Leyva-Diaz Anaisa A, Hernandez-Patlan Daniel, Solis-Cruz Bruno, Adhikari Bishnu, Kwon Young Min, Latorre Juan D, Hernandez-Velasco Xochitl, Fuente-Martinez Benjamin, Hargis Billy M, Lopez-Arellano Raquel, Tellez-Isaias Guillermo
Departamento de Medicina y Zootecnia de Aves, Facultad de Medicina Veterinaria y Zootecnia, UNAM, 04510, Ciudad de Mexico, Mexico.
Laboratorio 5: LEDEFAR, Unidad de Investigacion Multidisciplinaria, Facultad de Estudios Superiores (FES) Cuautitlan, Universidad Nacional Autonoma de Mexico (UNAM), 54714, Cuautitlan Izcalli, Mexico.
J Anim Sci Biotechnol. 2021 Feb 5;12(1):23. doi: 10.1186/s40104-021-00545-7.
Interest in the use of natural feed additives as an alternative to antimicrobials in the poultry industry has increased in recent years because of the risk of bacterial resistance. One of the most studied groups are polyphenolic compounds, given their advantages over other types of additives and their easy potentiation of effects when complexes are formed with metal ions. Therefore, the objective of the present study was to evaluate the impact of dietary supplementation of copper acetate (CA), curcumin (CR), and their combination (CA-CR) against Salmonella Typhimurium colonization, intestinal permeability, and cecal microbiota composition in broiler chickens through a laboratory Salmonella infection model. S. Typhimurium recovery was determined on day 10 post-challenge by isolating Salmonella in homogenates of the right cecal tonsil (12 chickens per group) on Xylose Lysine Tergitol-4 (XLT-4) with novobiocin and nalidixic acid. Intestinal integrity was indirectly determined by the fluorometric measurement of fluorescein isothiocyanate dextran (FITC-d) in serum samples from blood obtained on d 10 post-S. Typhimurium challenge. Finally, microbiota analysis was performed using the content of the left caecal tonsil of 5 chickens per group by sequencing V4 region of 16S rRNA gene.
The results showed that in two independent studies, all experimental treatments were able to significantly reduce the S. Typhimurium colonization in cecal tonsils (CT, P < 0.0001) compared to the positive control (PC) group. However, only CA-CR was the most effective treatment in reducing S. Typhimurium counts in both independent studies. Furthermore, the serum fluorescein isothiocyanate dextran (FITC-d) concentration in chickens treated with CR was significantly lower when compared to PC (P = 0.0084), which is related to a decrease in intestinal permeability and therefore intestinal integrity. The effect of dietary treatments in reducing Salmonella was further supported by the analysis of 16S rRNA gene sequences using Linear discriminant analysis effect size (LEfSe) since Salmonella was significantly enriched in PC group (LDA score > 2.0 and P < 0.05) compared to other groups. In addition, Coprobacillus, Eubacterium, and Clostridium were significantly higher in the PC group compared to other treatment groups. On the contrary, Fecalibacterium and Enterococcus in CR, unknown genus of Erysipelotrichaceae at CA-CR, and unknown genus of Lachnospiraceae at CA were significantly more abundant respectively.
CR treatment was the most effective treatment to reduce S. Typhimurium intestinal colonization and maintain better intestinal homeostasis which might be achieved through modulation of cecal microbiota.
近年来,由于细菌耐药性的风险,家禽业对使用天然饲料添加剂替代抗菌药物的兴趣有所增加。多酚类化合物是研究最多的群体之一,因为它们比其他类型的添加剂具有优势,并且与金属离子形成复合物时易于增强效果。因此,本研究的目的是通过实验室沙门氏菌感染模型,评估日粮中添加醋酸铜(CA)、姜黄素(CR)及其组合(CA-CR)对肉鸡鼠伤寒沙门氏菌定植、肠道通透性和盲肠微生物群组成的影响。在攻毒后第10天,通过在含有新生霉素和萘啶酸的木糖赖氨酸胆盐-4(XLT-4)培养基上从右盲肠扁桃体匀浆(每组12只鸡)中分离沙门氏菌,来测定鼠伤寒沙门氏菌的回收率。在鼠伤寒沙门氏菌攻毒后第10天采集的血液血清样本中,通过荧光法测量异硫氰酸荧光素葡聚糖(FITC-d)间接测定肠道完整性。最后,通过对每组5只鸡的左盲肠扁桃体内容物进行16S rRNA基因V4区测序来进行微生物群分析。
结果表明,在两项独立研究中,与阳性对照组(PC)相比,所有实验处理均能显著降低盲肠扁桃体中鼠伤寒沙门氏菌的定植(CT,P < 0.0001)。然而,在两项独立研究中,只有CA-CR是降低鼠伤寒沙门氏菌数量最有效的处理方法。此外,与PC组相比,用CR处理的鸡血清中异硫氰酸荧光素葡聚糖(FITC-d)浓度显著降低(P = 0.0084),这与肠道通透性降低以及肠道完整性有关。使用线性判别分析效应大小(LEfSe)对16S rRNA基因序列进行分析,进一步支持了日粮处理对减少沙门氏菌的作用,因为与其他组相比,PC组中沙门氏菌显著富集(LDA分数> 2.0且P < 0.05)。此外,与其他处理组相比,PC组中的粪杆菌属、真杆菌属和梭菌属显著更高。相反,CR组中的粪杆菌属和肠球菌属、CA-CR组中丹毒丝菌科的未知属以及CA组中毛螺菌科的未知属分别显著更丰富。
CR处理是减少鼠伤寒沙门氏菌肠道定植并维持更好的肠道内环境稳定的最有效处理方法,这可能是通过调节盲肠微生物群实现的。