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African Journal of Pig Farming ISSN 2375-0731 Vol. 3 (3), pp. 001-005, March, 2015. Available online at 
www.internationalscholarsjournals.org © International Scholars Journals 

 

Author(s) retain the copyright of this article. 

 

Full Length Research Paper 

 

Effects of immune synergist of Chinese medicinal 
herbs on the efficacy of vaccination against classic 

swine fever 

 
Fuchuan Wang1, Yibo Yan1*, Yuhuan Zhang1, Yichao Han1 and Chao Guan2

 
 

1
Institute of Animal Husbandry and Veterinary Sciences, Shanxi Academy of Agricultural Sciences, Taiyuan 030032, China. 

2
Modern Agricultural Research Center, Taiyuan 030000, China. 

 
Accepted 16 November, 2014 

 
Two-month-old piglets were fed with 1, 1.5 and 2% immune synergist of Chinese medicinal herbs 
together with vaccination against classic swine fever. Serum IgG and IgM levels increased more than 
the control group on day 30 (P<0.05). B and T lymphocyte proliferation in piglets fed with 1.5 and 2% 
herbal immune synergist markedly increased on day 30 as compared to the control group (P<0.05). 
Improvement was also observed in T lymphocyte CD3+, CD4+, CD8+ and CD3+/CD8+ levels. Meanwhile, 
there were significant differences in SOD activities and rate of neutrophil phagocytosis between 
synergist groups and the control group. These results suggest that herbal immune synergist of Chinese 
traditional herbs prescription enhances the protective effect of classic swine fever vaccine. 

 
Key words: Immune synergist, Chinese medicinal herbs, classic swine fever vaccine. 

 
 
INTRODUCTION 

 
Many Chinese medicinal herbs are high efficacious immune 

synergist, including radix ginseng, astragalus, Szechwan 

asiabell root, poria mushroom, Chinese angelica and licorice 

root, which have been proven to possess immune-enhancement 

effects (Hu, 1997; Liu, 1998). These herbs contain plenty 

polysaccharides, saponins and flavones that are capable of 

modulating vertebrate cellular and humeral immunity, and 

disease resistance (Liu et al., 2006; Gao et al., 2000). Because 

the prescription is made up of pure Chinese medicinal herbs 

that are readily available, of low-cost and low side effects (Kong 

et al., 2004, 2006; Wang et al., 2005; Ung et al., 2007), it has 

become a research focus recently. However, most studies are 

centered on single herb; little attention has been paid to 

compound herbs. We have previously reported the effect of 

Chinese medicinal herbs synergist on chicken (Wang, et al., 

2006). Here, we described its synergistic effects on vaccination 

against classic swine fever.  
 
 

 
*Corresponding author. E-mail: aboluo2011888@163.com Tel: 
+86-0351-7561528. Fax: +86-0351-7561990. 

 
 
 
 

 
MATERIALS AND METHODS 
 
Animal selection and management 
 
Piglets (n = 60) were provided by Hongdong County Great Locust 
Tree Ecotech Limited, China. The feed was manufactured by Wuike 
Feedstuff Scientific Limited, China, according to the formula 
provided by the research group. Piglets were 50 to 60 days of age 
with body weight of 20 ± 2 kg. All animals were not previously 
exposed to any vaccine. Piglets were managed with the same 
standard of nutrition and procedure. Records and management 
were performed by designated personnel. 

 
Compound herbs immune synergist 
 
Ginseng (Panax ginseng C.A.Mey), milk vetch root (Astragalus 
membranaceus Bge var. mongholicus), Szechwan asiabell root 
(Codonopsis tangshen Oliv), poria mushroom (Poria cocos Schw. 
Wolf), Chinese angelica (Angelica sinensis Oliv. Diels), large head 
atractylodes rhozome (Atractylodes macrocephala Koidz), fructus 
amomi (Amomum villosum Lour), pericarpium citri reticulatae 
(Citrus reticulata Blanco), American ginseng (Panax quinquefolium 
L.), and licorice root (Glycyrrhiza uralensis Fisch) were used. The 
herbs were supplied by Taiyuan School of Chinese Traditional 
Medicine. 

http://www.sxagri.ac.cn/lijy/English/animalvet.htm
mailto:aboluo2011888@163.com


 
 
 

 
Table 1. Effects of herbal immune synergist on serum IgG content.  

