Spss 19.0 program

SPSS 19.0 program (SPSS INC., Chicago, IL, USA) was used to process the analysis. One-way analyses of variance was used to analyze variance. The pen was the experimental unit. Multiple comparisons among different treatment groups were tested by Duncan multiple range test. The results were shown as means. The level of statistical significance was set at p<0.05. Linear and quadratic significance were evaluated by regression estimation in regression procedure. When the linear or quadratic p<0.05, the regression equation is generated. According to the curve, the most suitable ABSR supplement level was calculated. In the quadratic regression equation (Y = AX²+BX+C), when Y reaches the maximum value, X = −B/2A.

In the LC₅₀ test, M. japonicus were randomly divided into seven groups; each group had three tanks. Among them, each healthy M. japonicus in six groups was intramuscularly injected with 50 μl of viral inoculum, and the concentrations were 3.95 × 10⁹, 3.95 × 10⁸, 3.95 × 10⁷, 3.95 × 10⁶, 3.95 × 10⁵, and 3.95 × 10⁴ copies/μg DNA, respectively. The other group was injected with 50 μl phosphate-buffered saline (PBS, pH 7.4) instead of the viral inoculum. The cumulative survival rate was counted every 4 h, and dead shrimp were picked up to avoid a secondary infection. The methods of viral inoculum preparation and quantification can be found in previous studies (Qiu et al., 2018 (link); Chen X. et al., 2019 (link)). The DIV1 inoculum was tested by nest PCR to ensure that it was not contaminated with WSSV and IHHNV, and the nest PCR primers, specific qPCR primers, and TaqMan probe for DIV1 detection and quantification are shown in Table 1. The LC₅₀ was calculated by probit analysis on SPSS 19.0 program (SPSS Inc., Chicago, IL, United States) using the Bliss method (Bliss, 1939 (link)).

Male Sprague Dawley rats (250–300 g) and male ICR mice (20–22 g) were purchased from the Laboratory Animal Center of Capital Medical University. All evaluations were based on the protocol reviewed and approved by Ethics Committee of Capital Medical University. The committee assured that the animal welfare was maintained in accordance to the requirements of Animal Welfare Act and NIH Guide for Care and Use of Laboratory Animals. Statistical analyses of all biological data were carried out by use of ANOVA, and LSD for multiple group comparison. All analyses were done with SPSS 19.0 program, and p-value < 0.05 was considered statistically significant. THPDTPI was self-prepared by following our procedure [8 ].

The SPSS19.0 program (SPSS Inc., Chicago, IL, United States) was used for statistical analysis. The data are presented as the means ± SD of at least three independent experiments. Student’s t-test was used for two-group comparisons. p < 0.05 was considered to indicate a statistically significant difference.