• Vaccine Name: E. tenella DNA vaccine EtMIC2
• Target Pathogen: Eimeria tenella
• Target Disease: Hemorrhagic cecal coccidiosis
• Vaccine Ontology ID: VO_0011471
• Type: DNA vaccine
• Status: Research
• Antigen: E. tenella microneme 2 (MIC2)
• MIC2 gene engineering:
  • Type: DNA vaccine construction
  • Description: The EtMIC2 coding sequence was subcloned from EtMIC2-pGEX into the BamHI/HindIII sites of the pMal4c vector with a NH2-terminal maltose-binding protein tag, expressed in E. coli in TY broth (20 g/l tryptone, 10 g/l yeast extract, 10 g/l NaCl) containing 100 μg/ml ampicillin, the bacteria grown to OD600 = 0.5, induced with 1.0 mM isopropyl-β-d-thiogalactopyranoside for 3 h at 37 °C, collected by centrifugation, and disrupted by sonication on ice (Misonix, Farmingdale, NY). The EtMIC2 protein was isolated on an amylose affinity column (New England Biolabs, Beverly, MA) according to the manufacturer's instructions, digested with Factor Xa to release EtMIC2, and re-passed through the amylose column to remove maltose-binding protein (Ding et al., 2005).
  • Detailed Gene Information: Click Here.
• DNA vaccine plasmid:
  • DNA vaccine plasmid name:
  • DNA vaccine plasmid VO ID: VO_0005048
• Immunization Route: in ovo

Chicken Response

• Vaccination Protocol: To assess anti-EtMIC2 antibody titers and protective immunity to coccidiosis following in ovo vaccination with the EtMIC2 gene, 75 fertile eggs were distributed into five groups (15/group) and non-injected or injected with 100 μl of sterile PBS, 50 μg/egg of the pcDNA empty vector, or 25 or 50 μg/egg of EtMIC2-pcDNA. To determine the effects of post-vaccination boosting with the EtMIC2 gene, 150 fertile eggs were distributed into 10 groups (15/group) and non-injected or injected with 100 μl of sterile PBS, 50 μg/egg of the pcDNA empty vector, or 25 or 50 μg/egg of EtMIC2-pcDNA (Ding et al., 2005).
• Challenge Protocol: At day 11 post-hatching, chickens were infected with 10,000 sporulated oocysts of E. tenella. Serum samples were collected at days 1, 10, and 17 post-infection (days 10, 21, and 28 post-hatching) and anti-EtMIC2 antibody titers determined by ELISA (Ding et al., 2005).
• Efficacy: Birds vaccinated with Eimeria tenella microneme recombinant gene (EtMIC2) and encoded protein developed protective immunity against infection by E. tenella as assessed by significantly increased body weight gain and decreased fecal oocyst shedding compared with non-vaccinated controls. Vaccination with the EtMIC2 gene also led to protective immunity against infection by E. acervulina, but not E. maxima (Ding et al., 2005).