鸡免疫器官中血管活性肠肽的分布特点

应用免疫组化超敏SP法对鸡不同生长阶段免疫器官中血管活性肠肽(VIP)的表达特点进行了研究,结果显示,胸腺中VIP阳性反应主要分布于被膜及髓质,而皮质内反应较弱;随日龄增加,VIP在被膜和髓质内一直维持较强的表达;实质中VIP强阳性淋巴细胞数量随日龄逐渐增多,至42 d开始减少.脾脏中VIP阳性反应主要出现于被膜及红髓,并在整个生长阶段维持较强的表达,红髓的脾索和脾窦间还有较多的巨噬细胞呈VIP强阳性反应;白髓中仅见动脉周围淋巴鞘中央动脉有VIP的表达,并随日龄的增加逐渐减弱.法氏囊中VIP的表达范围广泛,涉及黏膜、黏膜下层和肌层,尤以黏膜上皮细胞和法氏囊小结髓质的表达为典型;随着日龄增加,黏膜上皮细胞VIP表达逐渐减弱,囊小结髓质一直保持较强的VIP阳性着色,囊小结皮髓质交界处上皮细胞层内VIP强阳性细胞有逐渐增多的趋势.鸡免疫器官中VIP阳性反应物质的广泛分布,提示这3种器官是神经系统以外产生VIP的重要部位,法氏囊中的VIP可参与鸡免疫器官的功能调节,它可作为一种信号分子介导神经内分泌系统和免疫系统之间的相互作用.     

鹅副粘病毒的组织嗜性

用单克隆抗体介导的免疫过氧化物酶（MC－IP）技术，对21只鹅副粘病毒病鹅体内的病毒抗原进行定位，结合组织病理学观察结果，探讨了鹅副粘病毒的组织嗜性。结果显示，除脑和心脏未检测到病毒外，在气管、肺、食道、肝脏、腺胃、胰腺、肠、哈氏腺、胸腺、脾脏、法氏囊、肾脏等器官都能检测到病毒，病毒抗原定位于上述器官的各种上皮细胞，淋巴细胞、网状细胞和巨噬细胞的胞浆内，其中以胃肠粘膜上皮细胞和法氏囊、胸腺、脾脏的淋巴细胞和网状细胞内检出率高，阳性反应强。这些结果提示，鹅副粘病毒是一种泛嗜性病毒，胃肠粘膜上皮和淋巴组织是其主要侵嗜部位。     

鸡肾型传染性支气管炎病毒组织嗜性的研究

本研究用鸡肾型传染性支气管炎病毒 (肾型 IBV) C90 0 1株人工感染 14日龄 SPF鸡 ,于接种后定期采集病鸡的各组织器官制备成石蜡切片 ,用建立的检测石蜡切片中肾型 IB病毒的免疫酶组化染色技术 ,对人工感染肾型传染性支气管炎 (肾型 IB)鸡发病后病毒的组织器官亲嗜性和动态分布规律进行了研究。结果表明 ,肾型 IBV在胞浆内复制 ,主要亲嗜气管黏膜的上皮细胞和固有层腺体细胞、肺各级支气管上皮细胞以及肺房和呼吸性毛细管上皮细胞、气囊上皮细胞、肾和输尿管上皮细胞、消化道上皮和固有层腺体细胞、肝小叶间胆管上皮细胞、胰腺导管上皮细胞和腺泡上皮细胞、法氏囊淋巴滤泡髓质区淋巴细胞和网状细胞、胸腺小叶髓质区淋巴细胞和网状细胞、脾小体淋巴细胞、盲肠扁桃体弥散性淋巴组织的淋巴细胞以及心肌细胞 ,病毒出现的先后顺序为气管、肺脏、肾脏、输尿管 ,消化道、法氏囊、肝脏、胰脏、胸腺和气囊 ,盲肠扁桃体 ,脾脏 ,心肌。病毒在肾脏和输尿管持续 2 0天 ,气管为 13天 ,消化道和法氏囊为 11天 ,肝脏、胰脏和肺为 10天 ,胸腺为 6天 ,气囊为 5天 ,盲肠扁桃体、脾脏、心肌呈一过性感染     

