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两种猪衣原体分离株在无菌猪中引起的肠道病变

Intestinal lesions caused by two swine chlamydial isolates in gnotobiotic pigs.

作者信息

Rogers D G, Andersen A A

机构信息

Department of Veterinary and Biomedical Sciences, University of Nebraska, Lincoln 68583-0907, USA.

出版信息

J Vet Diagn Invest. 1996 Oct;8(4):433-40. doi: 10.1177/104063879600800405.

Abstract

The objective of this study was to determine whether 2 distinct chlamydial isolates recovered from the intestines and feces of diarrheic nursery pigs could cause intestinal lesions in gnotobiotic pigs. Both isolates share biological characteristics with Chlamydia trachomatis. Chlamydial isolates R27 and R19 were propagated in Vero cells or embryonated eggs, respectively, and suspended in sucrose-phosphate-glutamine buffer with 10% fetal bovine serum for inoculation. Sham inocula were prepared from uninfected cell culture lysates and from uninfected eggs. Each piglet was fed 1 ml of inoculum or sham inoculum at 3-4 days of age. Ten piglets were each fed 10(9) inclusion-forming-units (IFU) and 14 piglets were each fed 10(6) IFU of isolate R27; 5 control piglets were fed sham inoculum. Twenty piglets were each fed 10(5) IFU R19; 5 control piglets were fed sham inoculum. All infected piglets developed diarrhea 4-5 days postinfection (DPI). Most piglets fed 10(9) IFU R27 became anorexic, dehydrated, and weak and were necropsied 4-7 DPI. Piglets fed 10(6) IFU R27 or 10(5) IFU R19 were necropsied 4, 7, 10, 14, and 18 DPI. Diarrhea, although never profuse, persisted in the piglets fed 10(6) IFU R27 or 10(5) IFU R19 through 12 DPI. At necropsy, all diarrheic piglets had watery colonic contents with flecks of undigested curd. In small intestine, histologic lesions were seen most consistently in distal jejunum and ileum. Distal jejunum and ileum from piglets fed 10(9) IFU R27 and necropsied 4-5 DPI were characterized by villus atrophy and multifocal necrosis of villi; necrosis was limited to the tips or apical one half of villi. Mild to severe villus atrophy, lymphangitis, and perilymphangitis were seen in the distal jejunum and ileum from all infected piglets 7 and 10 DPI. Colon from 1 infected piglet necropsied 10 DPI had mild focal serositis; significant colonic lesions were not seen in the other infected piglets. Immunostaining done on sections of distal jejunum and ileum revealed chlamydial antigen in villus enterocytes, occasional goblet cells, and occasional cryptal enterocytes and in foci of lymphangitis and perilymphangitis; the amount of detectable chlamydial antigen decreased after 4 DPI. In colon, sparse positive staining was seen in surface enterocytes and cryptal enterocytes. Ultrastructural examination of ileal villus enterocytes revealed chlamydiae, often together with glycogen particles, in vacuoles or occasionally free in the cytoplasm. The results indicated that the swine chlamydial isolates used in this study are intestinal pathogens in gnotobiotic pigs.

摘要

本研究的目的是确定从腹泻仔猪的肠道和粪便中分离出的两种不同衣原体菌株是否会在无菌仔猪中引起肠道病变。这两种菌株均具有沙眼衣原体的生物学特性。衣原体菌株R27和R19分别在Vero细胞或鸡胚中繁殖,并悬浮于含10%胎牛血清的蔗糖-磷酸盐-谷氨酰胺缓冲液中用于接种。假接种物由未感染的细胞培养裂解物和未感染的鸡胚制备。每头仔猪在3 - 4日龄时喂食1 ml接种物或假接种物。10头仔猪每头喂食10⁹包涵体形成单位(IFU),14头仔猪每头喂食10⁶ IFU的菌株R27;5头对照仔猪喂食假接种物。20头仔猪每头喂食10⁵ IFU R19;5头对照仔猪喂食假接种物。所有感染仔猪在感染后4 - 5天(DPI)出现腹泻。大多数喂食10⁹ IFU R27的仔猪出现厌食、脱水和虚弱,并在4 - 7 DPI时进行剖检。喂食10⁶ IFU R27或10⁵ IFU R19的仔猪在4、7、10、14和18 DPI时进行剖检。喂食10⁶ IFU R27或10⁵ IFU R19的仔猪腹泻虽不严重,但持续到12 DPI。剖检时,所有腹泻仔猪的结肠内容物呈水样,有未消化凝乳块。在小肠中,组织学病变最常见于空肠远端和回肠。喂食10⁹ IFU R27并在4 - 5 DPI剖检的仔猪空肠远端和回肠表现为绒毛萎缩和绒毛多灶性坏死;坏死仅限于绒毛尖端或顶端的一半。所有感染仔猪在7和10 DPI时,空肠远端和回肠出现轻度至重度绒毛萎缩、淋巴管炎和淋巴管周围炎。1头在10 DPI剖检的感染仔猪的结肠有轻度局灶性浆膜炎;其他感染仔猪未见明显结肠病变。对空肠远端和回肠切片进行免疫染色显示,绒毛肠上皮细胞、偶尔的杯状细胞、偶尔的隐窝肠上皮细胞以及淋巴管炎和淋巴管周围炎病灶中有衣原体抗原;4 DPI后可检测到的衣原体抗原量减少。在结肠中,表面肠上皮细胞和隐窝肠上皮细胞可见稀疏的阳性染色。对回肠绒毛肠上皮细胞的超微结构检查显示,衣原体常与糖原颗粒一起存在于液泡中,偶尔游离于细胞质中。结果表明,本研究中使用的猪衣原体菌株是无菌仔猪的肠道病原体。

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