Insights into the fecal microbiota of captive Iberian lynx (Lynx pardinus)

The Iberian lynx (Lynx pardinus) is an endangered species restricted to several areas of Spain and Portugal. Its low genetic diversity likely provokes immune depression and high susceptibility to infectious diseases. The intestinal microbiota is closely related to host health and nutrition. In order to contribute to the knowledge of the Iberian lynx intestinal microbiota, fecal microbiota of captive specimens from two breeding centers (“La Olivilla” and “El Acebuche”), located in Southern Spain, were studied by Denaturing Gradient Gel Electrophoresis (DGGE). Results grouped microbiota in two main clusters (I and III) which included DGGE patterns of 19 out of 36 specimens, cluster I being the most frequent in “La Olivilla” (50%) and cluster III in “El Acebuche” (55.55 %) specimens. Bacteroidetes, Firmicutes and Proteobacteria phyla were identified. Segregation of clusters I and III was attributed to different microorganism presence (Pseudomonas koreensis, Pseudomonas migulae, Carnobacterium sp., Arthrobacter, Robinsoniella peorensis and Ornithinibacillus sp.) and ability to use different carbon sources. Biolog EcoPlates® results indicate high functional diversity of fecal microbiota, it being higher in cluster III. The great impact of intestinal microbiota on host health supports the importance of its microbial composition understanding. This study is the first report of captive Iberian lynx fecal microbiota composition. [Int Microbiol 20(1): 31-41 (2017)]Keywords: Iberian lynx (Lynx pardinus) · fecal microbiota · biodiversit

Detection of transposons modifying genome background in probiotics

The study of probiotic microorganisms is very interesting in the aquaculture field. Administration of live microorganisms in adequate amounts confers some benefits to the host (Kechagia et al. 2013). Even if Shewanella putrafaciens include pathogens and saprophytic strains related to fish spoilage and fish infection (Esteve, Merchán, and Alcaide 2016). The Pdp11 strain of Shewanella putrefaciens has been proved to provide beneficial effects in Sparus aurata (Chabrillón et al. 2005) and Solea senegalensis (Rodrigáñez et al. 2008). Studies focused on Pdp11 could hed light on the origin of this probiotic character. We have designed a bioinformatic workflow to detect transposons in the newly sequenced Pdp11 genome (Tapia-Paniagua et al, in press). Their presence interrupting genes account for a contribution to its probiotic character due to the lost of virulence or the gain of probiotic effect. The workflow was developed in Ruby programming language and provides: the genomic localisation of known transposons, host coding regions disrupted by complete transposons or their repeated insertion sequences, and transposons and coding regions disrupted identifiers, to stablish the putative functions of Pdp11 that could be affect by the transposons disruption. These results would support new possible hypothesis about the Pdp11 probiotic character since 14 coding regions related to S. putrefaciens were disrupted by transposons, 4 of which are directly involved in pathogenic mechanisms. This work was supported by co-funding by the European Union through the European Regional Development Fund (ERDF) 2014-2020 "Programa Operativo de Crecimiento Inteligente" together with Spanish AEI "Agencia Estatal de Investigación" to grants RTA2013-00068-C03, AGL2017-83370-C3-3-R and RTA2017-00054-C03-03.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Use of in vivo-induced antigen technology to identify bacterial genes expressed during solea senegalensis infection with Photobacterium damselae subsp. piscicida

The marine fish pathogen Photobacterium damselae subsp. piscicida (Phdp) is responsible for important outbreaks affecting several fish species including flatfish Solea senegalensis. The aim of this work was to identify in vivo-induced expressed immunogenic proteins using pooled sera from fish that have experienced photobacteriosis. In vivo induced immunogenic proteins included inosine-5'-monophosphate dehydrogenase (Impdh) and alkyl hydroperoxide reductase (AhpC), two proteins involved in peptide synthesis: serine hydroxymethyl transferase (Shmt) and alanyl-tRNA synthetase (AlaRS) and the non-ribosomal peptide synthetase involved in the synthesis of the siderophore piscibactin (Irp2)

Cambios en la microbiota intestinal de Sparus aurata tras la administración de dietas suplementadas con D. hansenii e INMUNOTEC

Abstract Effects on the intestinal microbiota of Sparus aurata after administration of diets supplemented with different doses of Debaryomyces hansenii or INMUNOTEC have been evaluated. The results revealed significant changes at metagenomical level and it could be related to metabolic changes, especially in the case of the diet supplemented with the highest dose of IMMUNOTEC. Resumen En este estudio se analizan los efectos que la inclusión en la dieta de distintas dosis de Debaryomyces hansenii e INMUNOTEC tienen sobre la microbiota intestinal de ejemplares de Sparus auratatras. Los resultados obtenidos revelaron cambios a nivel metagenómico, que podrían estar asociados con cambios metabólicos, especialmente en el caso de la dieta suplementada con la dosis más elevada de INMUNOTEC.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. Proyecto DIETAplus, JACUMAR (Junta de Cultivos Marinos, MAPAMA). Cofinanciado fondos FEM

