Oral administration of <i>Allium sativum</i> extract protects against infectious bursal disease in chickens

doi:10.15302/J-FASE-2015080

Front. Agr. Sci. Eng.

2015, Vol. 2

Issue (4) : 318-326 https://doi.org/10.15302/J-FASE-2015080

RESEARCH ARTICLE

Oral administration of Allium sativum extract protects against infectious bursal disease in chickens

Sufen ZHAO,Yuanyuan JIA,Weiwei ZHANG,Lili WANG,Yunfei MA(

),Kedao TENG(

)

College of Veterinary Medicine, China Agricultural University, Beijing 100193, China

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Abstract

Garlic (Allium sativum, Liliaceae) has been safely used for more than 5000 years, and research on garlic extract is rapidly increasing because of its multiple biological functions. The in vivo effects of oral administration of garlic mixture (GM, water-soluble extract) on infectious bursal disease virus (IBDV)-infected specific pathogen free male white leghorn chicken were examined through histopathological, immunohistochemical, and Western blot analyses, and enzyme-linked immunosorbent assay. The results confirmed the protective effects of oral administration of 5 mg·kg⁻¹ BW GM (Group GM1) on bursal lesions after IBDV infection. In particular, protein expression of IBDV in the bursa decreased in Group GM1, indicating that GM administration decreased IBDV replication in the bursa. Furthermore, immunoglobulin M- and A-bearing B lymphocytes significantly increased 7 days post infection in bursae in Group GM1 (P<0.01), suggesting that the oral administration of 5 mg·kg⁻¹ GM offers moderate protection against B cell destruction after IBDV infection. During infection, the concentration of bursal interferon gamma (IFN-g) increased and peaked in Group GM1 earlier than in Group T (IBDV-exposed), demonstrating that GM administration prompted the production of IFN-g to protect against IBDV infection.

Keywords garlic infectious bursal disease virus (IBDV) antiviral effect IgM-bearing B lymphocyte

Corresponding Author(s): Yunfei MA,Kedao TENG

Just Accepted Date: 11 December 2015 Online First Date: 31 December 2015 Issue Date: 19 January 2016

Cite this article:

Sufen ZHAO,Yuanyuan JIA,Weiwei ZHANG, et al. Oral administration of Allium sativum extract protects against infectious bursal disease in chickens[J]. Front. Agr. Sci. Eng. , 2015, 2(4): 318-326.

URL:

https://academic.hep.com.cn/fase/EN/10.15302/J-FASE-2015080
https://academic.hep.com.cn/fase/EN/Y2015/V2/I4/318

Fig.1 Effect of GM on the bursal weight index of IBDV-infected chickens. Mean bursal indexes of Groups GM1 and GM2 at 1 to 7 and 2 to 7 dpi, respectively, were higher than the mean bursal index of Group T. Among them, bursal indexes of Groups GM1 and GM2 at 3 and 3 to 4 dpi, respectively, were remarkably higher (P<0.05). Bursal indexes of Group T were significantly lower than those of Group C at 4 to 7 dpi (P<0.05). However, a significant decrease occurred in Groups GM1 and GM2 at 5 to 7 and 7 dpi, respectively (P<0.05). Data were given as mean±SD. Data simultaneously marked with different letters indicate significant differences (P<0.05).

Fig.2 Effect of GM on the bursal structure changes of IBDV-infected chickens. At 7 dpi, a few injured bursal lymphoid follicles were still observed in Group T, and local infiltration of lymphocytes (LIL) and fibrillation were investigated in the interfollicular areas (a, b). Many LIL and a high number of original bursal lymphoid follicles were detected with orderly cortical and medullary epithelial cells and many lymphocytes in the cortex and medulla in Group GM1 (c, d). The most serious congestion, edema, hemorrhage and heterophil accumulation were observed in Group GM2, and a high number of degenerated tissues accumulated in the bursal tube, with few lymphoid follicles (e, f). Arrows indicate fibrillation. Arrowheads indicate LIL.

Fig.3 Effect of GM on the B lymphocyte changes of IBDV-infected chickens. A few IgM⁺B lymphocytes were scattered in the tissues where lesions involuted in Group T, and LIL were IgM negative (a, b). A high number of IgM⁺B lymphocytes were detected in the cortex and medulla in Group GM1 (c, d). Fewer IgM⁺B lymphocytes were observed in the bursal epibiotic area (EA) of Group GM2 (e, f). At 7 dpi, many IgA⁺B lymphocytes were observed in the cortex and medulla in Group GM1, and LIL were IgA negative (g, h). Few IgA⁺B lymphocytes were observed in Groups T (i, j) and GM2 (k, l). At 7 dpi, IgM⁺B lymphocytes were enumerated in five fields/bursa/chicken, and a significantly higher number of IgM⁺B lymphocytes in Group GM1 and the epibiotic area of Group GM2 was observed than in Group T (P <0.01) (m). The number of IgA⁺B lymphocytes in Group GM1 was significantly higher than that in Group T at 7 dpi (P<0.01) (n). Solid arrows and arrowheads indicate scattered IgM immunoreactivities and IgM⁺B lymphocytes, respectively. Feint arrows and arrowheads indicate scattered IgA immunoreactivities and IgA⁺B lymphocytes, respectively. Data are given as mean±SD. **, P<0.01 between Groups GM1 and T; ^##, P<0.01 between Groups GM2 and T.

Fig.4 Effect of GM on the expression of bursal viral antigens of IBDV-infected chickens. At 7 dpi, many VP2 immunoreactivities were concentrated in the medulla of injured bursal lymphoid follicles, and some diffusely distributed in the cortex and interfollicular areas in Group T (a). A few VP2 immunoreactivities exhibited diffuse distribution in the plicae in Group GM1 (b). The strongest VP2 immunoreactivities were observed in Group GM2, with most accumulated in degenerated tissues (c). Western blot analysis (d, e) confirmed the low VP2 expression in GM1.

Fig.5 Effect of GM on IFN-g concentrations in the bursal homogenate of IBDV-infected chickens. After infection, bursal IFN-g concentrations increased in the chickens. Among them, IFN-g concentrations in Groups T and GM1 initially increased and subsequently decreased, with a significant increase at 4 and 5 dpi in Group T and 2 to 4 dpi in Group GM1 (P<0.05). A significantly higher bursal IFN-g concentration was observed in Group GM2 at 4 and 7 dpi (P<0.05). The IFN-g concentration peaked at 3 dpi in Group GM1, 2 days earlier than that in Groups T and GM2. Data are given as mean±SD. Data simultaneously marked with different letters indicate significant differences (P<0.05).

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