 
 

Group 
 Time (day)  

 

 

10 20 30 40 
 

  
 

 A (1.0%) 606.25±43.04
b
 612.17±40.55

b
 618.46±34.95

b
 616.65±44.81

b
 

 

 B (1.5%) 617.25±43.04
a
 619.17±40.55

a
 628.66±34.95

a
 626.65±45.11

a
 

 

 C (2.0%) 618.02±42.34
a
 619.19±41.35

a
 629.12±33.85

a
 625.35±44.21

a
 

 

 D (control) 561.25±43.04
c
 598.17±40.55

c
 609.46±34.95

c
 607.65±44.81

c
 

 

 
Data presented are means ± SE of 15 replicates. For each column, means with the same letter are not significantly 
different (p < 0.05). 

 

 
Table 2. Effects of herbal immune synergist on serum IgM content. 

 

 
Group 

 Time (day)  
 

 

10 20 30 40  

  
 

 A (1.0%) 320.31±21.45
b
 329.65±27.01

b
 332.87±26.68

b
 330.60±25.36

b
 

 

 B (1.5%) 336.54±25.91
a
 339.51±24.91

a
 342.57±25.81

a
 340.55±24.92

a
 

 

 C (2.0%) 336.88±26.19
a
 339.89±26.21

a
 343.21±24.92

a
 41.33±23.71

a
 

 

 D (control) 309.51±24.42
c
 318.60±25.61

c
 322.58±26.11

c
 319.96±24.61

c
 

 

 
Data presented are means ± SE of 15 replicates. For each column, means with the same letter are not significantly 
different (p < 0.05). 

 

 

Vaccine and chemicals 
 
Vaccine was bought from Lanzhou Biological Pharmaceuticals  of  
Zhongmu Industry limited China. Sodium heparin, 
phytohemagglutinin (PHA), trypan blue, lipopolysaccharide and 3-  
(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide 
(MTT) were produced by Guangzhou Medical Industry Institute, 
China. Lymphocyte isolation solution was produced by Chinese 
Medical Academy Biotech Institute. Aluminum hydroxide gel was 
supplied by Longkel Biotech Pharmaceuticals, China. Calf serum, 
antibiotics, 0.8% NH4Cl solution, Wright’s dye and medium were 
prepared by the authors. 

 

Experimental design 
 
Randomized block design was used in this study. The 60 piglets 
were randomly assigned to 4 groups (n = 15 in each group). Piglets 
in the synergist group were treated with classic swine fever vaccine 
and herbal immune synergist. Starting from the first day of the 
experiment, piglets in the synergist group were fed, respectively 
with supplemented 1.0% (A group), 1.5% (B group) and 2.0% (C 
group) synergist prescription. On day 7, piglets were injected s.c. 
with vaccine, while piglets in the control group (n = 15) were treated 
with swine fever vaccine alone (D group). The whole experiment 
lasted for 60 days in which pretreatment lasted for 10 days and the 
treatment lasted for 50 days. Blood samples were collected from 
each group at day 10, 20, 30 and 40 after immune synergist 
treatment. 

 

Assay 

 
Content of serum IgG and IgM was determined by immune 
transmission turbidity. Peripheral T and B lymphocytes were 
measured with MTT colorimetric method. Red blood cell (RBC) and 
hemoglobin (Hb) were measured with general method. Serum GPT, 
GOT and SOD activity were measured with Automatic biochemical 
analyzer (TOSHIBA, Tokyo, Japan). Analysis on subpopulation of T 

 
 

 
lymphocytes was used by Flow Cytometry (Becton Dickinson, 
Rutherford, NJ, USA). Phagocytotic neutrophil assay was 
performed as follows: 
 

Phagocytotic neutrophil 
Phagocytosis rate = × 100%  

Total neutrophil 

 

Data analysis 
 
The data were subjected to one-way analyses of variance (ANOVAs) 

using the statistical package Minitab 15. Where required, Tukey multiple 

comparison tests were used following ANOVAs to identify significant 

differences between individual treatments. 
 
