The invention relates to the technical field of feed additives, in particular to a special micro-ecological preparation for laying hens and a preparation method of the special micro-ecological preparation. The special micro-ecological preparation for the laying hens comprises the following raw materials: bacillus natto powder, aspergillus niger powder, calcium tartrate and chitosan oligosaccharide, wherein the bacillus natto powder and the aspergillus niger powder are prepared by adopting a solid fermentation method, the calcium tartrate is prepared by decalcifying shrimp and crab shells, andthe chitosan oligosaccharide is prepared by taking the decalcified shrimp and crab shells as a raw material to prepare a fermentation culture medium, inoculating bacillus subtilis and paecilomyces cicadae, and carrying out fermentation degradation to obtain the special micro-ecological preparation. Laying hen feed added with the special micro-ecological preparation for the laying hens is used forfeeding the laying hens, so that the concentrated protein content of obtained eggs can be increased, meanwhile, the eggshell thickness is increased, and the quality of the eggs is remarkably improved.