Ghrelin在BALB/c小鼠胸腺中的定位分布与发育性表达

本试验应用免疫组织化学方法和显微图像分析技术,探讨了Ghrelin在1、3、5、6、12月龄BALB/c小鼠胸腺中的定位分布和发育性表达规律。结果表明,Ghrelin免疫阳性反应主要分布于胸腺髓质内,而胸腺皮质中较少。Ghrelin阳性细胞类型包括被膜下胸腺上皮细胞、星形胸腺上皮细胞、髓质胸腺上皮细胞、胸腺小体上皮细胞、巨噬细胞、胸腺髓质的部分淋巴细胞,可能还有胸腺树突状细胞。Ghrelin在小鼠胸腺中的表达量,1、3月龄之间差异不显著(P0.05),5、6月龄之间差异也不显著(P0.05),但5、6月龄显著低于1、3月龄(P0.05),而12月龄又显著低于5、6月龄(P0.05)和极显著低于1、3月龄(P0.01),即随着年龄的增长,胸腺中的Ghrelin表达量逐渐减少。结果提示,胸腺的年龄性退化可能与Ghrelin的表达量下降有关。     

家禽肠道黏膜的作用及保护

家禽肠道黏膜由黏膜上层、固有层和黏膜肌层构成。黏膜上皮为单层柱状上皮,上皮细胞之间分布有许多杯状细胞,还有银亲合细胞,特别是十二指肠前段,这种细胞更多。固有膜由含有较多细胞成分的结缔组织构成,其中有血管、神经和肠腺,有时还有弥散性淋巴组织,在局部     

传染性支气管炎弱毒苗黏膜途径免疫对雏鸡气管及哈德氏腺SIgA细胞分布的影响

采用传染性支气管炎弱毒苗对1日龄雏鸡分别进行点眼和滴鼻免疫.取气管和哈德氏腺做石蜡切片,以葡萄球菌A蛋白-辣根过氧化酶(SPA-HRP)免疫组化染色,对气管和哈德氏腺中SIgA细胞的分布、数量变化进行了观察.结果表明:SIgA阳性细胞分布在哈德氏腺的间质中;气管中的SIgA细胞主要分布在黏膜上皮细胞之间和固有层中,局部黏膜上皮形成的隐窝内也分布有SIgA细胞.数量统计分析表明:免疫组的哈德氏腺和气管中SIgA细胞数量显著高于对照组;点眼免疫的SIgA细胞数量比滴鼻免疫的高;加强免疫则使SIgA细胞数量进一步增多.     

1株H5N1亚型高致病力禽流感病毒人工感染鸭的抗原定位观察

通过对鸭静脉接种或眼-鼻-口腔-泄殖腔接种禽流感病毒A/duck/Guangdong/220/2004(H5N1)后,采用免疫组织化学方法检查病毒核蛋白抗原,探讨了该病毒在组织器官中的定位与分布.结果表明,病毒核蛋白抗原主要存在于鸭的心肌细胞、肝细胞、枯否氏细胞、胰腺泡上皮细胞、肾小管上皮细胞、血管内皮细胞和血液吞噬性自细胞;在气管和支气管黏膜上皮,脑组织,肠黏膜上皮,胸腺、法氏囊、脾脏和肺脏的巨噬细胞或组织细胞以及坏死的细胞碎片中也可检查到少量病毒核蛋白抗原;而在食管、胃、睾丸、卵巢、甲状腺、肾上腺、哈德氏腺、骨骼肌等未检查到该抗原.     