 
RESULTS 
 
Effects of herbal immune synergist on serum IgG and 
IgM contents 
 
Serum IgG and IgM contents are shown in Tables 1 and  
2. There were significant differences in serum IgG and 
IgM contents on day 10 between the synergist groups 
and control (P < 0.05). On day 30, serum IgG and IgM 
reached a peak, where average IgG levels in synergist 
groups were 2.54% higher than that in the control group 
(P < 0.05). Group C had a higher IgG content than group 
B, but no significant differences (P > 0.05). 
 
 
Effects of herbal immune synergist on peripheral T 
and B lymphocyte proliferation 

 

Groups B and C had  a higher B and T lymphocyte proli- 



 
 
 

 
Table 3. Effects of herbal immune synergist on peripheral B and T lymphocyte proliferation rate (OD).  

 

Group Lymphocyte 
 Time (day)  

 

10 20 30 40 
 

  
 

A (1.0%) 
T 0.68±0.32

b
 0.87±0.53

c
 1.07±0.61

b
 1.05±0.43

c
 

 

B 0.69±0.41
c
 0.88±0.56

b
 1.09±0.60

c
 1.08±0.46

c
 

 

 
 

B (1.5%) 

T 0.81±0.23
a
 0.98±0.49

b
 1.18±0.57

a
 1.13±0.41

b
 

 

B 0.79±0.21
b
 0.99±0.51

a
 1.17±0.58

b
 1.15±0.39

b
 

 

C (2.0%) 
T 0.82±0.35

a
 1.01±0.55

a
 1.19±0.61

a
 1.16±0.51

a
 

 

B 0.83±0.32
a
 1.00±0.60

a
 1.20±0.63

a
 1.18±0.50

a
 

 

 
 

D (control) 
T 0.60±0.25

c
 0.77±0.51

d
 0.99±0.52

c
 0.87±0.38

d
 

 

B 0.62±0.31
d
 0.78±0.48

c
 0.98±0.54

d
 0.89±0.37

d
 

 

 
 

       

 
Data presented are means ± SE of 15 replicates. For each column, means with the same letter are not significantly 
different (p < 0.05). 

 

 
Table 4. Effect of herbal immune synergist on T lymphocyte subpopulation on day 40.  

 

Group 
 Item (X ± SD)  

 

    
 

CD3
+
 CD4

+
 CD8

+
 CD3

+
 / CD8

+
 

 

 
 

      

A (1.0%) 45.38±0.32
b
 27.55±0.23

c
 21.96±0.35

c
 1.34±0.25

a
 

 

B (1.5%) 47.87±0.53
a
 29.76±0.49

b
 22.86±0.55

b
 1.35±0.51

a
 

 

C (2.0%) 47.88±0.61
a
 29.78±0.57

a
 22.89±0.61

a
 1.35±0.52

a
 

 

D (control) 44.75±0.43
c
 26.79±0.41

d
 21.92±0.52

d
 1.31±0.38

b
 

  
 

Data presented are means ± SE of 15 replicates. For each column, means with the same letter are not significantly 
different (p < 0.05). 

 

 

feration rate (Table 3). From day 10 to 30, proliferation 
rate showed an increasing tendency. On day 40, the 
lymphocyte proliferation decreased slightly. On day 30, B 
lymphocyte proliferation rates in groups B and C showed 
significant difference (P < 0.05), but T lymphocyte 
proliferation rates in group C were slightly higher than in 
group B, but there was no significant difference (P > 
0.05). 

 

Effect of herbal immune synergist on T lymphocyte 
subpopulation 
 
Table 4 shows the effect of traditional Chinese herbs 
prescription on subpopulation of T lymphocytes on day  
40. The ratio of T lymphocyte subpopulation in the 
synergist group was markedly increased and improved. 

