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马体内包囊圆线虫幼虫定量的两种技术比较。

Comparison of two techniques for quantitation of encysted cyathostome larvae in the horse.

作者信息

Reinemeyer C R, Herd R P

出版信息

Am J Vet Res. 1986 Mar;47(3):507-9.

PMID:3963552
Abstract

Haustral portions of intestine of 6 horses were isolated by excising the taeniae coli from the cecum and the ventral colon. Uniform 5-cm X 5-cm sections were cut from the haustra and were illuminated from the serosal side with a strong light source (mural transillumination). Cyathostome larvae encysted in the mucosa and submucosa were observed at 15 X magnification and counted. Two separate counts of the larvae in 80 replicates of tissue by the mural transillumination technique (MTT) revealed no significant (P less than 0.05) difference between sample means. Larvae in tissue sections were counted in situ by MTT, and the mucosal scrapings of the tissue sections were digested in pepsin and HCl to determine larval yields for comparison with the MTT counts. Numbers of larvae recovered by pepsin and HCl digestion for 3 and 6 hours were significantly (P less than 0.01) lower than were numbers originally determined by MTT. Larvae recovered by tissue digestion for 3 or 6 hours were examined individually and given objective scores for morphologic damage. Distribution of scores was time-dependent; increased damage to larvae was associated with a longer time of digestion. Individual 4th-stage cyathostome larvae were dissected from cysts in the large intestinal wall and were incubated in water, 0.9% saline solution, 1.1% HCl, or pepsin (7,000 U of activity/ml). Significantly fewer (P less than 0.05) larvae were recovered from all solutions after 3 and 6 hours. The proportion of dissected larvae that were given high scores after exposure to pepsin was significantly (P less than 0.01) greater than were those held in HCl, saline solution, or water for both periods.

摘要

通过从盲肠和腹侧结肠切除结肠带,分离出6匹马的肠袋状部。从肠袋状部切取大小均一的5厘米×5厘米切片,从浆膜侧用强光源照射(肠壁透照)。在15倍放大倍数下观察并计数包囊在黏膜和黏膜下层的杯状线虫幼虫。通过肠壁透照技术(MTT)对80个组织样本进行两次单独的幼虫计数,结果显示样本均值之间无显著差异(P<0.05)。通过MTT对组织切片中的幼虫进行原位计数,并将组织切片的黏膜刮屑用胃蛋白酶和盐酸消化,以确定幼虫产量,与MTT计数结果进行比较。经胃蛋白酶和盐酸消化3小时和6小时后回收的幼虫数量显著低于(P<0.01)最初由MTT确定的数量。对经3小时或6小时组织消化回收的幼虫进行个体检查,并给予形态损伤客观评分。评分分布与时间相关;幼虫损伤增加与消化时间延长有关。从大肠壁囊肿中解剖出单个第四期杯状线虫幼虫,分别置于水、0.9%盐溶液、1.1%盐酸或胃蛋白酶(活性7000 U/ml)中孵育。3小时和6小时后,从所有溶液中回收的幼虫数量均显著减少(P<0.05)。暴露于胃蛋白酶后获得高分的解剖幼虫比例在两个时间段均显著高于(P<0.01)置于盐酸、盐溶液或水中的幼虫。

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