Effects of a dietary ß-(1,3)(1,6)-D-glucan supplementation on intestinal microbial communities and intestinal ultrastructure of mirror carp (Cyprinus carpio L.).

AIM: To assess the effects of dietary Saccharomyces cerevisiae ß-(1,3)(1,6)-D-glucan supplementation (MacroGard(®)) on mirror carp (Cyprinus carpio L.) intestinal microbiota and ultrastructure of the enterocyte apical brush border. METHODS AND RESULTS: Carp were fed either a control diet or diets supplemented with 0.1, 1 or 2% w/w MacroGard(®). Culture-dependent microbiology revealed that aerobic heterotrophic bacterial levels were unaffected by dietary MacroGard(®) after 2 and 4 weeks. No effects were observed on the allochthonous lactic acid bacteria (LAB) populations at either time point; however, reduced autochthonous LAB populations were observed at week 4. PCR-DGGE confirmed these findings through a reduction in the abundance of autochthonous Lactococcus sp. and Vagococcus sp. in MacroGard(®)--fed fish compared with the control-fed fish. Overall, sequence analysis detected microbiota belonging to the phyla Proteobacteria, Firmicutes, Fusobacteria and unidentified uncultured bacteria. DGGE analyses also revealed that dietary MacroGard(®) reduced the number of observed taxonomical units (OTUs) and the species richness of the allochthonous microbiota after 2 weeks, but not after 4 weeks. In contrast, dietary MacroGard(®) reduced the number of OTUs, the species richness and diversity of the autochthonous microbiota after 2 weeks, and those parameters remained reduced after 4 weeks. Transmission electron microscopy revealed that intestinal microvilli length and density were significantly increased after 4 weeks in fish fed diets supplemented with 1% MacroGard(®). CONCLUSIONS: This study indicates that dietary MacroGard(®) supplementation modulates intestinal microbial communities of mirror carp and influences the morphology of the apical brush border. SIGNIFICANCE AND IMPACT OF THE STUDY: To the authors' knowledge, this is the first study to investigate the effects of ß-(1,3)(1,6)-D-glucans on fish gut microbial communities, using culture-independent methods, and the ultrastructure of the apical brush border of the enterocytes in fish. This prebiotic-type effect may help to explain the mechanisms in which ß-glucans provide benefits when fed to fish.

The effect of Pediococcus acidilactici on the gut microbiota and immune status of on-growing red tilapia (Oreochromis niloticus).

AIM: To assess Pediococcus acidilactici as a dietary supplement for on-growing red tilapia (Oreochromis niloticus). METHODS AND RESULTS: Tilapia were fed either a control diet or control diet supplemented with Ped. acidilactici at 10(7) CFU g(-1) for 32 days. Ped. acidilactici colonized the intestinal tract and significantly affected the intestinal microbial communities. PCR-DGGE revealed direct antagonism of gastric Ped. acidilactici with an endogenous uncultured bacterium during a period of reverting to nonsupplemented feeding. Light microscopy revealed that gut integrity and leucocyte levels were unaffected by Ped. acidilactici; however, blood leucocyte levels and serum lysozyme activity were elevated after 14-days' feeding. No significant improvements in growth performance were observed at the end of the trial (day 32), but survival was significantly higher in the probiotic group. CONCLUSIONS: The study demonstrates that oral supplementation of Ped. acidilactici modulates intestinal bacterial communities in on-growing red tilapia and also stimulates some aspects of the nonspecific immune response. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge this is the first study assessing the effects of probiotics on the gut microbiota of tilapia using culture-independent methods. Such methods are crucial to understand the mechanisms which underpin and mediate host benefits.

The morphology of the inner ear from the domestic pig (Sus scrofa).

The morphology of the hair cells of the inner ear end organs from the domestic pig (Sus scrofa) have been studied using a combination of Scanning and Transmission Electron Microscopy (SEM and TEM), revealing hair cells from the cochlea and vestibule using a novel surgical and technical approach. This is the first time that the inner ear hair cells from S. scrofa have been studied, thus providing useful anatomical information on the morphology of the hair cells from the cochlea, saccule and utricle from a large mammal. Anatomical information in relation to the morphology of the inner ear is of considerable importance, both in the pathological diagnosis of trauma and in the development of cochlea implants and other biotechnological systems associated with the enhancement of hearing. Standard fixation protocols using cardiac perfusion was not employed in this study as this method cannot always be applied, such as the pathological examination of the human ear, or the study of animals protected by endangered species legislation. With the exception of a very few countries, cetaceans cannot be killed for research purposes, yet physiological information on the inner ear from these species is urgently required for ecological assessment reasons. Supporting the use of S. scrofa as a model for cetacean hearing research is that this animal is a member of the order Artiodactyla, which includes both the hippopotamus and cetaceans. Being of a similar size, the pig is an ideal subject for developing protocols and surgical techniques required to investigate both the human and small cetacean auditory systems.

The inner ear ultrastructure from the paddlefish (Polyodon spathula) using transmission electron microscopy.

The structure of the hair cells on each sensory macula from the inner ear of the paddlefish (Polyodon spathula) was studied using scanning and transmission electron microscopy, revealing the nucleated cell bodies and peripheral nerve fibres of the saccule utricle and lagena. Examination of the structures within the cell body revealed comparable features with those found in the inner ear hair cells from bony fish species, although in P. spathula the afferent cell body is almost twice the size. This is the first time that the inner ear hair cells from an Acipenseriform fish have been studied using transmission microscopy, thus providing benchmark anatomical information in relation to the cellular morphology of the afferent receptors from a 'healthy' P. spathula ear. Structural information is of assistance in the study of aquatic animal hearing for environmental monitoring purposes, as morphological data can be used to confirm if evidence of raised hearing thresholds from animals exposed to intense anthropogenic noise or other destructive agents (determined using electrophysiological or behavioural techniques) are a direct result of damage to the ultrastructure of the inner ear.