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Frontiers of Chemical Science and Engineering

ISSN 2095-0179

ISSN 2095-0187(Online)

CN 11-5981/TQ

Postal Subscription Code 80-969

2018 Impact Factor: 2.809

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Front. Chem. Sci. Eng.    2019, Vol. 13 Issue (2) : 377-384    https://doi.org/10.1007/s11705-018-1777-9
RESEARCH ARTICLE
Safety evaluation of microbial pesticide (HaNPV) based on PCR method
Miao Zhao1, Shufei Li1, Qinghong Zhou1, Dianming Zhou1,2, Ning He1, Zhiyong Qian1()
1. Departmnet of Toxicology, Tianjin Centers for Disease Control and Prevention, Tianjin 300011, China
2. State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082, China
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Abstract

Microbial pesticides can prevent and control diseases and pests of crops, and has become one of the important measures to ensure food and environmental safety. However, the potential harm of microbial pesticides to humans and animals is a serious concern at home and abroad. In this paper, we have investigated the infectivity and pathogenicity of a representative of viral microbial pesticides, helicoverpa armigera nuclear polyhedrosis virus (HaNPV), by specific and highly sensitive polymerase chain reaction technology. The results show that HaNPV can be gradually cleared in a short time after getting into blood of experimental rats, and does not infect other tissues or organs of animals; also indicate that the test subjects are not infectious to experimental rats after intravenous injection of HaNPV. Our method has good specificity and repeatability, and could provide an important reference for establishment of infectivity and pathogenicity detection methods for viral microbial pesticides in future.
Keywords microbial pesticides      HaNPV      acute intravenous injection      infectivity      pathogenicity     
Corresponding Author(s): Zhiyong Qian   
Online First Date: 25 February 2019    Issue Date: 22 May 2019
 Cite this article:   
Miao Zhao,Shufei Li,Qinghong Zhou, et al. Safety evaluation of microbial pesticide (HaNPV) based on PCR method[J]. Front. Chem. Sci. Eng., 2019, 13(2): 377-384.
 URL:  
https://academic.hep.com.cn/fcse/EN/10.1007/s11705-018-1777-9
https://academic.hep.com.cn/fcse/EN/Y2019/V13/I2/377
Groups Sex Day 0 Day 7 Day 14 Day 21
Experimental Males 213.0±3.4 262.2±3.3 312.8±8.3 329.6±6.7
Females 195.6±8.8 227.4±5.5 252.4±9.3 285.2±4.5
Control Males 210.6±7.0 263.2±7.8 301.0±7.3 323.8±5.9
Females 194.2±9.8 222.4±6.3 256.6±5.6 278.8±9.0
Tab.1  Effects of intravenous injection with HaNPV on body weight (n = 5, x±s, g)
Fig.1  PCR amplification products electrophoresis of HaNPV in tissues, blood and cecum contents of rats dissected immediately after exposure. M: marker; 1: positive control; 2?9 (male): brain, blood, lymph nodes, liver, spleen, lung, kidney, cecum content; 10?17 (female): brain, blood, lymph node, liver, spleen lung, kidney, cecum content
Fig.2  PCR amplification products electrophoresis of HaNPV in tissues, blood and cecum contents of rats dissected after exposure for 3 days. Loading order of lanes is the same as Fig. 1
Fig.3  PCR amplification products electrophoresis of HaNPV in tissues, blood and cecum contents of rats dissected after exposure for 7 days. Loading order of lanes is the same as Fig. 1
Fig.4  PCR amplification products electrophoresis of HaNPV in tissues, blood and cecum contents of rats dissected after exposure for 14 days. Loading order of lanes is the same as Fig. 1
Fig.5  PCR amplification products electrophoresis of HaNPV in tissues, blood and cecum contents of rats dissected after exposure for 21 days. Loading order of lanes is the same as Fig. 1
Fig.6  PCR amplification products electrophoresis of HaNPV in tissues, blood and cecum contents of rats in the control group which was dissected on the 21st day after experiment began. Loading order of lanes is the same as Fig. 1
Groups Sex White blood cell count /×109 L–1 Red blood cell count
/×1012 L–1		 Hemoglobin
/(g?L–1)		 Neutrophils /% Lymphocyte /% Monocytes /% Eosinophil /% Basophils /%
Experimental Males 9.25±0.46 7.89±0.21 158.40±3.29 17.54±2.18 73.76±2.60 6.96±0.85 1.56±0.32 0.02±0.04
Females 9.24±1.04 7.71±0.46 157.80±3.56 16.36±1.98 74.62±4.13 6.82±0.58 1.46±0.27 0.02±0.04
Control Males 9.04±0.47 7.69±0.37 155.40±5.22 16.14±4.52 75.68±6.04 6.76±1.35 1.16±0.24 0.06±0.05
Females 8.92±0.41 7.79±0.47 156.80±6.72 15.66±4.26 76.40±4.67 6.60±1.68 1.22±0.30 0.04±0.05
Tab.2  Effects of tested substances on hematological parameters (n = 5, x+ s)
Groups Sex ALT
/(U?L–1)		 AST
/(U?L–1)		 TP
/(g?L–1)		 ALB
/(g?L–1)		 GLU
/(mmol?L–1)		 BUN
/(mmol?L–1)		 CRE
/(µmol?L–1)		 CHO
/(mmol?L–1)		 TG
/(mmol?L–1)
Experimental Males 40.8±5.8 135.4±23.5 70.6±2.2 48.2±2.6 4.59±0.26 5.83±0.94 34.8±2.6 2.01±0.25 0.81±0.21
Females 41.2±7.4 134.2±13.3 70.2±5.1 48.8±2.5 4.61±0.30 5.70±1.22 35.6±3.3 2.15±0.39 0.77±0.15
Control Males 32.2±3.5 127.8 × 10.7 67.6±2.6 47.7±1.8 5.13±0.60 5.57±1.89 34.6±2.3 2.09±0.49 0.75±0.19
Females 31.0±3.2 129.4±8.0 68.9±2.9 47.5±3.0 5.20±0.58 5.56±1.47 34.2±3.7 2.13±0.41 0.70±0.20
Tab.3  Effects of tested substances on biochemical parameters (n = 5, x+ s)
Fig.7  Histopathological morphology of rats infected with HaNPV and untreated (HE stained, 400 ×). A?H (the control group): brain, heart, liver, spleen, lungs, kidney, stomach, lymph nodes; I?P (the experimental group): brain, heart, liver, spleen, lungs, kidney, stomach, lymph nodes
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