening of bovine population against Johne's disease in India.     

Detection of Mycobacterium avium subsp. paratuberculosis in milk from clinically affected cows by PCR and culture

Milk and faeces samples from cows with clinical symptoms of paratuberculosis were examined for the presence of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) by culture and PCR. M. paratuberculosis was cultivated in variable numbers from faeces or intestinal mucosa in eight of 11 animals. In milk from five cows (all faeces culture positive), we cultivated a few colonies of M. paratuberculosis (<100 CFU per ml). Milk samples from two cows were PCR positive (both animals were faeces culture positive, and one cow was milk culture positive). One cow was culture negative on intestinal mucosa, but culture positive in milk, and two cows were negative in culture and PCR from both faeces and milk. In conclusion, the presence of M. paratuberculosis could be detected in raw milk by PCR, but cultivation of milk was more sensitive.     

体外培养鸽嗉囊组织形态学、酶活力及基因表达动态变化规律

本试验旨在探究体外培养鸽嗉囊组织形态学、酶活力及基因表达变化规律。取60周龄美国王鸽嗉囊组织于体外培养7 d,动态检测其形态学、代谢和凋亡相关酶活力以及角蛋白和凋亡相关基因表达的变化。形态学结果表明:鸽嗉囊组织于体外培养第2~4天结构清晰,上皮层细胞排列紧密,体外培养5 d后嗉囊结构显著退化,形态学受到破坏。酶活力和基因表达结果显示:Caspase-3酶活力于培养第1、5、6、7天显著高于培养第2~4天(P0.05);琥珀酸脱氢酶、Na~+-K~+-ATP酶、Ca~(2+)-Mg~(2+)-ATP酶和总ATP酶活力以及Bcl-2和角蛋白19基因表达均于培养第1、6、7天显著下调(P0.05),于培养第2~4天显著升高(P0.05);Bak1基因表达于培养第2~5天稳定,随后于培养第6天显著上调(P0.05)。综上所述,体外培养鸽嗉囊组织形态学、酶活力和基因表达随培养时间延长而显著改变。本试验条件下,鸽嗉囊组织体外培养稳定时间为4 d,且开展后续药理、病理及生理学试验前的1 d预培养十分必要。     

动物早期胚胎体外培养的研究进展

胚胎的体外培养是动物胚胎工程技术的基础环节之一,也是胚胎移植重要的前期基础。由于在体外培养过程中,早期胚胎对环境的变化较为敏感,导致早期胚胎发育到囊胚的成功率不高,因此,如何提高囊胚率是核心问题。就动物早期胚胎体外培养的培养环境、胚胎培养液添加物以及胚胎体外培养体系等方面的研究进展进行综述,同时对胚胎体外培养技术目前存在的一些问题进行了总结,并对其前景做了展望。     

Microbiological and histopathological features of canine acral lick dermatitis

The purpose of this study was to investigate microbiological and histopathological features of canine acral lick dermatitis (ALD). Microbial characteristics of ALD are poorly described in current literature. If infection is recognized, antimicrobial selection is usually empirical, based on appearance, cytology or surface culture, rather than deep tissue culture. It was hypothesized that cultures obtained from deep tissue would yield different results than predicted by surface culture and cytology, and that isolates from ALD have unpredictable susceptibility patterns showing resistance to antibiotics routinely administered for canine pyoderma. Biopsies were obtained from 31 lesions and submitted for aerobic, anaerobic and fungal culture, and histopathological evaluation. Surface aerobic culture and susceptibility and cytology were obtained for comparison in 22 dogs. Skin scrapings and dermatophyte culture were performed. Bacteria were isolated in 30 of 31 cases. Staphylococcus intermedius was isolated in 58% of deep cultures. Twenty per cent of deep isolates were methicillin-resistant Staphylococcus species. Forty-eight per cent of cases yielded organisms defined as multidrug resistant on deep culture. Only 57% and 55% of bacteria isolated from tissue culture were sensitive to amoxicillin-clavulanic acid and cefazolin, respectively. Cytology and superficial cultures did not correlate well with deep cultures. Surface culture predicted deep tissue isolates in eight of 22 cases. Microsporum gypseum was isolated from one dog. Histopathological features included acanthosis, follicular elongation, lymphoplasmacytic dermal inflammation, folliculitis, furunculosis, perihidradenitis, hidradenitis and vertical streaking fibrosis. Lesions associated with ALD warrant tissue bacterial cultures as the majority of cases yielded positive growth of bacteria differing from superficial culture and often resistant to empirical drugs.     