 

Effects of herbal immune synergist on porcine red 
blood cell and Hb content 
 

Piglet red blood cell and hemoglobin contents are shown 
in Table 5. There were no significant differences between 

 
 

 

synergist groups and the control group (P > 0.05). 
 

 

Effects of herbal immune synergist on serum GPT, 
GOT and SOD activity 

 

Serum SOD activities in synergist groups were 
significantly different from that of the control group on 
days 30 and 40 (P < 0.05) (Table 6). GOT and GOT 
activities were similar between synergist groups and the 
control group (P > 0.05). 
 

 

Effects of herbal immune synergist on rate of 
neutrophil phagocytosis 
 

Rate of neutrophil phagocytosis showed a trend of 
increase in synergist groups, and reached a peak on day 
30 (Table 7). Rate of neutrophil phagocytosis of synergist 
groups was higher than that of the control group (P < 
0.05), but there was no significant difference between 
synergist groups B and C (P > 0.05). Rate of neutrophil 
phagocytosis of synergist groups decreased slightly on 



 
 
 

 
Table 5. Effects of herbal immune synergist on porcine red blood cell and Hb content.  

 

Group Item 
 Time (day)   

 

10 20 30 40 
 

 

   
 

A (1.0%) 

RBC 6.68±0.42
a
 6.78±0.33

a
 6.87±0.51

a
 6.75±0.53

a
 

 

Hb 12.09±0.51
a
 12.58±0.46

a
 12.89±0.50

a
 12.25±0.48

a
 

 

B (1.5%) 

RBC 6.81±0.33
a
 6.88±0.29

a
 6.97±0.47

a
 6.90±0.45

a
 

 

Hb 12.32±0.21
a
 12.99±0.51

a
 13.15±0.58

a
 12.96±0.39

a
 

 

C (2.0%) 

RBC 6.83±0.45
a
 6.92±0.56

a
 6.99±0.51

a
 6.95±0.46

a
 

 

Hb 12.43±0.42
a
 12.68±0.60

a
 13.18±0.82

a
 13.01±0.58

a
 

 

D (control) 

RBC 6.59±0.35
a
 6.63±0.46

a
 6.82±0.58

a
 6.80±0.58

a
 

 

Hb 11.62±0.61
a
 11.88±0.48

a
 12.38±0.46

a
 12.71±0.67

a
  

 

 
Data presented are means ± SE of 15 replicates. For each column, means with the same letter are not significantly different 
(p < 0.05). 

 

 
Table 6. Effects of herbal immune synergist on serum GPT, GOT and SOD activity.  
.   

Group Item 
 Time (day)  

 

10 20 30 40 
 

  
 

 GPT 35.68±0.42
a
 33.12±0.33

a
 31.87±0.51

a
 32.98±0.53

a
 

 

A (1.0%) GOT 36.09±0.51
a
 34.58±0.46

a
 32.89±0.50

a
 33. 05±0.48

a
 

 

 SOD 58.56±0.51
a
 98.55±0.46

a
 102.89±0.50

b
 98. 95±0.48

b
 

 

 GPT 35.51±0.33
a
 33.08±0.29

a
 30.27±0.47

a
 30.80±0.45

a
 

 

B (1.5%) GOT 36. 32±0.21
a
 32.99±0.51

a
 30.05±0.58

a
 30.96±0.39

a
 

 

 SOD 57. 82±0.21
a
 98.99±0.51

a
 123.15±0.58

a
 118.96±0.39

a
 

 

 GPT 35.88±0.45
a
 33.92±0.56

a
 31.39±0.51

a
 31.99±0.46

a
 

 

C (2.0%) GOT 36.43±0.42
a
 34.68±0.60

a
 30.18±0.82

a
 30.88±0.58

a
 

 

 SOD 57.43±0.42
a
 97.68±0.60

a
 123.18±0.82

a
 119.79±0.58

a
 

 

 GPT 35.59±0.35
a
 34.13±0.46

a
 33.82±0.58

a
 35.80±0.58

a
 

 

D (Control) GOT 34.62±0.61
a
 33.18±0.48

a
 32.48±0.46

a
 32.98±0.67

a
 

 

 SOD 57.62±0.61
a
 67.88±0.48

b
 78.38±0.46

c
 70.71±0.67

c
 

 

 
Data presented are means ± SE of 15 replicates. For each column, means with the same letter are not significantly different ( p < 
0.05). GPT, Glutamate-pyruvate transaminase; GOT, glutamate-oxaloacetate transaminase; SOD, superoxide dismutase. 

