Extraction of lipopolysaccharide from fish-derived Aeromonas hydrophila and their immunostimulatory effect on Qihe crucian carp
Purified lipopolysaccharide (LPS) was first obtained from fish-derived Aeromonas hydrophila (A. hydrophila) by hot phenol-water combined with enzymatic hydrolysis. Subsequently, different concentrations of LPS (0, 0.2, 0.4, 0.8, and 1.2 mg/mL) were injected intraperitoneally into Qihe crucian carp (...
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KeAi Communications Co., Ltd.
2025-09-01
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| Series: | Aquaculture and Fisheries |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2468550X24000558 |
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| author | Xinxin Liu Yiran Wang Junli Wang Xiao Yan Liping Yang Chaobin Qin Guoxing Nie |
| author_facet | Xinxin Liu Yiran Wang Junli Wang Xiao Yan Liping Yang Chaobin Qin Guoxing Nie |
| author_sort | Xinxin Liu |
| collection | DOAJ |
| description | Purified lipopolysaccharide (LPS) was first obtained from fish-derived Aeromonas hydrophila (A. hydrophila) by hot phenol-water combined with enzymatic hydrolysis. Subsequently, different concentrations of LPS (0, 0.2, 0.4, 0.8, and 1.2 mg/mL) were injected intraperitoneally into Qihe crucian carp (Carassius auratus var. Qihe) with a dose of 3 μL/g body weight to evaluated their immunostimulatory activity by detecting the expression of immune-related genes in the spleen, head kidney, and hepatopancreas. The results showed that the average extraction rate of LPS was about 1.063% based on the precipitation weight of wet bacteria, of which the polysaccharide content was 9.39%. The silver staining band of LPS after SDS-PAGE electrophoresis showed a ladder distribution typically characterized by smooth-type LPS. Protein and nucleic acid electrophoresis results showed very low amounts of residual protein and nucleic acid, and high-performance liquid chromatography (HPLC) results showed that the purity of the extracted A. hydrophila LPS was similar to that of high-purity commercial LPS. Pro-inflammatory cytokine genes such as IL-1, IL-8, TNF-α, and IL-6 were more sensitive to LPS stimulation than anti-inflammatory factors such as IL-10 and TGF-β. After 2 h of injection, their expression levels in the spleen, head kidney, and hepatopancreas were significantly increased (P < 0.05), with the 0.8 and 1.2 mg/mL injection groups showing the most significant increase. The gene expression of IL-10 and TGF-β showed slight fluctuations in response to A. hydrophila LPS stimulation (P < 0.05). In conclusion, high-purity LPS was extracted from A. hydrophila derived from fish by modified hot phenol-water method, it demonstrating that LPS product could significantly regulate the innate immunity of Qihe crucian carp, with the most effective vaccination dose being 2 mg/kg/BW. These findings provide useful information for future research on endotoxin vaccines. |
| format | Article |
| id | doaj-art-4041d17f22e54816bc6baf80cf4077b8 |
| institution | Kabale University |
| issn | 2468-550X |
| language | English |
| publishDate | 2025-09-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Aquaculture and Fisheries |
| spelling | doaj-art-4041d17f22e54816bc6baf80cf4077b82025-08-22T04:57:00ZengKeAi Communications Co., Ltd.Aquaculture and Fisheries2468-550X2025-09-0110584385110.1016/j.aaf.2024.04.003Extraction of lipopolysaccharide from fish-derived Aeromonas hydrophila and their immunostimulatory effect on Qihe crucian carpXinxin Liu0Yiran Wang1Junli Wang2Xiao Yan3Liping Yang4Chaobin Qin5Guoxing Nie6College of Fisheries, Henan Normal University, No. 46 Jianshe Road, Xinxiang, 453007, ChinaCollege of Life Sciences, Henan Normal University, No. 46 Jianshe Road, Xinxiang, 453007, ChinaCollege of Life Sciences, Henan Normal University, No. 46 Jianshe Road, Xinxiang, 453007, China; Corresponding author.College of Fisheries, Henan