Pathogenic strains of Shewanella putrefaciens contain plasmids that are absent in the probiotic strain Pdp11

Shewanella putrefaciens Pdp11 is a strain described as a probiotic for use in aquaculture. However, S. putrefaciens includes strains reported to be pathogenic or saprophytic to fish. Although the probiotic trait has been related to the presence of a group of genes in its genome, the existence of plasmids that could determine the probiotic or pathogenic character of this bacterium is unknown. In the present work, we searched for plasmids in several strains of S. putrefaciens that differ in their pathogenic and probiotic character. Under the different conditions tested, plasmids were only found in two of the five pathogenic strains, but not in the probiotic strain nor in the two saprophytic strains tested. Using a workflow integrating Sanger and Illumina reads, the complete consensus sequences of the plasmids were obtained. Plasmids differed in one ORF and encoded a putative replication initiator protein of the repB family, as well as proteins related to plasmid stability and a toxin-antitoxin system. Phylogenetic analysis showed some similarity to functional repB proteins of other Shewanella species. The implication of these plasmids in the probiotic or pathogenic nature of S. putrefaciens is discussed

Procedimiento de preparación, conservación, y uso en peces, del probiótico SHEWANELLA PUTREFACIENS PDP 11 para el control de enfermedades y la mejora en el crecimiento

Número de solicitud 201100385Procedimiento de preparación, conservación, y uso en peces, del probiótico Shewanella putresfaciens Pdp11 para el control de enfermedades y la mejora en el crecimiento. Preferentemente, el probiótico, compuesto por células enteras de la cepa Pdp11, se cultiva en TSAs durante 24 h a 22ºC. La preparación de una suspensión del probiótico, preferentemente sin proceso previo de liofilización o de inactivación física o química, se realiza mediante su incorporación en una matriz de alginato, preferentemente alginato sádico al 0,5%. La suspensión preparada se puede conservar sin pérdida significativa de viabilidad durante 20 - 30 días a 4ºC. Los productos alimenticios para peces son preparados mediante la adición en agitación de la suspensión de probiótico y, adicionalmente, CaCl2 50 mM, preferentemente mediante atomización.Universidad de Málaga (50%). Universidad de Almería (50%

A Tentative Study of the Effects of Heat-Inactivation of the Probiotic Strain Shewanella putrefaciens Ppd11 on Senegalese Sole (Solea senegalensis) Intestinal Microbiota and Immune Response

ABSTRACT: Concerns about safety, applicability and functionality associated with live probiotic cells have led to consideration of the use of non-viable microorganisms, known as paraprobiotics. The present study evaluated the effects of dietary administration of heat-inactivated cells of the probiotic strain Shewanella putrefaciens Ppd11 on the intestinal microbiota and immune gene transcription in Solea senegalensis. Results obtained were evaluated and compared to those described after feeding with viable Pdp11 cells. S. senegalensis specimens were fed with basal (control) diet or supplemented with live or heat inactivated (60 °C, 1 h) probiotics diets for 45 days. Growth improvement was observed in the group receiving live probiotics compared to the control group, but not after feeding with a probiotic heat-inactivated diet. Regarding immune gene transcription, no changes were observed for tnf?, il-6, lys-c1, c7, hsp70, and hsp90aa in the intestinal samples based on the diet. On the contrary, hsp90ab, gp96, cd4, cd8, il-1?, and c3 transcription were modulated after probiotic supplementation, though no differences between viable and heat-inactivated probiotic supplemented diets were observed. Modulation of intestinal microbiota showed remarkable differences based on the viability of the probiotics. Thus, higher diversity in fish fed with live probiotic cells, jointly with increased Mycoplasmataceae and Spirochaetaceae to the detriment of Brevinemataceae, was detected. However, microbiota of fish receiving heat-inactivated probiotic cells showed decreased Mycoplasmataceae and increased Brevinemataceae and Vibrio genus abundance. In short, the results obtained indicate that the viable state of Pdp11 probiotic cells affects growth performance and modulation of S. senegalensis intestinal microbiota. On the contrary, minor changes were detected in the intestinal immune response, being similar for fish receiving both, viable and inactivated probiotic cell supplemented diets, when compared to the control dietThis study was supported by MINECO (grant no. AGL2014-51839-C5-2-R and AGL2017- 83370-C3-3-R) and FEDER funds

In vitro assessment of antagonistic activities of isolates from gilthead seabream (Sparus aurata) gastrointestinal tract fed microalgae supplemented diet

