Observation on antinematode activity of the recombinant Bacillus thuringiensis-derived Cry5B against the giant panda Baylisascaris schroederi in vitro

HUANG Wenjun; LUO Li; CHEN Xin; LIU Li; LIAO Lihui; WANG Xiaolan; LI Bi; LI Mingxi; CHEN Min; YI Dejiao; LI Han; ZHANG Hao; ZHUO Guifu; LIU Yunjian; LI Yingxin; CHEN Yijun; ZHOU Xuan; XIE Yue

doi:10.16829/j.slxb.150643

2022 , Vol. 42 >Issue 3: 304 - 311

DOI: https://doi.org/10.16829/j.slxb.150643

ORIGINAL PAPERS

Observation on antinematode activity of the recombinant Bacillus thuringiensis-derived Cry5B against the giant panda Baylisascaris schroederi in vitro

HUANG Wenjun ,
LUO Li ,
CHEN Xin ,
LIU Li ,
LIAO Lihui ,
WANG Xiaolan ,
LI Bi ,
LI Mingxi ,
CHEN Min ,
YI Dejiao ,
LI Han ,
ZHANG Hao ,
ZHUO Guifu ,
LIU Yunjian ,
LI Yingxin ,
CHEN Yijun ,
ZHOU Xuan ,
XIE Yue

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1. Veterinary Hospital, Chengdu Research Base of Giant Panda Breeding, Chengdu 610081, China;
2. College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang 611130, China

Received date: 2021-09-13

Revised date: 2022-02-23

Online published: 2022-06-02

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Abstract

The roundworm Baylisascaris schroederi is one of the most serious intestinal parasitic nematodes found in the giant panda, a rare species endemic to China. Given the current situation of the long-term anthelmintic treatment-causing resistance and ecological pollution, Bacillus thuringiensis (Bt)-producing crystal protein Cry5B is an ideal and novel antiascariasis drug candidate due to its specific antinematode activity. This study was designed to prokaryotically produce the recombinant protein Cry5B from the Bt YBT-1518 and to evaluate its antinematode activity on the intestinal fourthstage larvae (L4s) and adults of B. schroederi. The results showed that the recombinant Bt YBT-1518 Cry5B protein consisted of 1 246 amino acids with a molecular weight (MW) of 139. 889 kDa, contained Endotoxin_N, δ-Endotoxin_C, Endotoxin_C, Endotoxin_C2, and Cry1Ac_D5 domains, and shared the closest relationship with that of Bt PS86Q3. After optimization, the recombinant Cry5B protein was expressed in the supernatant with the maximum yield when IPTG was up to 1. 4 mmol/L. Further analysis of antinematode activity indicated a significant dose-dependent response to the recombinant Cry5B protein with ED₅₀ of 14. 5 μg/mL on day 3 and 0. 16 μg/mL on day 7 for L4s. It appeared that adults were more sensitive to this protein than L4s as all worms became immotile on day 2 and even dead on day 7. Combined, these findings suggested that the recombinant Bt YBT-1518 Cry5B protein is able to completely intoxicate B. schroederi. These results provided insights into the development and clinical application of Bt YBT-1518 recombinant Cry5B protein as a new drug against the giant panda baylisascariasis.

Key words： Giant panda; Bacillus thuringiensis; Baylisascaris schroederi; Cry5B; Antinematode activity

Cite this article

HUANG Wenjun , LUO Li , CHEN Xin , LIU Li , LIAO Lihui , WANG Xiaolan , LI Bi , LI Mingxi , CHEN Min , YI Dejiao , LI Han , ZHANG Hao , ZHUO Guifu , LIU Yunjian , LI Yingxin , CHEN Yijun , ZHOU Xuan , XIE Yue . Observation on antinematode activity of the recombinant Bacillus thuringiensis-derived Cry5B against the giant panda Baylisascaris schroederi in vitro[J]. ACTA THERIOLOGICA SINICA, 2022 , 42(3) : 304 -311 . DOI: 10.16829/j.slxb.150643

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