Scientia Agricultura Sinica ›› 2026, Vol. 59 ›› Issue (15): 3452-3464.doi: 10.3864/j.issn.0578-1752.2026.15.016

• ANIMAL SCIENCE·VETERINARY SCIENCE • Previous Articles    

Establishment of an Infection Model in Pigs for Chinese Clinical Strains of Lawsonia intracellularis

XIE Rui1(), GU ZhengQuan1, LIU SiWen1, LUO YaJuan1, CUI YuCong1, HUA Lin1, SONG WenBo2, HUANG Chao2, TANG XiBiao2, XU GaoYuan2, CHEN HuanChun1, WU Bin1(), PENG Zhong1()   

  1. 1 College of Veterinary Medicine Huazhong Agricultural University/Hubei Hongshan Laboratory/The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070
    2 Wuhan Keqian Biology Co., Ltd., Wuhan 430073
  • Received:2026-01-29 Accepted:2026-06-04 Online:2026-08-01 Published:2026-08-03
  • Contact: WU Bin, PENG Zhong

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Abstract:

【Objective】This study aimed to establish a stable porcine proliferative enteropathy (PPE) model using Chinese isolates of Lawsonia intracellularis (LI) and clinical infected intestinal tissue homogenates from different sources, construct a systematic evaluation index system covering fecal bacterial shedding, fecal score, average daily gain, feed conversion ratio, serum antibody level and histopathological changes, and identify the optimal challenge dose and preferred strain, so as to provide the standardized model and data support for the research on LI pathogenic mechanism, vaccine efficacy evaluation and prevention and control strategies.【Method】Twenty-eight-day-old weaned piglets were selected and divided into a negative control group and multiple infection groups. In Experiment 1, the pathogenicity of four LI tissue homogenates (JX703, GX301, HN312, and LI-HuB23) was compared at the dose of 1×109 LI per piglet. In Experiment 2, JX703 tissue homogenate with the strongest comprehensive pathogenicity and the in vitro cultured LI-HuB23 isolate were selected to orally inoculate 28-day-old piglets at gradient doses (1010 LI per pig, 108 LI per pig, and 106 LI per pig). The piglets were monitored for 28 days after inoculation, during which fecal bacterial shedding, fecal score, average daily gain, feed conversion ratio, serum antibody level and histopathological changes were observed and recorded.【Result】In Experiment 1, typical clinical symptoms and pathological manifestations of PPE were successfully reproduced in all infection groups inoculated with tissue homogenates of different LI strains. LI was detectable in feces from day 4 post-inoculation, and bacterial shedding persisted until day 28 post-inoculation. Diarrhea occurred in the infection groups from day 4 post-inoculation, with the most severe symptoms observed from day 19 to 21 post-inoculation. All infection groups showed lower average daily gain and higher feed conversion ratio compared with the negative control group. Serum antibody monitoring showed that seroconversion occurred from day 14 after the establishment of infection and lasted until day 28 post-inoculation. Pathological examination revealed thickened intestinal mucosal folds in the infection groups; hematoxylin-eosin (HE) staining showed intestinal crypt hyperplasia and decreased goblet cells, and LI was detected in the cytoplasm of intestinal crypt epithelial cells by immunohistochemical staining (IHC). Overall, JX703 tissue homogenate induced the most prominent symptoms with the strongest virulence. In Experiment 2, the results of gradient dose test showed that inoculation with LI-HuB23 in 106 LI per piglet was sufficient to establish effective infection, accompanied by fecal bacterial shedding, diarrhea, and corresponding pathological changes. The severity of symptoms increased with the elevation of infection dose, and the 1010 LI per pig group exhibited the most typical patterns in terms of peak fecal scores, pathological scores, and reductions in average daily gain.【Conclusion】A PPE infection model was successfully established using Chinese LI isolates and field-derived infected tissue homogenates in this study. The selected evaluation indexes had an important reference value for related research, including fecal bacterial shedding, fecal score, average daily gain, feed conversion ratio, serum antibody level and histopathological changes. The infection model established with an inoculation dose of 1010 LI per pig presented more typical clinical and pathological characteristics of PPE.

Key words: porcine proliferative enteropathy, Lawsonia intracellularis, swine, infection model, challenge dose

Fig. 1

Monitoring of indicators in pigs after infection with different tissue homogenates of LI A: Fecal LI content at different time points post-infection; B: Fecal consistency scores of pigs in each group at different time points post-infection; C: Average daily gain (ADG) of pigs in each group across different infection stages; D: Representative fecal morphology of diseased pigs: (a) Watery feces, (b) Soft feces. Note: Negative control represents pigs orally administered with sterile SPG; JX703, GX301, HN312, and LI-HuB23 represent pigs challenged with different LI tissue homogenates, respectively. Data are shown as mean±SD. *P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001 versus the negative control group at the matched time point"

Table 1

Feed conversion ratio of experimental pigs in each group (kg·kg-1)"

