A simple, robust and semi-automated parasite egg isolation protocol

被引:53
作者
Mes, Ted H. M. [1 ]
Eysker, Maarten [2 ]
Ploeger, Harm W. [2 ]
机构
[1] NIOO KNAW, Netherlands Inst Ecol, Ctr Estuarine & Marine Ecol, NL-4400 AC Yerseke, Netherlands
[2] Univ Utrecht, Fac Vet Med, Dept Immunol & Infect Dis, NL-3508 TD Utrecht, Netherlands
关键词
DAIRY-CATTLE; DENSITY;
D O I
10.1038/nprot.2007.56
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Large-scale parasite quantification is required for improving our understanding of the epidemiology and genetics of host-parasite interactions. We describe a protocol that uses a low-density salt solution for flotation and centrifugation of nematode eggs. Subsequently, sucrose flotation and precipitation are used to obtain clear egg preparations. Most traditional quantification protocols such as the McMaster technique are unsuited for the standardized processing of large numbers of samples and the analysis of large amounts of feces per sample. Consequently, they are suited only for small-scale surveys. Our protocol, which can be used to analyze up to 6 g of feces, results in clear egg preparations that are concentrated in wells of a microtiter plate and that are suited for digital recording and automated counting. Starting from a fecal suspension in the first flotation solution to a digital recording requires approximately 40 min per 24 samples.
引用
收藏
页码:486 / 489
页数:4
相关论文
共 7 条
[1]   Nematode parasitism in adult dairy cows in Belgium [J].
Agneessens, J ;
Claerebout, E ;
Dorny, P ;
Borgsteede, FHM ;
Vercruysse, J .
VETERINARY PARASITOLOGY, 2000, 90 (1-2) :83-92
[2]   Nematode parasites of adult dairy cattle in the Netherlands [J].
Borgsteede, FHM ;
Tibben, J ;
Cornelissen, JBWJ ;
Agneessens, J ;
Gaasenbeek, CPH .
VETERINARY PARASITOLOGY, 2000, 89 (04) :287-296
[3]  
COX DD, 1962, J AM VET MED ASSOC, V141, P706
[4]   DETERMINATION OF THE SPECIFIC-GRAVITY OF CERTAIN HELMINTH EGGS USING SUCROSE DENSITY GRADIENT CENTRIFUGATION [J].
DAVID, ED ;
LINDQUIST, WD .
JOURNAL OF PARASITOLOGY, 1982, 68 (05) :916-919
[5]  
*MAFF, 1977, MAN VET PAR LAB TECH, V18
[6]   A novel method for the isolation of gastro-intestinal nematode eggs that allows automated analysis of digital images of egg preparations and high throughput screening [J].
Mes, THM ;
Ploeger, HW ;
Terlou, M ;
Kooyman, FNJ ;
Van der Ploeg, MPJ ;
Eysker, M .
PARASITOLOGY, 2001, 123 :309-314
[7]  
NIBLACK TL, 1993, J NEMATOL, V25, P880