Montana Kaimin, March 5, 1981

Student newspaper of the University of Montana, Missoula.https://scholarworks.umt.edu/studentnewspaper/8304/thumbnail.jp

Morphometry of the distal intestine.

<p>Box-and-whiskerplots displaying the transformed relative length of the PCNA positive regions in the crypts of the distal intestine on the y-axis and the six feeding groups on the x-axis. Boxes mark the interquartile range with the black bar marking the median and whiskers extending to the interquartile range. Data points from the six different groups are plotted with a jitter in the x-direction for better visualisation. Groups with different letters on the upper x-axis are significantly different (multiple comparisons of means, Tukey, ).</p

Correlation of differentially expressed genes.

<p>Gene expression changes in the distal intestine of the study groups: CV, CU, KM, SC and SBM, each relative to the negative control group (FM) were correlated against each other. The <b>upper triangle</b> shows the scatter plots and Pearson correlations (<i>r</i>) of the for 3,791 genes that showed DE in at least one of the study groups. The <b>diagonal</b> displays the total number of DE genes for each study group, while the number of genes that are commonly DE between two groups are shown in the <b>lower triangle</b>.</p

<i>Candida utilis</i> and <i>Chlorella vulgaris</i> Counteract Intestinal Inflammation in Atlantic Salmon (<i>Salmo salar</i> L.)

<div><p>Intestinal inflammation, caused by impaired intestinal homeostasis, is a serious condition in both animals and humans. The use of conventional extracted soybean meal (SBM) in diets for Atlantic salmon and several other fish species is known to induce enteropathy in the distal intestine, a condition often referred to as SBM induced enteropathy (SBMIE). In the present study, we investigated the potential of different microbial ingredients to alleviate SBMIE in Atlantic salmon, as a model of feed-induced inflammation. The dietary treatments consisted of a negative control based on fish meal (FM), a positive control based on 20% SBM, and four experimental diets combining 20% SBM with either one of the three yeasts <i>Candida utilis</i> (CU), <i>Kluyveromyces marxianus</i> (KM), <i>Saccharomyces cerevisiae</i> (SC) or the microalgae <i>Chlorella vulgaris</i> (CV). Histopathological examination of the distal intestine showed that all fish fed the SC or SBM diets developed characteristic signs of SBMIE, while those fed the FM, CV or CU diets showed a healthy intestine. Fish fed the KM diet showed intermediate signs of SBMIE. Corroborating results were obtained when measuring the relative length of PCNA positive cells in the crypts of the distal intestine. Gene set enrichment analysis revealed decreased expression of amino acid, fat and drug metabolism pathways as well as increased expression of the pathways for NOD-like receptor signalling and chemokine signalling in both the SC and SBM groups while CV and CU were similar to FM and KM was intermediate. Gene expression of antimicrobial peptides was reduced in the groups showing SBMIE. The characterisation of microbial communities using PCR-DGGE showed a relative increased abundance of <i>Firmicutes</i> bacteria in fish fed the SC or SBM diets. Overall, our results show that both CU and CV were highly effective to counteract SBMIE, while KM had less effect and SC had no functional effects.</p></div

Validation of microarray results by qRT-PCR.

<p>LogFCs from the microarray experiment are plotted on the <i>x-axis</i>, while logFCs from the qRT-PCR are plotted on the <i>y-axis</i>. qRT-PCR data was normalised to an index of the genes EF1 and GAPDH. Expression levels of five different genes were compared. For each gene the logFCs for all five contrasts were calculated and plotted (as listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0083213#pone.0083213.s009" target="_blank">Table S3</a>). The dotted line represents the linear regression line. Pearson correlation and the corresponding <i>p-value</i> are displayed in the plot.</p

Ingredients and chemical composition of the experimental diets.