 

 

day 40, which was still higher than that of the control 
group (P < 0.05). 
 

 

DISCUSSION 

 

Animal immunity is closely related to disease resistance. 
When immune function decreases or certain killing 
mechanism is damaged, pathogens may invade animals, 
resulting to clinical symptoms. Up to now, vaccination is 
still the most effective preventing measure. Although, the 
strict immune program has been taken in the farms, some 
infectious diseases are still hard to be controlled 

 
 

 

Chinese medicinal herbs has been proven to be possess 
immune-enhancing properties (Xu et al., 2010, 2011). In 
addition, Chinese medicinal herbs may have many 
advantages including extensive availability, lower cost, 
reliable efficacy, decreased risk of side-effects and 
toxicity. Therefore, Chinese medicinal herbs or their 
ingredients have become a hotspot in recent years. The 
veterinary immune synergist prepared in this study 
selected herbs containing polysaccharides, flavones and 
saponins that are capable of activating immune system 
and stimulating lymphocytes (Jiang et al., 2010; Meng et 
al., 2011; Perera et al., 2010). In the study, our results 
demonstrate that supplementation of herbal immune 



 
 
 

 
Table 7. Effects of herbal immune synergist on rate of neutrophil phagocytosis.  
.   
 

Group 
  Time (day)  

 

 

10 20 30 40 
 

  
 

 A(1.0%) 0.068±0.21
b
 0.152±0.43

b
 0.216±0.51

b
 0.212±0.43

b
 

 

 B(1.5%) 0.074±0.23
a
 0.198±0.42

a
 0.276±0.53

a
 0.274±0.45

a
 

 

 C(2.0%) 0.075±0.22
a
 0.199±0.45

a
 0.276±0.47

a
 0.275±0.51

a
 

 

 D(blank) 0.057±0.24
c
 0.127±0.3 

c
 0.146±0.55

c
 0.181±0.43

c
 

 

 
Data presented are means ± SE of 15 replicates. For each column, means with the same letter are not significantly different 
(p < 0.05). 

 

 

synergist could increase serum IgG and IgM contents 
upon vaccination against classic swine fever, which is 
consistent with the report by Kong et al. (2004) and Hu et 
al. (2004). Herbal immune synergist promotes T and B 
lymphocyte proliferation and improves T lymphocyte 
subpopulation composition, which is consistent with the 
results reported by Chu et al. (2004). On the other hand, 
there were no significant differences for red blood cell and 
Hb content between synergist groups and the control 
group, and there were no significant differences between 
synergist groups and the control group in GPT and GOT. 
It indicates that these Chinese medicinal herbs are not 
toxic to the parenchymal of liver and heart. In addition, 
SOD activities and rate of neutrophil phagocytosis were 
markedly increased by herbal immune synergist. 
Although, 2.0% dose supplementation had a slightly 
better result than 1.5% dose, we recommend1.5% as the 
optimal dose.  

Vaccine is an important method of animal infectious 
diseases prevention and control, the synergy and 
mechanism between the vaccine and immune synergist 
needs to be studied further. In addition, immune synergist 
is not effective against all diseases, therefore it is 
important to continue to develop new immune synergist, 
and to strengthen its mechanism research. Chinese 
traditional medicinal herb is a potential candidate as a 
safe immune synergist for animal infectious diseases 
prevention and control. 
 

 

ACKNOWLEDGEMENT 

 

This work was supported by grants from Key 
Technologies Research and Development Program of 
Shanxi province (No. 983196). 
 

 
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