鸡CDC25B-mRNA探针制备及其在鸡十二指肠黏膜的原位杂交反应

应用RT-PCR方法从鸡胚中扩增CDC25B基因片段，将其连接于pGM-Teasy。分别利用pGM-Teasy中T7和SP6启动子及其RNA聚合酶，以线性化的CDC25B/pGM-Teasy为模板转录合成正、反义DIG标记RNA探针。以合成的地高辛（DIG）标记的CDC25B-mRNA为探针，进一步应用原位杂交组织化学方法（in situhybridiza-tion histochemistry，ISHH）探查CDC25B-mRNA在鸡十二指肠黏膜的分布。结果表明，成年鸡十二指肠肠腺上皮细胞中有丰富的CDC25B-mRNA的转录，其中CDC25B-mRNA探针杂交阳性细胞在肠腺基底部和小肠绒毛中部分别占上皮细胞总数的81.60%±9.63%和36.21%±8.81%。原位杂交阳性产物大部分分布于十二指肠肠腺上皮“干细胞部”和“增生部”细胞的胞质和胞核，肠腺基底部杂交信号由下至上逐渐减弱，至小肠绒毛下部消失，转为阴性。在黏膜肌层以及肌肉层杂交反应呈阴性。本研究从基因水平证明了鸡十二指肠黏膜肠腺上皮“干细胞部”和“增生部”细胞中有CDC25B-mRNA的存在，表明该区域有活跃的增殖过程，以补充绒毛上端死亡脱落的细胞。     

宁都黄鸡小肠显微结构和甲苯胺蓝染色阳性细胞分布特点

通过石蜡切片和HE染色方法观察了宁都黄鸡小肠的显微结构,同时通过改良甲苯胺蓝染色的方法鉴定小肠中的甲苯胺蓝染色阳性细胞.结果显示,宁都黄鸡小肠与一般禽类小肠显微结构类似,黏膜下层缺乏十二指肠腺,黏膜固有层缺乏淋巴小结.小肠从前到后,杯状细胞逐渐增多.3段小肠的肠腺和回肠杯状细胞呈甲苯胺蓝染色阳性.小肠壁四层结构中都有肥大细胞的分布,主要分布于黏膜上皮细胞下、黏膜下层、肌层和外膜结缔组织中.肥大细胞体积较大,多数呈卵圆形,有的细胞形态不规则,甚至有突起.胞核染色呈深蓝色,胞质中含有紫红色颗粒.结果表明,宁都黄鸡小肠腺细胞和杯状细胞甲苯胺蓝染色呈阳性,说明其能够分泌有助于吸收的活性物质;小肠中具有丰富的肥大细胞,说明宁都黄鸡有较完善的肠道免疫屏障.     

贝氏莫尼茨绦虫感染绵羊对小肠局部黏膜淋巴组织的影响

为了研究贝氏莫尼茨绦虫自然感染绵羊对小肠黏膜免疫组织的影响,分别从宏观、微观及亚微观水平对自然感染贝氏莫尼茨绦虫的成年绵羊(感染组)肠道进行了细致地观察,并与正常成年绵羊(正常组)进行了比较.结果显示,感染组肠道所见虫体平均长度为1.5m,头节主要吸附在空肠淋巴集结分布丰富的部位,一般寄生数量为1～2条.眼观,虫体寄生部位黏膜增厚,表面有大量灰白色黏液附着,其间可见点状出血.镜下,局部黏膜上皮脱落,而在完整的黏膜上皮处,其上皮细胞、上皮内淋巴细胞、杯状细胞的数量都明显增多;固有层内毛细血管充血,淋巴细胞、浆细胞、弥散淋巴组织以及肠腺杯状细胞均有不同程度的增生,头节寄生处部分肠腺坏死;黏膜下层淋巴小结、淋巴集结显著增生,部分增生凸入固有层形成新的圆顶区;固有层与黏膜下层以及黏膜肌层可见大量嗜酸性粒细胞浸润.扫描电镜下,感染组肠黏膜上皮脱落;贝氏莫尼茨绦虫头节呈椭球状,有4个吸盘,无顶突,小沟,表面覆盖一层致密的微绒毛.研究结果表明,肠黏膜增厚,主要是局部黏膜免疫相关细胞在寄生虫虫体表面覆盖的微绒毛的不断刺激下,机体抗感染自身组织增生所致.成年绵羊对抗贝氏莫尼茨绦虫的感染可能是通过黏膜免疫相关组织增生来加强局部免疫力而实现的.     

