Abstract
Several methods have been proposed for separation of eimerian oocysts and trichostrongylid eggs from extraneous debris; however, these methods have been considered to be still inconvenient in terms of time and wide-ranging applications. We describe herein an alternative way using the combination of electrical cream separator and vacuum filtration for harvesting and purifying eimerian oocysts and haemonchine eggs on large-scale applications with approximately 81% and 92% recovery rates for oocysts and nematode eggs obtained from avian and ovine faeces, correspondingly. The sporulation percentages as a measure of viability in the harvested oocysts and eggs from dry faecal materials are nearly 68% and 74%, respectively, and 12 liters of faecal suspension can be processed in approximately 7.5 min. The mode of separation in terms of costs (i.e. simple laboratory equipments and comparably cheap reagents) and benefits renders the reported procedure an appropriate pursuit to harvest and purify parasite oocysts and eggs on a large scale in the shortest duration from diverse volumes of environmental samples compared to the modified traditional sucrose gradient, which can be employed on a small scale.
Highlights
A recent study by[13] has employed two-step method, sucrose flotation followed by density gradient centrifugation exploiting the lymphocyte separation medium (LSM) to produce highly-purified nematode eggs
Animal experiments were conducted in accordance with the guidelines of Beijing the Municipality on the Review of Welfare and Ethics of Laboratory Animals approved by the Beijing Municipality Administration Office of Laboratory Animals (BAOLA), and under the protocol (CAU-AEC-2010–0603) approved by the China Agricultural University Animal Ethics Committee
Two-week-old specific pathogen free (SPF) Arbor Acres (AA) broiler chicks were purchased from Beijing Arbor Acres Poultry Breeding Co., Ltd
Summary
Large-scale method for separating oocysts involving sedimentation of heavier particles from faecal suspension, collection of oocysts from the supernatant, repeated washing and settling in potassium dichromate (K2Cr2O7) solution. Eckert et al.[16] have lengthily described typical laboratory methods involving screening, sedimentation and gravitational salt flotation for oocyst isolation from faeces This technique is for the concentration and purification of oocysts from large quantities of faecal samples, necessitates long periods of time and some concerns about the efficiency of oocyst and egg recovery rates (unpublished data). The overall concern in all conventional methods has been to develop an efficient procedure for viable oocyst and egg isolation, which can be applied on a large scale in the shortest duration Driven by this need, we proposed here a simple two-step procedure for obtaining highly purified oocysts and nematode eggs. A modified discontinuous (step-gradients) sucrose gradient method was employed for the isolation of eimerian oocysts and haemonchine eggs
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