<p>FM: fish meal; SBM: soybean meal; CV: <i>Chlorella vulgaris</i>; CU: <i>Candida utilis</i>; KM: <i>Kluyveromyces marxianus</i>; SC: <i>Saccharomyces cerevisiae</i>.</p><p> Norse-LT 94, Nordsildmel, Egersund, Norway.</p><p> Denofa, Fredrikstad, Norway.</p><p> Rousselot SAS, Courbevoie, France.</p><p> Lyckeby Culinar AB, Fjlkinge, Sweden.</p><p> NorSalmOil, Norsildmel, Egersund, Norway.</p><p> Vitamin and mineral premix provided (per kg diet): all-trans retinyl acetate, 860 ; cholecalciferol, 37.5 ; D,L--tocopherol acetate, 200 ; menadione, 10 ; thiamin, 15 ; riboflavin, 25 ; nicotinic acid, 75 ; pantothenic acid, 30 ; pyridoxine, 15 ; folic acid, 5 ; cyanocobalamin, 20 ; ascorbyl monophosphate, 125 ; biotin, 0.25 ; Ca, 1.1 ; ZnSO, 296 ; MnSO, 41 ; , 13 ; , 2.6 ; CaI, 3.5 ; astaxanthin, 175 .</p><p> Synergy Natural Products Pty Ltd, Sydney, Australia.</p><p> Borregaard ASA, Sarpsborg, Norway.</p><p> YBD-Yeast Brewers, Sigma-Aldrich, St. Louis, USA.</p

Regulation of tryptophan metabolism.

<p>Heat map showing the mean gene expression of the 18 genes in the KEGG pathway tryptophan metabolism, per group relative to the FM group. An asterisks(*) labelling the group indicates significant difference between the respective group and FM. <i>Aldehyde dehydrogenase family 9 member A1-A</i> (A9A1A); <i>Probable arylformamidase</i> (AFMID); <i>Kynurenine/alpha-aminoadipate aminotransferase</i> (AADAT); <i>Aldehyde dehydrogenase, mitochondrial</i> (ALDH2); <i>Aromatic-L-amino-acid decarboxylase</i> (DDC); <i>Cytochrome P450 1A1</i> (CP1A1); <i>Kynureninase</i> (KYNU); <i>2-amino-3-carboxymuconate-6-semialdehyde decarboxylase</i> (ACMSD); <i>Aldehyde oxidase</i> (ADO); <i>Peroxisomal bifunctional enzyme</i> (ECHP); <i>Glutaryl-CoA dehydrogenase</i> (GCDH); <i>Indoleamine 2,3-dioxygenase 2</i> (I23O2); <i>Kynurenine 3-monooxygenase</i> (KMO); <i>Catalase</i> (CATA); <i>Angiotensin-converting enzyme 2</i> (ACE2); <i>Fatty aldehyde dehydrogenase</i>(AL3A2); <i>Tryptophan 5-hydroxylase 1</i> (TPH1); <i>Acetyl-CoA acetyltransferase</i> (THIC).</p

Identification of dominant bacteria in distal intestine.

<p><b>A:</b> Dendrogram showing the similarities between DGGE profiles of PCR-amplified fragments of the 16S rRNA gene (V6–V8 region). The dendrogram was created using the Jaccard similarity coefficient as distance measure and Ward's clustering algorithm. <b>B:</b> Relative bacterial abundance in the distal intestine of the six different feeding groups. Abundance for a given bacterial species was calculated using the DGGE band intensities. <i>NSB*</i>  =  non sequenced DGGE bands.</p

Histopathological changes in the distal intestine.

<p>The histopathology scores from the different feeding groups are plotted as stacked bars. Each stacked bar shows the histopathology scores of 15 fish (x-axis) that were sampled per diet. Histology scores were given for the four categories: <b>A</b>: <i>lamina propria</i>; <b>B</b>: Epithelium; <b>C</b>: Atrophy; <b>D</b>: Oedema. Groups with different letters on the upper x-axis are significantly different (multiple comparisons of mean ranks, Nemenyi-Dunn, ).</p

Dietary regulation of KEGG pathways.

<p>Heat map showing pathway enrichment scores for the 109 KEGG pathways that were significantly affected by the diet (ANOVA, adjusted <i>p-value </i>). The pathways are grouped by function. Histology scores for <i>lamina propria</i>, epithelium and atrophy are colour coded and shown on top of the heat map.</p