Assessment of Fecal Microflora Changes in Pigs Supplemented with Herbal Residue and Prebiotic

Abstract

Antibiotic usage in animals as a growth promoter is considered as public health issue due to its negative impact on consumer health and environment. The present study aimed to evaluate effectiveness of herbal residue (ginger, Zingiber officinale, dried rhizome powder) and prebiotic (inulin) as an alternative to antibiotics by comparing fecal microflora composition using terminal restriction fragment length polymorphism. The grower pigs were offered feed containing antibiotic (tetracycline), ginger and inulin separately and un-supplemented group served as control. The study revealed significant changes in the microbial abundance based on operational taxonomic units (OTUs) among the groups. Presumptive identification of organisms was established based on the fragment length of OTUs generated with three restriction enzymes (MspI, Sau3AI and BsuRI). The abundance of OTUs representing Bacteroides intestinalis, Eubacterium oxidoreducens, Selonomonas sp., Methylobacterium sp. and Denitrobacter sp. was found significantly greater in inulin supplemented pigs. Similarly, the abundance of OTUs representing Bacteroides intestinalis, Selonomonas sp., and Phascolarcobacterium faecium was found significantly greater in ginger supplemented pigs. In contrast, the abundance of OTUs representing pathogenic microorganisms Atopostipes suicloacalis and Bartonella quintana str. Toulouse was significantly reduced in ginger and inulin supplemented pigs. The OTUs were found to be clustered under two major phylotypes; ginger-inulin and control-tetracycline. Additionally, the abundance of OTUs was similar in ginger and inulin supplemented pigs. The results suggest the potential of ginger and prebioticsto replace antibiotics in the diet of grower pig.

Fig 1. Histogram showing abundance response of 16s rRNA OTUs generated by MspI (A), Sau3AI (B) and BsuRI (C) restriction enzymes.

The 16s rRNA gene was amplified from fecal microflora of control or tetracycline, ginger and inulin supplemented pigs.

Fig 2. Scatter plot matrix showing the relative response of the 16s rRNA gene OTUs generated by MspI (A), Sau3AI (B) and BsuRI (C) restriction enzymes from pig fecal samples.

MPAC-Mean peak area of control pigs, MPAT—Mean peak area of tetracycline supplemented pigs, MPAG- Mean peak area of ginger supplemented pigs, MPAI- Mean peak area of inulin supplemented pigs.

Fig 3. Non-metric multi dimensional analysis of OTU abundance showing relationship among the individuals of four treatment groups.

The OTUs were generated from 16s rRNA gene amplified from pig fecal microflora of control or tetracycline, ginger and inulin supplemented groups. A: OTUs generated with MspI, B: OTUs generated with Sau3AI, C: OTUs generated BsuRI.