Arkadios Dimitroglou

Microbial manipulations to improve fish health and production--a Mediterranean perspective.

The interactions between the endogenous gut microbiota and the fish host are integral in mediating the development, maintenance and effective functionality of the intestinal mucosa and gut associated lymphoid tissues (GALTs). These microbial populations also provide a level of protection against pathogenic visitors to the gastrointestinal (GI) tract and aid host digestive function via the production of exogenous digestive enzymes and vitamins. Manipulation of these endogenous populations may provide an alternative method to antibiotics to control disease and promote health management. Applications of probiotics for Mediterranean teleosts can stimulate immune responses, enhance growth performance, feed utilisation, digestive enzyme activities, antioxidant enzyme activities, gene expression, disease resistance, larval survival, gut morphology, modulate GI microbiota and mediate stress responses. Although considerably less information is available regarding prebiotic applications for Mediterranean teleosts, prebiotics also offer benefits with regards to improving immune status and fish production. Despite the promising potential benefits demonstrated in current literature, obtaining consistent and reliable results is often difficult due to our incomplete understanding of indigenous fish GI microbiota and their subsequent host interactions which mediate and drive both localised and systemic host immunological responses. Additionally, the probiotic and prebiotic (biotics) mechanisms which mediate host benefits at the mucosal interface are poorly understood. Future studies focused on these interactions utilising gnotobiotic techniques should provide a better understanding of how to extract the full potential of biotic applications to promote immune function of Mediterranean teleosts.

Dietary mannan oligosaccharide supplementation modulates intestinal microbial ecology and improves gut morphology of rainbow trout, Oncorhynchus mykiss (Walbaum).

A study was conducted to investigate the effect of mannan oligosaccharide (MOS) on the gut microbiota and intestinal morphology of rainbow trout under commercial farming conditions. Juvenile (mean initial BW 38.2 +/- 1.7 g) and subadult (111.7 +/- 11.6 g) trout were fed 2 dietary treatments for 111 and 58 d, respectively. The control treatment consisted of a standard commercial diet, and the MOS treatment consisted of the control diet supplemented with 0.2% MOS. Morphology of the anterior and the posterior intestine was examined with light and electron microscopy. Light microscopy demonstrated increased gut absorptive surface area in the subadult MOS group. Additionally, electron microscopy revealed an increase in microvilli length and density in the subadult MOS group compared with the control (P < 0.05). However, no significant improvements were detected in the juvenile group. Culture-based evaluation of the intestinal microbiota showed that MOS significantly reduced (P < 0.05) the viable intestinal bacterial populations (by approximately 2 log scales in all cases). Levels of Aeromonas/Vibrio spp. were significantly decreased (P < 0.05) in the juvenile MOS group (9% of the total microbiota) compared with the juvenile control group (37%). Additionally, analysis of microbial communities was conducted using denaturing gradient gel electrophoresis of PCR-amplified 16S rDNA. The denaturing gradient gel electrophoresis fingerprinting revealed an alteration of bacterial populations; analysis of similarity, similarity percentages, and nonmetric multidimensional scaling analysis showed that MOS reduced species richness and increased similarity of bacterial populations found within the subadult and juvenile groups. The current study shows that MOS modulates intestinal microbial communities, which subsequently improve gut morphology and epithelial brush border.

Soybean meal alters autochthonous microbial populations, microvilli morphology and compromises intestinal enterocyte integrity of rainbow trout, Oncorhynchus mykiss (Walbaum).