马丁干粉培养基的性能研究

将研制的马丁干粉培养基与新鲜马丁培养基比较,证明用干粉培养基培养链球菌、多杀性巴氏杆菌和丹毒杆菌,其每1 mL中活菌数和灵敏度均与新鲜马丁培养基相当.干粉培养基用于生物制品的生产及检验更方便且质量稳定,可推广应用.     

悬浮培养工艺与转瓶培养工艺的比较分析

采用反应器全悬浮培养BHK21细胞生产口蹄疫病毒与微载体悬浮培养Vero细胞生产狂犬病毒分别与相应的转瓶培养工艺生产案例对比分析,比较悬浮培养工艺与转瓶培养工艺的生产效益。分析显示,与转瓶培养工艺相比,反应器悬浮培养工艺获得的细胞密度、病毒效价、产品的产量和质量明显提高,生产时的能耗和劳动力需求明显降低。结果表明悬浮培养工艺的生产效益明显高于转瓶培养工艺,适宜于国内生物制品工业化生产的升级换代。     

Culture of strategically pooled bovine fecal samples as a method to screen herds for paratuberculosis.

Fecal samples from 733 cows in 11 dairy herds with a low prevalence of paratuberculosis were cultured for the presence of Mycobacterium avium subsp. paratuberculosis both individually and after combining (pooling) in groups of 5. The culture procedure was the modified Jorgensen method, which uses NaOH and oxalic acid for decontamination and modified Lowenstein-Jensen agar slants for cultivation. Pooling was performed by mixing fecal samples from 5 animals ordered by age, herein referred to as strategic pooling. Culture of individual fecal samples detected M. a. paratuberculosis infections in 43 of the 733 cows and 7 of 11 infected herds (herd sensitivity = 64%). Culture of pooled fecal samples detected M. a. paratuberculosis in 28 of 151 pooled samples representing 8 of the infected 11 herds (herd sensitivity = 73%). Feces of the 43 culture-positive cows was included in 32 pools: of these 32 pools, 26 were culture positive and 6 were culture negative. In addition to the 26 positive pools containing feces from cows that were found culture positive on individual fecal samples, another 2 pools were culture positive, although comprised of feces from cows with negative results after culture of individual fecal samples. From the total of 45 infected cows that were found (43 by individual fecal culture and an additional 2 by pooled fecal culture), individual fecal culture detected 43 of these 45 (96%), while pooled fecal culture detected 39 (87%). Culture of strategically pooled fecal samples using the modified Jorgensen method was equivalent in herd sensitivity to the culture of individual fecal samples and is significantly less expensive.     

Comparison of culture, peroxidase-antiperoxidase reaction, and serum latex agglutination methods for diagnosis of chlamydiosis in pet birds

Comparison was made among results of cloacal specimen culture, and cloacal swab specimen (cytologic) peroxidase-antiperoxidase (PAP), serum latex agglutination (LA), and tissue PAP assays for diagnosis of chlamydiosis in 144 birds. Swab specimen PAP findings correlated poorly with LA results and failed to predict the LA test result in any bird. Only 1 cloacal swab specimen was regarded as PAP-positive and was from the cloaca of a bird from which chlamydiae were isolated in culture. The sensitivity of swab specimen PAP, compared with culture results, was 33%, whereas specificity was 94%. In this study, swab specimen PAP was a less sensitive test, compared with culture, than was reported in a previous study. The sensitivity of LA in identifying birds that were cloacal culture-positive was poor; true-positive results were not detected, compared with culture results. The specificity of the LA method was 93%, compared with culture results. Results of the tissue PAP method correlated with culture results in the 3 birds for which both tests were performed.