Normal University, No. 46 Jianshe Road, Xinxiang, 453007, ChinaCollege of Fisheries, Henan Normal University, No. 46 Jianshe Road, Xinxiang, 453007, ChinaCollege of Fisheries, Henan Normal University, No. 46 Jianshe Road, Xinxiang, 453007, ChinaCollege of Fisheries, Henan Normal University, No. 46 Jianshe Road, Xinxiang, 453007, China; Corresponding author.Purified lipopolysaccharide (LPS) was first obtained from fish-derived Aeromonas hydrophila (A. hydrophila) by hot phenol-water combined with enzymatic hydrolysis. Subsequently, different concentrations of LPS (0, 0.2, 0.4, 0.8, and 1.2 mg/mL) were injected intraperitoneally into Qihe crucian carp (Carassius auratus var. Qihe) with a dose of 3 μL/g body weight to evaluated their immunostimulatory activity by detecting the expression of immune-related genes in the spleen, head kidney, and hepatopancreas. The results showed that the average extraction rate of LPS was about 1.063% based on the precipitation weight of wet bacteria, of which the polysaccharide content was 9.39%. The silver staining band of LPS after SDS-PAGE electrophoresis showed a ladder distribution typically characterized by smooth-type LPS. Protein and nucleic acid electrophoresis results showed very low amounts of residual protein and nucleic acid, and high-performance liquid chromatography (HPLC) results showed that the purity of the extracted A. hydrophila LPS was similar to that of high-purity commercial LPS. Pro-inflammatory cytokine genes such as IL-1, IL-8, TNF-α, and IL-6 were more sensitive to LPS stimulation than anti-inflammatory factors such as IL-10 and TGF-β. After 2 h of injection, their expression levels in the spleen, head kidney, and hepatopancreas were significantly increased (P < 0.05), with the 0.8 and 1.2 mg/mL injection groups showing the most significant increase. The gene expression of IL-10 and TGF-β showed slight fluctuations in response to A. hydrophila LPS stimulation (P < 0.05). In conclusion, high-purity LPS was extracted from A. hydrophila derived from fish by modified hot phenol-water method, it demonstrating that LPS product could significantly regulate the innate immunity of Qihe crucian carp, with the most effective vaccination dose being 2 mg/kg/BW. These findings provide useful information for future research on endotoxin vaccines.http://www.sciencedirect.com/science/article/pii/S2468550X24000558Aeromonas hydrophilaLipopolysaccharideExtraction and purificationImmune-related genes |
| spellingShingle | Xinxin Liu Yiran Wang Junli Wang Xiao Yan Liping Yang Chaobin Qin Guoxing Nie Extraction of lipopolysaccharide from fish-derived Aeromonas hydrophila and their immunostimulatory effect on Qihe crucian carp Aquaculture and Fisheries Aeromonas hydrophila Lipopolysaccharide Extraction and purification Immune-related genes |
| title | Extraction of lipopolysaccharide from fish-derived Aeromonas hydrophila and their immunostimulatory effect on Qihe crucian carp |
| title_full | Extraction of lipopolysaccharide from fish-derived Aeromonas hydrophila and their immunostimulatory effect on Qihe crucian carp |
| title_fullStr | Extraction of lipopolysaccharide from fish-derived Aeromonas hydrophila and their immunostimulatory effect on Qihe crucian carp |
| title_full_unstemmed | Extraction of lipopolysaccharide from fish-derived Aeromonas hydrophila and their immunostimulatory effect on Qihe crucian carp |
| title_short | Extraction of lipopolysaccharide from fish-derived Aeromonas hydrophila and their immunostimulatory effect on Qihe crucian carp |
| title_sort | extraction of lipopolysaccharide from fish derived aeromonas hydrophila and their immunostimulatory effect on qihe crucian carp |
| topic | Aeromonas hydrophila Lipopolysaccharide Extraction and purification Immune-related genes |
| url | http://www.sciencedirect.com/science/article/pii/S2468550X24000558 |
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