The use of probiotics has emerged as a sustainable alternative to antibiotics in the control of infectious diseases, favouring fish health management, growth performance and feed utilisation, among others. microalgae represent an interesting source of nutrients and functional ingredients for aquafeeds. However, their digestibility is often limited by the presence of anti-nutritional factors or absence of appropriate enzymatic activities in the fish gastrointestinal tract (GIT). The aims of the present work were to isolate potential probiotics from the GIT of Sparus aurata fed with a diet containing 25 % microalgae and characterize their antagonism against fish pathogens. Altogether, 117 strains were isolated from juvenile seabream (146.8 ± 16.4 g) and screened for hydrolytic enzyme activities. Results showed that 48 %, 41 %, 77 % and 30 % of isolates were able to hydrolyse protein, lipids, collagen and starch, respectively. Moreover, 46 %, 8 % and 57 % of isolates exhibited the ability to degrade phytate, tannins and cellulose, respectively. Based on these results, a total of 32 isolates were selected for inhibitory activity against several fish pathogens assessment. Inhibition against Aeromonas hydrophila and Vibrio anguillarum was detected in 38 % of the isolates, whilst 44 % and 47 % inhibited P damselae subsp. damselae and P. damselae subsp. piscicida, respectively. Inhibition abilities were detected in the isolates when tested against Tenacibaculum species. Thus, 56 % inhibited Tenacibaculum maritimum; 63 % T. soleae and 22 % T. gallaecium. Overall, results showed that three strains display ability to hydrolyse 4 of the assayed substrates and produce inhibition against 8 fish pathogens, and two strains are capable to hydrolyse 5 substrates and inhibit 8 fish pathogens. Selected strains show characteristics to be considered for further characterization as potential probiotics in gilthead seabream aquaculture and microalgae-supplemented aquafeeds.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Preparado de bacterias probióticas para su administración oral a peces cultivados basado en la encapsulación en hidrogeles de alginato

Número de solicitud: 201100469La invención consiste en un preparado para la administración oral del probiótico Shewanella PDP11 a peces. El preparado se caracteriza por contener células bacterianas viables de la cepa CECT 7627 encapsuladas en un hidrogel basado en alginato cálcico en forma de partículas esféricas de morfología uniforme y tamaño modificable para adaptarse a peces de distinto tamaño. Las cápsulas contienen aditivos organolépticos y/o nutricionales que actúan como atrayentes para los animales. El preparado mantiene viables a las bacterias durante periodos prolongados, es estable en medios acuáticos, y soporta el paso por el tubo digestivo de los peces, sobre los que ejerce efectos biológicos favorables. Su administración por vía oral es independiente del alimento habitual, evitando así la inactivación del microorganismo durante la fabricación del pienso.Universidad de Almerí

Diet, Immunity, and Microbiota Interactions: An Integrative Analysis of the Intestine Transcriptional Response and Microbiota Modulation in Gilthead Seabream (Sparus aurata) Fed an Essential Oils-Based Functional Diet

Essential oils (EOs) are promising alternatives to chemotherapeutics in animal production due to their immunostimulant, antimicrobial, and antioxidant properties, without associated environmental or hazardous side effects. In the present study, the modulation of the transcriptional immune response (microarray analysis) and microbiota [16S Ribosomal RNA (rRNA) sequencing] in the intestine of the euryhaline fish gilthead seabream (Sparus aurata) fed a dietary supplementation of garlic, carvacrol, and thymol EOs was evaluated. The transcriptomic functional analysis showed the regulation of genes related to processes of proteolysis and inflammatory modulation, immunity, transport and secretion, response to cyclic compounds, symbiosis, and RNA metabolism in fish fed the EOs-supplemented diet. Particularly, the activation of leukocytes, such as acidophilic granulocytes, was suggested to be the primary actors of the innate immune response promoted by the tested functional feed additive in the gut. Fish growth performance and gut microbiota alpha diversity indices were not affected, while dietary EOs promoted alterations in bacterial abundances in terms of phylum, class, and genus. Subtle, but significant alterations in microbiota composition, such as the decrease in Bacteroidia and Clostridia classes, were suggested to participate in the modulation of the intestine transcriptional immune profile observed in fish fed the EOs diet. Moreover, regarding microbiota functionality, increased bacterial sequences associated with glutathione and lipid metabolisms, among others, detected in fish fed the EOs supported the metabolic alterations suggested to potentially affect the observed immune-related transcriptional response. The overall results indicated that the tested dietary EOs may promote intestinal local immunity through the impact of the EOs on the host-microbial co-metabolism and consequent regulation of significant biological processes, evidencing the crosstalk between gut and microbiota in the inflammatory regulation upon administration of immunostimulant feed additives.info:eu-repo/semantics/publishedVersio