Department of Animal Science, Iowa State University, Ames, IA.
Department of Veterinary Diagnostic and Production Animal Medicine, Iowa State University, Ames, IA.
J Anim Sci. 2020 May 1;98(5). doi: 10.1093/jas/skaa113.
The objective of this experiment was to investigate the impact of an F18 enterotoxigenic Escherichia coli (ETEC) challenge on growth performance, aspects of intestinal function, and selected immune responses of piglets, as well as to evaluate potential protective effects of direct-fed microbial (DFM) blends. Seventy-two weaned piglets (6.4 ± 0.2 kg body weight [BW]; 21 d of age) were assigned to one of four treatments: 1) NC: Nonchallenged (n = 10), 2) positive challenged control (PC): F18 ETEC-challenged (n = 10), 3) PC + DFM1 (n = 8; three strains of Bacillus amyloliquefaciens; 7.5 × 105 colony-forming units [cfu]/g), or 4) PC + DFM2 (n=8; 2 strains of B. amyloliquefaciens and one strain of Bacillus subtilis; 1.5 × 105 cfu/g). Feed intake and BW were recorded on day 0, 7, and 17. Pigs were sham-infected either with 6 mL phosphate-buffered saline or inoculated with 6 mL F18 ETEC (1.9 × 109 cfu/mL) on day 7 (0 d postinoculation [dpi]). All ETEC-challenged pigs were confirmed to be genetically susceptible to F18. Pigs had ad libitum access to feed and water throughout the 17-d trial. Fecal scores were visually ranked and rectal temperatures were recorded daily. To evaluate ETEC shedding, fecal swabs were collected on dpi 0, 1, 2, 3, 5, 7, and 10. Blood samples were collected on dpi 0, 1, 2, 4, 7, and 10. Ileal tissues were collected at necropsy on dpi 10. All challenged treatments had lower final BW, decreased average daily gain (ADG), and average daily feed intake (ADFI) during the 10-d postchallenge period (P < 0.01). The DFM2 treatment increased E. coli shedding on dpi 2 and decreased iton dpi 7 (P < 0.05) compared with the PC. Rectal temperature decreased across all challenged treatments (P < 0.01). Ileal mRNA abundance of occludin (OCLN) and zonula occludens-1 (ZO-1) decreased in PC and DFM1 compared with NC (P < 0.05). Pigs fed DFM2 had intermediate ileal mRNA abundance of OCLN and increased ZO-1 mRNA compared with pigs in PC (P < 0.05). Interleukin 8 (IL-8) increased in the plasma of PC and DFM2 on dpi 2 compared with NC (P < 0.05). Mucosal IL-8 increased in PC compared with NC (P < 0.05). All challenged treatments tended to have elevated tumor necrosis factor-α (TNF-α) mRNA abundance compared with NC (P < 0.10). Challenged pigs had reduced secretory immunoglobulin A and villus height compared with NC pigs (P < 0.05). The impact of an ETEC challenge on intestinal function and the immune system has been revealed, information critical to developing improved treatment regimes.
本实验的目的是研究 F18 肠产毒性大肠杆菌(ETEC)攻毒对仔猪生长性能、肠道功能和部分免疫反应的影响,并评估直接饲喂微生物(DFM)混合物的潜在保护作用。将 72 头断奶仔猪(6.4±0.2 公斤体重[BW];约 21 日龄)分为以下 4 种处理之一:1)NC:未攻毒(n=10),2)阳性攻毒对照组(PC):F18 ETEC 攻毒(n=10),3)PC+DFM1(n=8;3 株解淀粉芽孢杆菌;7.5×105 菌落形成单位[cfu]/g),或 4)PC+DFM2(n=8;2 株解淀粉芽孢杆菌和 1 株枯草芽孢杆菌;1.5×105 cfu/g)。在 0、7 和 17 天记录采食量和 BW。第 7 天(攻毒后 0 天[dpi]),猪用 6 mL 磷酸盐缓冲盐水或 6 mL F18 ETEC(~1.9×109 cfu/mL)进行假感染或攻毒。所有 ETEC 攻毒猪均被证实对 F18 具有遗传易感性。试验期间,所有仔猪均自由采食和饮水。攻毒后每天观察粪便评分和直肠温度。为评估 ETEC 脱落情况,于 dpi 0、1、2、3、5、7 和 10 采集粪便拭子。于 dpi 0、1、2、4、7 和 10 采集血液样本。于 dpi 10 剖检时采集回肠组织。与未攻毒组相比,所有攻毒组的终 BW、平均日增重(ADG)和平均日采食量(ADFI)在攻毒后 10 天内均较低(P<0.01)。与 PC 相比,DFM2 处理组在 dpi 2 时增加了大肠杆菌的脱落量,在 dpi 7 时减少了大肠杆菌的脱落量(P<0.05)。所有攻毒组的直肠温度均下降(P<0.01)。与 NC 相比,PC 和 DFM1 组回肠黏膜 occludin(OCLN)和 zonula occludens-1(ZO-1)的 mRNA 丰度降低(P<0.05)。与 PC 相比,DFM2 组仔猪回肠 OCLN 的 mRNA 丰度中等,ZO-1 的 mRNA 丰度增加(P<0.05)。攻毒后 2 天,PC 和 DFM2 组仔猪血浆中白细胞介素 8(IL-8)增加(P<0.05)。与 NC 相比,PC 组回肠黏膜 IL-8 增加(P<0.05)。与 NC 相比,所有攻毒组的肿瘤坏死因子-α(TNF-α)mRNA 丰度均有升高趋势(P<0.10)。攻毒组仔猪的分泌型免疫球蛋白 A 和绒毛高度均低于 NC 组(P<0.05)。本研究揭示了 ETEC 攻毒对仔猪肠道功能和免疫系统的影响,这对开发改进的治疗方案具有重要意义。