IEg67 kDa Bovine Hydatid Cyst Antigen: A Candidate for Developing Sero-Diagnostic Assays for Cystic Echinococcosis, a Disease of One Health Importance

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Abstract
Cystic echinococcosis (hydatidosis) is a world-wide zoonotic disease of mainly humans, livestock and dogs, caused by Echinococcus granulosus. The disease can negatively impact food production and animal welfare and causes socio-economic hardship. Here, we aimed to identify the local bovine hydatid cyst fluid (BHCF) antigen for developing a sero-diagnostic assay to be used for the pre-slaughter screening of food animals. In total, 264 bovines approved for slaughter in Pakistan were subjected to serum collection and post-mortem screening for hydatid cysts. These cysts were assessed microscopically to assess fertility and viability, and by PCR for molecular confirmation of species. A BHCF antigen was identified from positive sera via SDS-PAGE, confirmed by Western blot, and quantified via a bicinchoninic acid (BCA) assay. The quantified crude BHCF antigen (iEg67 kDa) was then used in ELISA screening to test all sera collected from known positive and negative animals based on hydatid cyst presence/absence. Of the 264 bovines examined, 38 (14.4%) showed hydatid cysts during post-mortem examination. All of these individuals, plus an additional 14 (total: 52; 19.6%) tested positive based on less time-consuming ELISA examination. Based on ELISA, occurrence in females (18.8%) was significantly higher than in males (9.2%) and was higher in cattle (19.5%) compared to buffalo (9.5%). The infection rate increased with age in both host species: cumulatively, 3.6% in animals aged 2–3 years, 14.6% in 4–5-year-olds and 25.6% in 6–7-year-olds. The occurrence of cysts in cattle was significantly higher in the lungs (14.1%) compared to their livers (5.5%), whereas the opposite was true in buffalo (6.6% livers, 2.9% lungs). For both host species, most cysts in the lungs were fertile (65%), while the majority in the liver were sterile (71.4%). We conclude that the identified iEg67 kDa antigen is a strong candidate for the development of a sero-diagnostic screening assay for the pre-slaughter diagnosis of hydatidosis.
Funding Information
  • International Research Support Initiative Program (1-8/HEC/HRD/2022/12612/IRSIP, NRPU-7018, GCF-273, PARB-18-476, PHEC/ARA/PIRCA/2020-6/8)
  • HEC-National Research Program for Universities (1-8/HEC/HRD/2022/12612/IRSIP, NRPU-7018, GCF-273, PARB-18-476, PHEC/ARA/PIRCA/2020-6/8)
  • HEC-Grand Challenge Fund (1-8/HEC/HRD/2022/12612/IRSIP, NRPU-7018, GCF-273, PARB-18-476, PHEC/ARA/PIRCA/2020-6/8)
  • Pakistan Agriculture Research Board (1-8/HEC/HRD/2022/12612/IRSIP, NRPU-7018, GCF-273, PARB-18-476, PHEC/ARA/PIRCA/2020-6/8)
  • Punjab Higher Education Commission (1-8/HEC/HRD/2022/12612/IRSIP, NRPU-7018, GCF-273, PARB-18-476, PHEC/ARA/PIRCA/2020-6/8)

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