Animal Bioscience and Biotechnology Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, USDA, Beltsville, MD 20705, USA.
Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Poult Sci. 2022 Apr;101(4):101731. doi: 10.1016/j.psj.2022.101731. Epub 2022 Jan 15.
The in vitro antimicrobial activity of sophorolipids (SLs) against Eimeria maxima and Clostridium perfringens, and the in vivo effects of SLs on growth performance and gut health in necrotic enteritis (NE)-afflicted broiler chickens were studied. To test the direct killing effects of SLs on enteric pathogens, 2.5 × 10 freshly prepared sporozoites of each Eimeria acervulina, E. maxima, and E. tenella were placed in each well of a 96-well plate, and the vegetative stage of Clostridium perfringens was prepared at 1 × 10 cfu/well. Four different SLs (C18:1 lactonic diacetyled SL [SL1], C18:1 deacetyled SL [SL2], C18:1 monoacetyled SL [SL3], and C18:1 diacetyled SL [SL4]), and 2 anticoccidial chemical controls, decoquinate and monensin, were evaluated at 3 dose levels (125 µg/mL, 250 µg/mL, and 500 µg/mL). Samples were incubated at 41°C for 3 h, and microbial survival ratios were measured by using a cell counter to quantify the number of live microbes stained by fluorescent dye. A total of 336 (0-day-old) male commercial broiler chickens were used to assess the effects of SLs in vivo. Chickens were randomly allocated to 6 treatment groups (7 chickens per cage, 8 cages per treatment) as follows: a control group which received a basal diet (CON), a negative control group (NC) which received a basal diet and NE challenge, and 4 SL treatment groups with NE (NC+SL1, NC+SL2, NC+SL3, and NC+SL4). The inclusion rates of SLs in each group were 200 mg/kg of feed. NE-induced chickens were orally infected with E. maxima (10,000 oocysts/chicken) on d 14, followed by C. perfringens (1 × 10 cfu/chicken) on d 19. Disease parameters measured included gut lesion scores, intestinal cytokine production, and level of tight junction protein expression. Data were analyzed using a Mixed Model (PROC MIXED) in SAS. In vitro (Experiment 1), all SLs dose-dependently decreased (P < 0.001) the viability of the three species of Eimeria sporozoites and C. perfringens. In vivo (Experiment 2), dietary SLs increased (P < 0.001) body weight and average daily gain of broiler chickens infected with NE. Dietary SL1 and SL4s increased (P < 0.05) feed conversion ratio compared to NC. Furthermore, SL1 and SL4 decreased (P < 0.05) gut lesion scores in combination with increased expression of IL1β, IL8, TNFSF15, and IL10 genes (P < 0.05) in NE-afflicted chickens. Overall, dietary SLs promoted growth performance, intestinal immune responses, and intestinal barrier integrity of NE-afflicted, young broiler chickens.
研究了槐糖脂(SLs)对巨型艾美耳球虫和产气荚膜梭菌的体外抗菌活性,以及 SLs 对坏死性肠炎(NE)感染肉鸡生长性能和肠道健康的体内影响。为了测试 SLs 对肠道病原体的直接杀伤作用,将每种柔嫩艾美耳球虫、巨型艾美耳球虫和堆型艾美耳球虫的 2.5×10 个新鲜制备的孢子虫放入 96 孔板的每个孔中,将产气荚膜梭菌的营养阶段制备成 1×10 cfu/孔。评估了 4 种不同的 SLs(C18:1 内酯二乙酰化 SL [SL1]、C18:1 去乙酰化 SL [SL2]、C18:1 单乙酰化 SL [SL3]和 C18:1 二乙酰化 SL [SL4])和 2 种抗球虫化学对照药癸氧喹酯和莫能菌素,在 3 个剂量水平(125 µg/mL、250 µg/mL 和 500 µg/mL)下进行评估。样品在 41°C 下孵育 3 小时,并用细胞计数器测量微生物存活率,通过荧光染料染色定量活微生物的数量来测量微生物的存活比例。共使用 336 只(0 日龄)雄性商业肉鸡评估 SLs 的体内效果。肉鸡随机分配到 6 个处理组(每组 7 只鸡,每组 8 个笼),如下所示:接受基础日粮的对照组(CON)、接受基础日粮和 NE 挑战的阴性对照组(NC)和 4 个接受 NE 的 SL 处理组(NC+SL1、NC+SL2、NC+SL3 和 NC+SL4)。每组 SL 的添加率为 200 mg/kg 饲料。NE 诱导的鸡在第 14 天口服感染巨型艾美耳球虫(10,000 个卵囊/鸡),然后在第 19 天口服感染产气荚膜梭菌(1×10 cfu/鸡)。测量的疾病参数包括肠道病变评分、肠道细胞因子产生和紧密连接蛋白表达水平。数据使用 SAS 中的混合模型(PROC MIXED)进行分析。在体外(实验 1),所有 SLs 剂量依赖性地降低(P<0.001)三种艾美耳球虫孢子虫和产气荚膜梭菌的活力。在体内(实验 2),日粮 SLs 增加(P<0.001)感染 NE 的肉鸡的体重和平均日增重。与 NC 相比,日粮 SL1 和 SL4 增加(P<0.05)饲料转化率。此外,SL1 和 SL4 降低(P<0.05)了 NE 感染鸡的肠道病变评分,并增加了 IL1β、IL8、TNFSF15 和 IL10 基因的表达(P<0.05)。总的来说,日粮 SLs 促进了感染 NE 的年轻肉鸡的生长性能、肠道免疫反应和肠道屏障完整性。