Feeding-Regime-Dependent Intestinal Response of Rainbow Trout after Administration of a Novel Probiotic Feed.

Intensive fish farming is associated with a high level of stress, causing immunosuppression. Immunomodulators of natural origin, such as probiotics or phytoadditives, represent a promising alternative for increasing the immune function of fish. In this study, we tested the autochthonous trout probiotic strain L. plantarum R2 in a newly developed, low-cost application form ensuring the rapid revitalization of bacteria. We tested continuous and cyclic feeding regimes with regard to their effect on the intestinal immune response and microbiota of rainbow trout. We found that during the continuous application of probiotic feed, the immune system adapts to the immunomodulator and there is no substantial stimulation of the intestinal immune response. During the cyclic treatment, after a 3-week break in probiotic feeding and the reintroduction of probiotics, there was a significant stimulation of the gene expression of molecules associated with both cellular and humoral immunity (CD8, TGF-ß, IL8, TLR9), without affecting the gene expression for IL1 and TNF-α. We can conclude that, in aquaculture, this probiotic feed can be used with a continuous application, which does not cause excessive immunostimulation, or with a cyclic application, which provides the opportunity to stimulate the immunity of trout, for example, in periods of stress.

Silybin Showed Higher Cytotoxic, Antiproliferative, and Anti-Inflammatory Activities in the CaCo Cancer Cell Line while Retaining Viability and Proliferation in Normal Intestinal IPEC-1 Cells.

The anticancer potential of silymarin is well known, including its anti-inflammatory as well as antiproliferative effect mediated by influencing the cell cycle, suppression of apoptosis, and inhibition of cell-survival kinases. However, less is known about silybin, the main component of the silymarin complex, where studies indicate its dual effect on the proliferation and immune response of various cell types in a dose-dependent manner. Moreover, there is a lack of studies comparing the effect of silybin on the same type of healthy and tumor cells, especially intestinal ones. Therefore, our study aimed to investigate the concentration-dependent effect of silybin on the normal intestinal porcine epithelial cell line-1 (IPEC-1) and the human epithelial colorectal adenocarcinoma cell line (CaCo-2). The metabolic viability, cell cycle, mitochondrial membrane potential, apoptosis, and the relative gene expression for pro- and anti-inflammatory cytokines were monitored in cells treated with silybin. Silybin stimulates metabolic viability as well as proliferation in IPEC-1 cells, protects the mitochondrial membrane, and thus exerts a cytoprotective effect, and has only a minimal effect on the gene expression of pro-inflammatory cytokines but significantly increases the expression of anti-inflammatory TGF-ß. In contrast, it inhibits metabolic viability in tumor intestinal CaCo-2 cells, has an antiproliferative effect accompanied by increased apoptosis, and significantly reduces the expression of genes for pro-inflammatory interleukins as well as TGF-ß. The antiproliferative and anti-inflammatory effect of silybin on tumor intestinal cells without a negative effect on healthy cells is a prerequisite for its potential use in the adjuvant therapy of colon cancer; however, further studies are necessary.

The adverse effects of synthetic acaricide tau-fluvalinate (tech.) on winter adult honey bees.

Currently several pyrethroids (e.g., flumethrin and tau-fluvalinate) are used in apiculture worldwide as acaricides/miticides. The long half-lives of pyrethroids in synthetic acaricides applied to hive matrices, may adversely affect the health of bee colony. The potentially adverse effects of synthetic acaricide/miticide tau-fluvalinate (tech.) on winter honeybees were assessed in this study (OECD 245 2017). No dose-dependent mortality in in vitro reared winter honeybees was observed after chronic oral 10-day exposure to syrup (50% w/v) spiked with a maximum concentration of 750 µg a.i./kg diet and its 1/10 concentration. The No Observed Effect Concentration is ≥ 750 µg a.i./kg diet. Tau-fluvalinate testing for the sublethal effects on bee immune system showed up-regulated gene expression encoding abaecin, lysozyme, and defensin in both tested groups, however the expression of hymenoptaecin gene was reduced. Moreover, tau-fluvalinate significantly induced levels of DNA damage in exposed bees, which can result in adverse genotoxic effect.

Effect of autochthonous lactobacilli on immunologically important molecules of rainbow trout after bacterial infection studied on intestinal primoculture.

Nowadays, the aquaculture industry is one of the fastest growing industries. Intensive aquaculture has a negative impact on fish health. Probiotic bacteria are often used due to beneficial effect to health of host, e.i. decrease of diseases outbreaks, immunomodulatory effect or better utilization of feed. The aim of this work was to study the influence of probiotic bacteria on the immune response of trout intestinal cells in primoculture infected with pathogenic bacteria. In the experiment, we tested the effect of pre-treatment of intestinal cells with an autochthonous strain of Lactobacillus plantarum R2 Biocenol™ (CCM 8674) following infection with the most serious salmonid pathogens - Aeromonas salmonicida subsp. salmonicida (CCM 1307) and Yersinia ruckeri (CCM 6093). Tested probiotic strain reduced inflammation after A. salmonicida infection through decreased expression of pro-inflammatory cytokines and increased expression of anti-inflammatory cytokines. In contrast, after infection with Y. ruckeri, which causes immunosuppression, the probiotic strain stimulated immunity by up-regulation of expression of proinflammatory cytokines and suppressed the expression of anti-inflammatory cytokines. These results are a prerequisite for the immunomodulatory potential of the strain, but its action must be confirmed in subsequent in vivo experiments.