Ghrelin阳性细胞在食蟹猴消化系统中的分布定位

目的:研究ghrelin阳性细胞在食蟹猴消化系统中的分布定位。方法:采用免疫组化SABC法检测ghrelin免疫阳性细胞在食蟹猴消化系统中的分布部位、含量及细胞形态。结果:ghrelin免疫阳性细胞分布于食蟹猴的胃、十二指肠、空肠前段、空肠中段以及胰腺中,而在口腔黏膜、食管、空肠后端至直肠、肝脏中均无表达。结论:食蟹猴内ghrelin阳性细胞的分布与人和其他动物的分布基本相似,ghrelin在进化上具有保守性,物种间存在差异性。     

禁食对生长期北京鸭腺胃ghrelin免疫阳性细胞的影响

本研究旨在观察禁食是否对生长期北京鸭腺胃ghrelin免疫阳性细胞生成有影响。用免疫组织化学的方法检测25、35、50日龄的北京鸭(已经禁食72 h)单位面积(mm2)腺胃组织中ghrelin阳性细胞数目。研究发现在这3个时期,不管是禁食组还是自由采食组,大量ghrelin阳性细胞多分布于腺胃深层复管状腺中;禁食组腺胃中gh-relin阳性细胞数目比自由采食组极显著增加(P0.01)。结果提示:禁食是影响腺胃产生有活性ghrelin的重要因素。     

Adenoviral gizzard erosion in commercial broiler chickens

Pathologic and immunohistochemical changes caused by group I of the fowl adenovirus (FAV) serotype-1 99ZH strain, isolated from broiler chickens exhibiting gizzard erosion, were investigated in commercial broiler chickens. One hundred twenty-two chickens were inoculated with the strain by both oral and ocular routes at 1, 3, or 5 weeks of age and euthanatized for necropsy within 4-18 days of inoculation. Focal gizzard erosions were observed in the inoculated chickens of each age group. A histologically degenerative koilin layer, necrotic mucosa, intranuclear inclusion bodies in the glandular epithelial cells, inflammatory cell infiltrations in the lamina propria, submucosa, and a muscle layer were seen in the gizzards. Immunohistochemical staining showed evidence of FAV antigens in the intranuclear inclusion bodies. These findings were recognized regardless of their maternal antibody levels for FAV serotype-1. Gizzard lesions appeared later in the lower-dose-inoculated chickens than in the higher-dose-inoculated chickens. Numerous CD3-positive cells and IgY-positive plasma cells were seen in the gizzard lesions. In 5-week-old chickens the heterophil infiltrations in the lesions were milder than in younger chickens. Intranuclear inclusion bodies also were observed in the epithelial cells of the ileum or cecal tonsils of some chickens. Thus, this study shows that FAV-99ZH causes adenoviral gizzard erosion in broiler chickens without hepatic or pancreatic lesions and that cell infiltration is more severe than in dietary gizzard erosions.     

Microscopic postmortem changes in adrenal glands of the domestic fowl

Eighty-four male white leghorn chickens were killed by CO2 gas to determine the type, rate, and sequence of microscopic postmortem changes in the adrenal glands of dry and wet intact carcasses. They were held at 29 or 18 C with 50% relative humidity for different times postmortem. The sequence of microscopic postmortem changes was similar in all chickens except at 18 C, when karyorrhexis of cortical and medullary cells was observed. Cellular changes occurred earlier at 29 C than at 18 C and in dry chickens but not in chickens wet with detergent solution before storage, although slight quantitative and qualitative differences between wet and dry chickens were noted. Medullary cells underwent postmortem changes earlier than cortical cells. Nuclei of medullary cells decreased in size, with chromatin clumping leading to pyknosis, followed by cytoplasmic vacuolation, cellular shrinkage, and finally karyolysis and cell dissociation. Cortical cells had nuclear chromatin marginated, nuclei reduced in size initially, and some nuclear fading, followed by pyknosis and karyolysis. Karyorrhexis was not a prominent feature of cortical and medullary cells, although it occasionally occurred before pyknosis. Cytoplasm of cortical cells remained eosinophilic, granular, and vacuolated, but vacuoles became finer later. Pyknotic medullary cells with vacuolated cytoplasm were observed as early as 3 hr postmortem, regardless of the temperature. Diffuse pyknosis of medullary cells was noted at 18 hr in chickens held at 29 C and at 48 hr in chickens held at 18 C. Marked cortical pyknosis was noted only at 36 hr in wet chickens held at 29 C, when bacterial invasion started. Dry chickens held 36 hr at 29 C had diffuse cellular dissociation, karyolysis and cytoplasmic acidophilia, and marked bacterial invasion. Erythrocytes were pyknotic and had cytoplasmolysis. It was concluded that adrenal glands may still be useful for histopathological examination before 18 hr at 29 C and before 48 hr at 18 C.     

