Genotyping of Echinococcus granulosus and the evaluation of the extracts of Carica papaya and Juglans regia for their scolicidal effect
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Date
2023-03-17
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Sher-e-Kashmir University of Agricultural Sciences & Technology, Jammu (J&K)
Abstract
The prevalence of cystic echinococcosis was determined in 1254 small ruminants (723 sheep and 531 goats) and 305 dogs, including 122 pet dogs visiting small animal clinics, 183 stray dogs in Jammu division for a period from March, 2021 - August, 2022. Among sheep and goats, 209 animals (sheep = 137 and goats = 72) were found infected with hydatidosis with the overall infection rate of 16.67% (sheep = 18.95% and goats = 13.56%). Among different organs examined, the highest infection rate was recorded in liver as 56.94% (sheep = 59.85% and goats = 51.39%), followed by lungs as 31.58% (sheep = 29.93% and goats = 34.72%) and least as mixed infection in liver and lungs as 11.48% (sheep = 10.22% and goats = 13.89%). The overall fertility rate of hydatid cysts from slaughtered sheep and goats was found to be 66.03%. A total of 23.44% of the cysts were found to be sterile and 10.52% cysts were calcified. In terms of the different cyst size of slaughtered sheep, the fertility rate of hydatid cysts was 60.87%, 75.42% and 50.0% for cyst size <4cm, 4-8cm and >8cm, respectively. In terms of the different cyst size of slaughtered goats, the fertility rate of hydatid cysts was 52.38%, 84.21% and 46.15% for cyst size <4cm, 4-8cm and >8cm, respectively. Among dogs, high prevalence was observed in stray dogs (34.43%) as compared to pet/companion dogs (2.45%). Molecular prevalence based on copro pcr was conducted, in which the overall prevalence of Echinococcus was found to be 14.09% [stray (22.95%); pet (0.82%)]. The ethanolic extracts of Juglans regia and Carica papaya showed significant scolicidal activity against E. granulosus, under in-vitro conditions with reference to the known standard drug “Praziquantel”. Against Juglans regia, highest mortality was observed at 30 mg/ml concentration at different exposure time as 10 min. (88.58%), 20 min. (91.24%), 30 min. (93.16%) and 40 min. (96.64%). Against Carica papaya, highest mortality was observed at 30 mg/ml concentration at different exposure time as 10 min. (82.95%), 20 min. (85.83%), 30 min. (90.23%) and 40 min. (92.95%). For the identification of Echinococcus species from cyst of sheep, goats and humans, a PCR protocol employing the Cox1 primers was used. The amplified products of PCR were electrophoretically separated on agar gels (1%) and analysed in a gel documentation system. Clear amplification was noticed at 440 bp for sheep, goat and human. The eluted products were commercially sequenced in an automatic DNA sequencer at Biokart India Pvt. Ltd. Kasturi Nagar, East of NGEF, Bangalore. A total of five sequences were submitted to NCBI GeneBank. The generated Cox1 partial sequences were assigned the accession numbers as: dog (ON427821), goat (ON427822), sheep (ON427823), human1 (OP215294) and human2 (OP215295). All the Cox1 sequences from Dog, goat, sheep and humans were clustering in same clade, irrespective of their geographical location, representing the cycle of transmission in Jammu region as Echinococcus granulosus G1-G3 complex.
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