时间 Time 阴性对照 Negative control JX703 GX301 HN312 LI-HuB23
0-28d 2.66±0.39 3.65±1.14 3.47±0.56 3.91±1.30 3.13±0.47

Fig. 2

Pathological scores of intestinal tissues in experimental pigs Negative control represents pigs orally administered with sterile SPG; JX703, GX301, HN312, and LI-HuB23 represent pigs challenged with different LI tissue homogenates, respectively. *P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001 versus the negative control group at the matched time point"

Fig. 3

Gross and histopathological changes of intestinal tissues in experimental pigs Yellow arrows in H.E staining indicate crypt hyperplasia and goblet cell reduction; red arrows in IHC staining indicate LI colonization. Scale bar =100 μm. Negative Control represents pigs orally administered with sterile SPG; JX703, GX301, HN312, and LI-HuB23 represent pigs challenged with different LI tissue homogenates, respectively"

Fig. 4

Preparation of LI antibody detection plate Nuclei were stained with DAPI (blue), and LI was labeled with specific green fluorescence"

Fig. 5

Detection results of serum anti-LI antibodies in experimental pigs Nuclei were stained with DAPI (blue), and green fluorescence represents LI-specific fluorescent signal; +, positive for serum LI antibody; -, negative for serum LI antibody. Negative control represents pigs orally administered with sterile SPG; JX703, GX301, HN312, and LI-HuB23 represent pigs challenged with different LI tissue homogenates, respectively"

Fig. 6

Monitoring of indicators in pigs after infection with different doses of the LI-HuB23 isolate A: Fecal LI content at different time points post-infection; B: Fecal consistency scores of pigs in each group at different time points post-infection; C: Average daily gain (ADG) of pigs in each group across different infection stages; D: Representative fecal morphology of diseased pigs: (a) Soft feces, (b) Watery feces. Note: Negative Control represents pigs orally administered with sterile SPG; LI-HuB23-1010 LI, LI-HuB23-108 LI, and LI-HuB23-106 LI represent pigs orally challenged with LI-HuB23 isolate at the dose of 1×1010, 1×108, and 1×106 LI per pig, respectively; JX703-1010 LI, JX703-108 LI, and JX703-106 LI represent pigs orally challenged with JX703 tissue homogenate at the dose of 1×1010, 1×108, and 1×106 LI per pig, respectively. Data are shown as mean±SD. **** P<0.0001 versus the negative control group at the matched time point"

Table 2

Feed conversion ratio of pigs in each group"

时间
Time		 阴性对照
Negative control		 LI-HuB23
1010 LI		 LI-HuB23
108 LI		 LI-HuB23
106 LI		 JX703
1010 LI		 JX703
108 LI		 JX703
106 LI
0-28d 3.09±0.57 4.03±1.02 3.69±0.43 3.64±1.29 4.44±1.09 4.58±2.83 3.19±0.55

Fig. 7

Pathological scores of intestinal tissues in experimental pigs Negative control represents pigs orally administered with sterile SPG; LI-HuB23-1010 LI, LI-HuB23-108 LI, and LI-HuB23-106 LI represent pigs orally challenged with LI-HuB23 isolate at the dose of 1×1010, 1×108, and 1×106 LI per pig, respectively; JX703-1010 LI, JX703-108 LI, and JX703-106 LI represent pigs orally challenged with JX703 tissue homogenate at the dose of 1×1010, 1×108, and 1×106 LI per pig, respectively. *P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001 versus the negative control group at the matched time point"

Fig. 8

Gross and histopathological changes of intestinal tissues in experimental pigs Yellow arrows in H.E staining indicate crypt hyperplasia and goblet cell reduction; red arrows in IHC staining indicate LI colonization. Scale bar =100 μm. Negative Control represents pigs orally administered with sterile SPG; LI-HuB23-1010 LI, LI-HuB23-108 LI, and LI-HuB23-106 LI represent pigs orally challenged with LI-HuB23 isolate at the dose of 1×1010, 1×108, and 1×106 LI per pig, respectively; JX703-1010 LI, JX703-108 LI, and JX703-106 LI represent pigs orally challenged with JX703 tissue homogenate at the dose of 1×1010, 1×108, and 1×106 LI per pig, respectively"

Fig. 9

Detection results of serum anti-LI antibodies in experimental pigs Nuclei were stained with DAPI (blue), and green fluorescence represents LI-specific fluorescent signal; +, positive for serum LI antibody; -, negative for serum LI antibody. Negative control represents pigs orally administered with sterile SPG; LI-HuB23-1010 LI, LI-HuB23-108 LI, and LI-HuB23-106 LI represent pigs orally challenged with LI-HuB23 isolate at the dose of 1×1010, 1×108, and 1×106 LI per pig, respectively; JX703-1010 LI, JX703-108 LI, and JX703-106 LI represent pigs orally challenged with JX703 tissue homogenate at the dose of 1×1010, 1×108, and 1×106 LI per pig, respectively"

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