The process establishment of raising Toxocara canis

Abstract

    Toxocariasis is a prevalent zoonotic disease which is caused by Toxocara canis larvae or Toxocara cati larvae. Diagnosis of human toxocariasis depends on antibody detection against excretions-secretions (E/S) from Toxocara larvae. Since the earliest work of De Savigny, describing the hatching methods and culturing Toxocara larvae, few advances in the method have been made, but those methods are difficult to follow, expensive and have different results. In this work, we propose a simple and effective method for maximizing fertilized Toxocara canis eggs. Adult worms were cultured in medium to collect fertilized eggs, after 5-7 days, eggs were obtained using uterus excision method. The larvae inside the eggs will be triggered by CO₂ produced by the HBSS reaction. The larvae were then filtered through 40μl diameter sieve to collect larvae. The results showed that the proportion of fertilized eggs from culture method (82.5%) is higher than that of those obtained using uterus excision method (40%). This method proved more effective than the original De Savigny method in three ways: (i) Maximizing the number of larvae collected, (ii) improving the larval purity, and (iii) markedly reducing the execution time and labours of the protocol. E/S Toxocara canis proteins were analyzed by SDS-PAGE electrophoresis , showing at least 17 proteins detected. These proteins have molecular weights from 19.5 to 498.5 kDa and the 33.1 kDa protein has the highest proportion.