Pathogenesis of rotavirus infection in various age groups of chickens and turkeys: clinical signs and virology

Age-related susceptibility patterns of turkeys, broilers, and specific pathogen-free (SPF) White Leghorn chickens to experimentally induced infection with turkey or chicken rotavirus isolates were compared. The following determinants were evaluated: clinical signs, onset and duration of virus production, viral titers, involvement of intestinal villi in the replication of the virus, and the development of antibodies against the virus. Older turkeys and chickens were more susceptible than were their younger counterparts, turkeys were more susceptible than were broiler and White Leghorn chickens (regardless of age), and broiler chickens were slightly more susceptible than were age-matched White Leghorn chickens. Turkeys developed diarrhea, accompanied by high viral titers within 1 day after inoculation with virus. Viral antigen was found in the epithelial cells of the intestinal villi throughout the intestinal tract and some cells of the cecal tonsils. Antibodies could be detected as early as 4 to 5 days after inoculation. These findings were more pronounced in turkeys inoculated at 112 days of age than in birds inoculated at a younger age. Age-related susceptibility patterns were similar in White Leghorn and broiler chickens. Infection was subclinical in birds less than 56 days old, whereas older birds developed soft feces. Egg production in the White Leghorn chickens decreased after being inoculated with virus at 350 days of age.     

Pathology of specific-pathogen-free chickens inoculated with H5N1 avian influenza viruses isolated in Japan in 2004

Specific-pathogen-free chickens inoculated with H5N1 highly pathogenic avian influenza (HPAI) viruses isolated in Japan in 2004 were investigated pathologically. The chickens inoculated intravenously with the viruses died within 26 hr after inoculation. Macroscopically, minimal necrosis of the tip of the comb, and hemorrhages of the palpebral conjunctiva, liver, cerebellum, and muscles were rarely observed. Histologically, dead chickens had minimal focal necrosis of hepatocytes with fibrinous thrombi in sinusoids, mild necrosis of splenic ellipsoids with fibrinous exudation, minimal necrosis of the brain, mild necrosis of epidermal cells of the comb with congestion of the lamina propria, and hemorrhages and edema of the lamina propria of the conjunctiva. Virus antigens were seen in the sinusoidal endothelial cells and hepatocytes in the liver, the capillary endothelial cells of the spleen, the capillary endothelial cells and cardiac myocytes in the heart, the capillary endothelial cells and necrotic nerve cells in the brain, the capillary endothelial cells in the lamina propria of the comb, the renal tubular epithelial cells, and the pancreatic acinar cells. The chickens inoculated by natural infectious routes died within 1-4 days after inoculation. Macroscopically, some chickens had hemorrhages in the conjunctiva, edematous swelling of the face and wattles, hydropericardium, hemorrhages of the proventriculus and bursa of Fabricius, increased secretion of tracheal mucus, and congestion and edema of lungs. Histologic lesions by natural infectious routes were similar to those by intravenous inoculation, except for the pancreatic necrosis. This study suggests H5N1 HPAI viruses isolated in Japan in 2004 cause pathologic conditions similar to natural cases.     

