Full length articleDietary chitosan nanoparticles protect crayfish Procambarus clarkii against white spot syndrome virus (WSSV) infection
Introduction
Chitosan, derived by the deacetylation of chitin, is a natural polysaccharide composed of randomly distributed chains of β-(1-4) d-glucosamine and N-acetyl-d-glucosamine. Chitosan nanoparticles serve a better inhibitory activity against Candida albicans and the inhibitory effect was influenced by particle size [1]. Chitosan nanoparticles also could inhibit the growth of Escherichia coli, Salmonella choleraesuis, Salmonella typhimurium and Staphylococcus aureus [2]. Chitosan nanoparticles showed strong activity to inhibit the proliferation of human lymphoma cells through inducing apoptosis [3]. Chitosan nanoparticles have showed the antibacterial or anticancer activity in many studies. However, little research show the effect of chitosan nanoparticles on antiviral immunity of animals.
White spot syndrome virus (WSSV), which was first discovered in Taiwan in 1992, has caused mass mortalities and devastating production losses to shrimp farming over many areas [4], [5], [6]. The DNA genome of WSSV is about 300 kb, and WSSV may be the sole member of Nimaviridae, genus Whispovirus [7], [8]. WSSV is known to infect many crustacean species, including crayfish [9], [10]. Both farmed and wild Procambarus clarkii in Louisiana (USA) were natural hosts for WSSV [11], and P. clarkii has also been used for the study of antiviral immunity in shrimp [12]. In this study, we investigated the effect of chitosan nanoparticles protect P. clarkii against WSSV infection. We demonstrated that chitosan nanoparticles could improve the immuno-enzyme activities and enhance the antiviral defenses in crayfish.
Section snippets
Materials
Chitosan from Alaska snow crab shell was purchased from Qianguang biotechnology company (Tsingtao, China) and the degree of deacetylation is above 90%. The chitosan nanoparticles were prepared according to the previous report [13].
Characterizations of the nanoparticles
Particle size was measured using a Zetasizer Nano-ZS-90 (Malvern Instruments, UK). The analysis was performed at a scattering angle of 90° under 25 °C in triplicate.
Crayfish and WSSV stock
Crayfish P. clarkii, approximately 20 g and 8 cm each, were reared at 25 °C. They were kept in tanks
The particle size of chitosan nanoparticles
The size is essential characteristic parameters for a kind of nanoparticles. The data showed the size of the chitosan nanoparticles and most chitosan nanoparticles had a diameter between 10 nm and 20 nm (Fig. 1B). And the untreated chitosan particles had a diameter above 1000 nm (Fig. 1A). Using transmission electron microscopy, the morphology of chitosan and chitosan nanoparticles was shown about 1000 and 20 nm in diameter with spherical in shape and found to be agreement (Fig. 2).
Effects of chitosan nanoparticles on the mortality of WSSV-challenged crayfish
To evaluate
Discussion
Chitosan nanoparticles often were used to encapsulate DNA vaccine for oral delivery in shrimp [18], [19], fish [20] and other animals [21], [22], and a few reports showed that chitosan nanoparticles can enhance the immunogenicity of DNA vaccine [23]. The dietary chitosan nanoparticles supplementation may improve the growth performance and meat quality status of tilapia [24]. However, the effect of chitosan nanoparticles on protecting crustacean against virus infection did not be investigated
Acknowledgments
This work was financially supported by National Natural Science Foundation of China (31430089 and 31370050), the Major State Basic Research Development Program (2012CB114403).
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