Administration of probiotics affects Artemia franciscana metanauplii intestinal ultrastructure and offers resistance against a Photobacterium damselae ssp. piscicida induced oxidative stress response

Maria Touraki , Anna Chanou , Vasiliki Mavridou , Vasiliki Tsertseli , Maria Tsiridi , Emmanuel Panteris
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Abstract

The effects of Photobacterium damselae ssp. piscicida (Phdp) on immune responses and intestinal ultrastructure of Artemia franciscana following infection and their amelioration by the probiotic bacteria Bacillus subtilis, Lactobacillus plantarum and Lactococcus lactis were evaluated. Pathogen growth inhibition in coculture with each probiotic and its virulence against Artemia were confirmed with an LC50 of 105 CFU mL−1. Phdp administration to Artemia at sublethal levels resulted in depletion of superoxide dismutase, glutathione reductase, glutathione transferase and phenoloxidase activities, extensive lipid peroxidation and reduced survival. Following a combined administration of each probiotic and the pathogen, enzyme activities and survival were significantly higher, while lipid peroxidation was reduced, compared to the infected group with no probiotic treatment (P < 0.05). The transmission electron microscopy study revealed that pathogen infection resulted in disarranged and fragmented microvilli, formation of empty or pathogen containing cytoplasmic vacuoles and damaged mitochondria. In the probiotic-treated and Phdp-infected series, intestinal cells showed normal appearance, except for the presence of pathogen-containing vacuoles and highly ordered but laterally stacked microvilli. The results of the present study indicate that Phdp induces cell death through an oxidative stress response and probiotics enhance Artemia immune responses to protect it against the Phdp induced damage.

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益生菌的施用影响了叶蒿(Artemia franciscana metanauplii)的肠道超微结构,并提供了对一种光细菌damselae ssp的抗性。双鱼诱导氧化应激反应
紫荆光杆菌对植物生长的影响。研究了piscicida (Phdp)对Artemia franciscana感染后的免疫反应和肠道超微结构的影响,以及枯草芽孢杆菌、植物乳杆菌和乳酸乳球菌对其的改善作用。各益生菌共培养对病原菌生长有抑制作用,其对蒿的毒力LC50为105 CFU mL−1。以亚致死水平给药的青蒿导致超氧化物歧化酶、谷胱甘肽还原酶、谷胱甘肽转移酶和酚氧化酶活性的消耗,广泛的脂质过氧化和生存降低。与未接受益生菌治疗的感染组相比,在联合使用每种益生菌和病原体后,酶活性和存活率显著提高,同时脂质过氧化降低(P <0.05)。透射电镜观察发现,病原菌感染导致微绒毛紊乱、破碎,细胞质空泡或含病原菌形成,线粒体受损。在益生菌处理和ph感染系列中,肠细胞外观正常,除了存在含病原体的液泡和高度有序但侧向堆叠的微绒毛。本研究结果表明,Phdp通过氧化应激反应诱导细胞死亡,益生菌增强青蒿免疫反应,保护其免受Phdp诱导的损伤。
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