Thymol Improves Barrier Function and Attenuates Inflammatory Responses in Porcine Intestinal Epithelial Cells during Lipopolysaccharide (LPS)-Induced Inflammation.

It is well-known that essential oil thymol exhibits antibacterial activity. The protective effects of thymol on pig intestine during inflammation is yet to be investigated. In this study, an in vitro lipopolysaccharide (LPS)-induced inflammation model using IPEC-J2 cells was established. Cells were pretreated with thymol for 1 h and then exposed to LPS for various assays. Interleukin 8 (IL-8) secretion, the mRNA abundance of cytokines, reactive oxygen species (ROS), nutrient transporters, and tight junction proteins was measured. The results showed that LPS stimulation increased IL-8 secretion, ROS production, and tumor necrosis factor alpha (TNF-α) mRNA abundance ( P < 0.05), but the mRNA abundance of sodium-dependent glucose transporter 1 (SGLT1), excitatory amino acid transporter 1 (EAAC1), and H+/peptide cotransporter 1 (PepT1) were decreased ( P < 0.05). Thymol blocked ROS production ( P < 0.05) and tended to decrease the production of LPS-induced IL-8 secretion ( P = 0.0766). The mRNA abundance of IL-8 and TNF-α was reduced by thymol pretreatment ( P < 0.05), but thymol did not improve the gene expression of nutrient transporters ( P > 0.05). The transepithelial electrical resistance (TEER) was reduced and cell permeability increased by LPS treatment ( P < 0.05), but these effects were attenuated by thymol ( P < 0.05). Moreover, thymol increased zonula occludens-1 (ZO-1) and actin staining in the cells. However, the mRNA abundance of ZO-1 and occludin-3 was not affected by either LPS or thymol treatments. These results indicated that thymol enhances barrier function and reduce ROS production and pro-inflammatory cytokine gene expression in the epithelial cells during inflammation. The regulation of barrier function by thymol and LPS may be at post-transcriptional or post-translational levels.

Essential oils as alternatives to antibiotics in swine production.

This review article summarizes the efficacy, feasibility and potential mechanisms of the application of essential oils as antibiotic alternatives in swine production. Although there are numerous studies demonstrating that essential oils have several properties, such as antimicrobial, antioxidative and anti-inflammatory effects, feed palatability enhancement and improvement in gut growth and health, there is still a need of further investigations to elucidate the mechanisms underlying their functions. In the past, the results has been inconsistent in both laboratory and field studies because of the varied product compositions, dosages, purities and growing stages and conditions of animals. The minimal inhibitory concentration (MIC) of essential oils needed for killing enteric pathogens may not ensure the optimal feed intake and the essential oils inclusion cost may be too high in swine production. With the lipophilic and volatile nature of essential oils, there is a challenge in effective delivery of essential oils within pig gut and this challenge can partially be resolved by microencapsulation and nanotechnology. The effects of essential oils on inflammation, oxidative stress, microbiome, gut chemosensing and bacterial quorum sensing (QS) have led to better production performance of animals fed essential oils in a number of studies. It has been demonstrated that essential oils have good potential as antibiotic alternatives in feeds for swine production. The combination of different essential oils and other compounds (synergistic effect) such as organic acids seems to be a promising approach to improve the efficacy and safety of essential oils in applications. High-throughput systems technologies have been developed recently, which will allow us to dissect the mechanisms underlying the functions of essential oils and facilitate the use of essential oils in swine production.

Development of Novel Microparticles for Effective Delivery of Thymol and Lauric Acid to Pig Intestinal Tract.

Antibiotics have been widely supplemented in feeds at subtherapeutic concentrations to prevent postweaning diarrhea and increase the overall productivity of pigs. However, the emergence of antimicrobial-resistant bacteria worldwide has made it urgent to minimize the use of in-feed antibiotics. The development of promising alternatives to in-feed antibiotics is crucial for maintaining the sustainability of swine production. Both medium-chain fatty acids (MCFA) and essential oils exhibit great potential to postweaning diarrhea; however, their direct inclusion has compromised efficacy because of several factors including low stability, poor palatability, and low availability in the lower gut. Therefore, the objective of this study was to develop a formulation of microparticles to deliver a model of essential oil (thymol) and MCFA (lauric acid). The composite microparticles were produced by the incorporation of starch and alginate through a melt-granulation process. The release of thymol and lauric acid from the microparticles was in vitro determined using simulated salivary fluid (SSF), simulated gastric fluid (SGF), and simulated intestinal fluid (SIF), consecutively. The microparticles prepared with 2% alginate solution displayed a slow release of thymol and lauric acid in the SSF (21.2 ± 2.3%; 36 ± 1.1%), SGF (73.7 ± 6.9%; 54.8 ± 1.7%), and SIF (99.1 ± 1.2%; 99.1 ± 0.6%), respectively, whereas, the microparticles without alginate showed a rapid release of thymol and lauric acid from the SSF (79.9 ± 11.8%; 84.9 ± 9.4%), SGF (92.5 ± 3.5%; 75.8 ± 5.9%), and SIF (93.3 ± 9.4%; 93.3 ± 4.6%), respectively. The thymol and lauric acid in the developed microparticles with or without alginate both exhibited excellent stabilities (>90%) during being stored at 4 °C for 12 weeks and after being stored at room temperature for 2 weeks. These results evidenced that the approach developed in the present study could be potentially employed to deliver thymol and lauric acid to the lower gut of pigs, although further in vivo investigations are necessary to validate the efficacy of the microparticles.