Abstract Rainbow trout were fed either a diet containing fishmeal (FM) as the crude protein source or a diet containing 50% replacement with soybean meal (SBM) for 16 weeks. An enteritis-like effect was observed in the SBM group; villi, enterocytes and microvilli were noticeably damaged compared with the FM group. The posterior intestine microvilli of SBM-fed fish were significantly shorter and the anterior intestine microvilli significantly less dense than the FM-fed fish. Electron microscopy confirmed the presence of autochthonous bacterial populations associated with microvilli of both fish groups. Reduced density of microvilli consequently led to increased exposure of enterocyte tight junctions, which combined with necrotic enterocytes is likely to diminish the protective barrier of the intestinal epithelium. No significant differences in total viable counts of culturable microbial populations were found between the groups in any of the intestinal regions. A total of 1500 isolates were tentatively placed into groups or genera, according to standard methods. Subsequent partial 16S rRNA sequencing revealed species that have not been identified from the rainbow trout intestine previously. Compared with the FM group levels of Psychrobacter spp. and yeast were considerably higher in the SBM group; a reduction of Aeromonas spp. was also observed.

The Effect of Mannan Oligosaccharide Supplementation on Atlantic Salmon Smolts ( Salmo salar L.) Fed Diets with High Levels of Plant Proteins

An experiment was conducted to investigate the effect of dietary mannan oligosaccharide (MOS) supplementation on Atlantic salmon ( Salmo salar L.) smolts (~ 47 g) reared in sea cages. The first treatment (control) consisted of fish fed the basal diet and the second treatment (MOS) fish were fed the same basal diet supplemented with 0.4% MOS. In the basal diet, 51.8% of the diet protein was derived from fish meal and 48.2% from vegetable protein (e.g. soy and wheat). After 14 weeks feeding on the experimental diets the results demonstrated that MOS supplementation did not affect growth performance, however, body protein composition was significantly increased. Additionally, liver histochemistry revealed that glycogen deposition in liver tissue increased from 1.80 ± 0.73 AU in the control fed fish to 2.58 ± 0.91 AU in the MOS fed fish. Histology of the anterior intestine demonstrated that MOS supplementation produced a significantly higher absorptive surface of 4.63 ± 0.62 AU compared to the control fed fish of 3.65 ±0.49 AU. The microvilli density was also significantly higher in the anterior intestine in the MOS fed fish,12.02 ± 5.95 AU, when compared to the control fed fish 5.90 ± 1.53 AU. Similar results for the absorptive area and microvilli density were observed in the posterior intestinal region.Microvilli length increased in the posterior intestine from 1.10 ± 0.18 μm in the control group to 1.41 ± 0.19 μm in the MOS fed fish. Furthermore, counts of sea lice attached to fish and total number of fish infected by sea lice were significantly lower in the MOS fed fish. The present study shows that 0.4% MOS supplementation was able to improve intestinal morphology, increase carcass protein content and glycogen deposition in the liver.

Field Observations on the Effect of a Mannan Oligosaccharide on Mortality and Intestinal Integrity of Sole (Solea senegalensis, Kaup) Infected by Photobacterium damselae subsp. piscicida

This study was conducted in order to investigate the effect of a mannan oligosaccharide (MOS) on the intestinal morphology of sole (Solea senegalensis, Kaup) reared under commercial conditions. The dietary inclusion rate for MOS was 0.4% and it was used either alone or in combination with a vaccination regime against bacterial diseases (Pasteurella spp. and Vibrio spp.). One week after the start of the experimental period, a natural outbreak of pasteurellosis, caused by Photobacterium damselae subsp.piscicida,occurred in all the groups of fish.A two-way ANOVA showed that only MOS supplementation reduced fish mortality by ca. 8% (P = 0.050). Additionally, light microscopy examination of the intestine revealed that MOS supplementation significantly increased the mucosal folding by 29% (P = 0.016) in the anterior intestinal region and by 33% (P = 0.002) in the posterior intestinalregion. Scanning electron microscopy demonstrated that both MOS supplementation and vaccination significantly increased microvilli density on the enterocyte surfaces in the anterior intestinal regionby 13% (P = 0.028) and 30% (P = 0.001) respectively. In the posterior intestinalregion neither MOS supplementation nor vaccination significantly affected the microvilli density (P = 0.005).The present study suggests that dietary MOS supplementation protects the intestinal morphology of infected sole and hinders the development of pathogenic infection, possibly by binding with Photobacterium damselae subsp. piscicida, resulting in reduced mortality of infected fish.