Natural killer cell activity of chicken intraepithelial leukocytes against rotavirus-infected target cells

Intraepithelial leukocytes (IEL) and splenocytes collected from uninfected and rotavirus-infected chickens were evaluated for cytotoxic activity against a natural killer (NK) cell-susceptible lymphoblastoid cell line (LSCC-RP9) and against rotavirus-infected chick kidney cells in 4-h chromium-release assays. Both splenocytes and IELs from uninfected and rotavirus-infected chickens were cytotoxic for LSCC-RP9, and the levels of this NK cell activity were not altered by infection of the host with rotavirus. IELs but not splenocytes from uninfected and rotavirus-infected chickens were cytotoxic for rotavirus-infected but not for uninfected chick kidney cell targets. Because this cytotoxic activity was not induced nor altered by rotavirus infection of the host, and was not major histocompatibility complex-restricted, it was considered to be due to NK cell activity. The cytotoxicity of IELs against rotavirus-infected target cells was dose-dependent; however, there was some suppression of cytotoxic activity at high effector to target cell ratios. There were no differences in the cytotoxic activities of IELs collected from the duodenum versus the jejunum. The in vitro cytotoxic activity of IELs against rotavirus-infected target cells suggested that NK cell activity may be an important immune response to rotavirus infections in vivo. The absence of cytotoxic activity by splenocytes against rotavirus-infected target cells indicated that there may be different subpopulations of NK cells in the spleen and intestinal epithelium of chickens.     

Donor primordial germ cell-derived offspring from recipient germline chimaeric chickens: Absence of long term immune rejection and effects on sex ratios

1. Germline chimaeric chickens were produced by the transfer of primordial germ cells, and the generation of donor-derived offspring was examined for a maximum of 146 weeks. 2. The frequencies of donor-derived offspring from the chimaeras were 47% to 97%, and no apparent changes in frequency were observed with increasing age during the test period. 3. Differentiation of donor primordial germ cells into functional gametes appeared to be restricted to a degree at some developmental stage in the gonads of chimaeric chickens of the opposite sex.     

Relationship between tumour development and detection of Marek's disease virus in the feather follicular epithelium of older chickens

To demonstrate the relationship between tumour development and virus replication, eight specific-pathogen-free pullets of line P2 (Group P; 14 weeks old) and five adult chickens (Group A; 96 weeks old) were inoculated with virulent Marek's disease virus (vMDV). Five chickens of Group P died or were euthanised due to moribund condition following the development of neoplastic lesions between days 53 and 91. On histopathological examination, these lesions were characterised by the proliferation of lymphoid cells of variable size. On analysis by polymerase chain reaction (PCR), the MDV meq gene was detected in Group P from day 21, and it was continuously identified in five chickens until they died or were euthanised. Abnormal signs and histopathological changes were not observed in chickens of Group A. The MDV meq gene was temporarily detected in some chickens of Group A, but it remained almost undetectable throughout the experimental period. In older chickens inoculated with vMDV, the onset of MD lymphoma development tended to be delayed as compared with the young chicks. The relationship between MD lymphoma development and virus replication in older chickens has been suggested. Our data might indicate the underlying existence of an age-related resistance to vMDV challenge.     

Ultrastructure of parathyroid glands of growing chickens with vitamin D deficiency rickets

Ultrastructural studies were conducted on the parathyroid glands (PTG) of 2- to 7-week-old nontreated control growing broiler chickens (group A, 6 chickens); the same aged chickens with experimentally induced vitamin D deficiency rickets (18 morbid chickens each from groups B, C, and D); and in 4- to 9-week-old chickens naturally affected with rickets (group E, 8 chickens). Four types of principal cells were classified in the PTG of clinically normal birds and in PTG of birds with secondary hyperparathyroidism as follows: Type I cell (small cell in the resting phase); type II cell (medium-sized cell with well-developed rough-surfaced endoplasmic reticulum in the synthesizing phase); type III cell (large cell with well-developed Golgi apparatus and cytoplasmic organelles in the packaging and secretory phase); and type IV cell (medium-sized cell with poor cytoplasmic organelles in the involuting phase). Many coated vesicles were observed in type II and type III cells. In young control chickens, principal cells of the PTG were considered in the resting phase; in older control chickens, there were increased numbers of cells in the synthesizing phase. In the vitamin D deficiency groups, principal cells were in the synthesizing and packaging and secretory phases. In birds with naturally occurring rickets, principal cells were in packaging and secretory phases, with a small number of cells in the involuting phase. The ultrastructural changes of the PTG from the initial to final phases in growing chickens with secondary hyperparathyroidism were demonstrated.