Study On Chicken Coccidian Oocyst Separation,Collection And Culture In Vitro

To investigate the fast, high recovery, low cost, high purity, sterile collecting methods for coccidia oocysts from chicken manure and to provide foundation and device for producing coccidia vaccine, chicken were fed with different sizes of food and chicken’s manure was dealed with immersing in washing liquid, floating,centrifuging and filtering in different materials and conditions to get oocysts. Collected oocysts were cultured in different nutrient liquid to determin the mature rates and the presence of bacteria. Results of experiment indicated that:(1) Oocyst productions of groups with complete feed,1:1mixed by cracked corn and complete feed and cracked corn decreased gradually. In contrast, recoveries were increased gradually.(2) The recoveries of oocyst didn’t have significant gap (P>0.05) between using cooled boiled water which was10times more than the amount of manure and contained0.5%detergent and2.5%acetic acid and PBS which was10times more than the amount of manure and contained0.5%detergent and2.5%acetic acid. The recoveries of both groups were highly significantly (P<0.01) higher than other soaked and diluted liquid. We found that the recovery of oocyst reached to99.5%, when centrifugal power was F=693g for9min.(3) Using saturated brine to float precipitate containing oocyst was better than other floating medium, which suggested that the recovery of oocyst was88%with centrifugal power.F=209g for3min.The recovery rates of oocysts filtered with5μm and1Oμm pore size membrane had no significant difference (P>0.05), but the recovery rates of both groups were highly significant (P<0.01) higher than the15μm pore size membrane, collected oocysts more pure and up to79.6%.(4) Oocysts maturation rate was up to over95%after72h when they were cultured in aqueous solution containing10%citric acid,0.25%propionic acid,0.75%peroxide and2.5%dichromate solution, while no bacteria could be detected. Oocysts maturation rate got over80%after72h when they were cultured in1.2%to2.4%BCDMH, and bacteria could be detected.The results showed that chicken fed with complete feed had higher oocyst production than with cracked corn and chicken fed with powder had lower oocyst recovery than with larger size food. Manure was soaked and diluted by cooled boiled water which was10times more than the amount of manure and contained0.5%detergent and2.5%acetic acid, filtered after30min by18,80,200mesh filter in turn, then centrifuged by F=693g for9min. Precipitate containing oocyst was floated by saturated brine, centrifuged by F=209g for3min. Oocysts were collected with10μm pore size membrane and cultured in aqueous solution containing10%citric acid,0.25%propionic acid,0.75%peroxide and2.5%dichromate solution. High purity, sterile oocysts were fast, high recovery, low cost obtained from fecal.