[Objective] This study was conducted to explore the method of establishing Lipopolysaccharide (LPS)-induced mastitis pathological models and reveal dynamic pathological changes of mammary tissue before and after LPS perfusion,finally providing convenient animal models for establishing LPS-induced acute clinical mastitis researches.[Method] Twelve lactating rabbits (the 7th day after parturition) were perfused with LPS into the fourth mammary gland via the teat duct.Exactly at 2h before perfusion,6 h,12 h,24 h,48 h,72 h,5 d and 7 d after,the indexes such as rectal temperature,total white blood cells and neutrophils,C-response protein (CPR) content and lactate dehydrogenase (LDH) activity were determined,respectively.Then the rabbits were euthanatized and the mammary glands were removed and fixed for histopathologic evaluations.[Result] The results showed that inflammatory cells were observed in mammary tissue 6 h after LPS perfusion,structure of mammary tissue disorderly at 24 h,self-regeneration after 48 h and back to normal at 7 d.LDH activity was increasing significantly (P<0.05) at 12 h (P<0.05),peaking at 24 h,decreasing at 5 d (P<0.05) and returning to health at 7 d.CRP content was increasing significantly (P<0.05) at 6 to 72 h,peaking at 12 h,decreasing at 48 h,back to normal at 5 d.[Conclusion] The results suggested that the rabbit experimental mastitis model was successfully constructed by LPS perfusion via teat duct.
CAO Sui-zhongYANG De-yingYAO Xue-pingYU Shu-minSHEN Liu-hongLIU Chang-song
[Objective] This study aimed to prepare dairy cow anti-S100A12 antisera and develop a highly effective and sensitive immunological detection reagent for further investigation of the functions of dairy cow S100A12.[Method] Purified S100A12 protein was respectively emulsified with Freund's complete adjuvant and Freund's incomplete adjuvant as the antigen for immunizing New Zealand white rabbits to prepare the polyclonal antisera.The titer was detected using agar double diffusion assay and indirect enzyme-linked immunosorbent assay (ELISA) and the specificity was determined with Western Blot.[Result] The titer of anti-S100A12 antisera was 1∶8 as determined by agar double diffusion assay and over 1∶409 600 by ELISA.Western Blot result showed that the polyclonal antisera could be specifically combined with S100A12 protein.[Conclusion] The results indicated that anti-S100A12 polyclonal antibody with high titer and high specificity was successfully obtained, which provided a novel tool for further investigation of the functions of S100A12 gene.
