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STAR Protocols · 2024 · IF 1.3

Protocol for the cryopreservation of Cryptosporidium isolates using enteroids

Miner Deng,Yanting Wei,Na Li,Lihua Xiao,Yaoyu Feng,Yaqiong Guo

Summary A major bottleneck in the progress of Cryptosporidium research is the lack of accessible cryopreservation of Cryptosporidium oocysts. Here, we present a protocol for the cryopreservation of Cryptosporidium isolates using enteroids. We describe the steps for the establishment of enteroid cultures and cryopreservation of C. parvum -infected HCT-8 cultures. We then detail procedures for the recovery and propagation of frozen parasites using enteroids. For complete details on the use and execution of this protocol, please refer to Deng et al. 1

CAS / MDL: 145-42-6

研究领域: 微生物学 · 寄生虫学 · 细胞生物学

✓ 已验证产品关联 DOI PubMed 高影响因子

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VETERINARY RESEARCH · 2022 ·Vol. 53 ·期 1 ·1-21 · IF 3.829

A multiepitope vaccine encoding four Eimeria epitopes with PLGA nanospheres: a novel vaccine candidate against coccidiosis in laying chickens

Yu ZhengQing,Chen SiYing,Huang JianMei,Ding WenXi,Chen YuFeng,Su JunZhi,Yan RuoFeng,Xu LiXin,Song XiaoKai,Li XiangRui

With a worldwide distribution, Eimeria spp. could result in serious economic losses to the poultry industry. Due to drug resistance and residues, there are no ideal drugs and vaccines against Eimeria spp. in food animals. In the current study, a bioinformatics approach was employed to design a multiepitope antigen, named NSLC protein, encoding antigenic epitopes of E. necatrix NA4, E. tenella SAG1, E. acervulina LDH, and E. maxima CDPK. Thereafter, the protective immunity of NSLC protein along with five adjuvants and two nanospheres in laying chickens was evaluated. Based on the humoral immunity, cellular immunity, oocyst burden, and the coefficient of growth, the optimum adjuvant was evaluated. Furthermore, the optimum immune route and dosage were also investigated according to the oocyst burden and coefficient of growth. Accompanied by promoted secretion of antibodies and enhanced CD4+ and CD8+ T lymphocyte proportions, NSLC proteins entrapped in PLGA nanospheres were more effective in stimulating protective immunity than other adjuvants or nanospheres, indicating that PLGA nanospheres were the optimum adjuvant for NSLC protein. In addition, a significantly inhibited oocyst burden and growth coefficient promotion were also observed in animals vaccinated with NSLC proteins entrapped in PLGA nanospheres, indicating that the optimum adjuvant for NSLC proteins was PLGA nanospheres. The results also suggested that the intramucosal route with PLGA nanospheres containing 300 μg of NSLC protein was the most efficient approach to induce protective immunity against the four Eimeria species. Collectively, PLGA nanospheres loaded with NSLC antigens are potential vaccine candidates against avian coccidiosis.

CAS / MDL: 57-88-5 85261-20-7 22427-39-0 97281-47-5 7758-29-4

研究领域: 兽医学 · 免疫学 · 纳米技术 · 家禽健康

✓ 已验证产品关联 DOI PubMed 高影响因子

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Protocol for the cryopreservation of Cryptosporidium isolates using enteroids

A multiepitope vaccine encoding four Eimeria epitopes with PLGA nanospheres: a novel vaccine candidate against coccidiosis in laying chickens