Efecto de gas de formalina sobre el recuento de mesófilos en nailon comercial destinado a procedimientos quirúrgicos / Effect of formaline gas on the counting of mesophiles in commercial nylon destined to surgical procedures

RESUMEN La investigación se realizó a fin de evaluar el efecto del gas de formalina sobre el recuento de mesófilos en nailon comercial (poliamida) destinado a procedimientos quirúrgicos. En primer lugar, se evaluó la contaminación de una muestra del material comercial de 1 g expuesta al ambiente de quirófano por 72 horas, a través de la técnica de recuento en placa para mesófilos; se obtuvieron 850 UFC/g. Una vez comprobada la contaminación de la poliamida comercial, esta se sometió a gases de formalina en comprimidos. Se colocaron 5 muestras (n=5) de nailon de 1 g cada una en 5 recipientes herméticos de 1 litro con 1 gramo de formalina cada uno; estos recipientes se almacenaron en un mueble en sala de esterilización a un metro de altura del piso y posteriormente, fueron abiertos y cultivados a través de la técnica de recuento en placa para mesófilos, uno por día a lo largo de 5 días a intervalos de 24 horas. Los resultados obtenidos no registraron crecimientos de microorganismos a partir de las 24 horas y durante los 5 días posteriores y se encontraron diferencias estadísticamente significativas (p < 0,05). Se concluye que bajo las condiciones del presente estudio el tiempo de esterilización de la formalina sobre nailon comercial es de 24 horas.

ABSTRACT Hie present research work was carried out with the purpose of evaluating the effect of formalin gas on the count of mesophiles in commercial nylon (polyamide) destined to surgical procedures. Firstly, the contamination of a 1 g commercial nylon sample, exposed to the operating room environment for 72 hours, was evaluated through the plate counting technique for mesophiles; it yielded a result of 850 CFU / gram. Once the contamination of the commercial polyamide was checked, it was subjected to formalin gases in tablets. Five nylon samples (n=5) of 1 gram each were placed in 5 1 liter airtight containers containing 1 gram of formalin; Hese containers were stored in a cabinet in the sterilization room one meter above the floor, and later opened and cultivated through the plate counting technique for mesophiles, one per day for 5 days at 24 hour intervals. The results obtained after being exposed to formalin gases in tablets did not register growths of microorganisms after 24 hours and during the 5 days after the study, finding statistically significant differences (p < 0.05) in relation to the studied times concluding under the conditions of the present study that the sterilization time of the formalin on commercial nylon is equal to 24 hours.

Current Situation of Congenital Toxoplasmosis in Ecuador.

Congenital toxoplasmosis (CT) is a zoonosis resulting from the fetal transmission of the obligate intracellular parasite, T. gondii, transplacentally in pregnant women usually in the first trimester of pregnancy. This research consisted in the review of indexed articles made in the Ecuador in the period between the years 2012 and 2017 with prevalence studies of the aforementioned pathology. The purpose of this collection was to determine the current situation of this disease in the Ecuadorian territory and, in addition, to recognize the risk factors involved, affected age groups, signs and symptoms, diagnosis and prevalence of the infection. Among the risk factors were the consumption of raw or undercooked meat, contact with feces of young cats and climatic conditions that stimulate the spread of oocysts, among others. Most affected ages by this disease fluctuate between 20 and 30 years. In 90% of the cases, the disease is asymptomatic or it may present adenopathies, maculo papular erythema, hepatosplenomegaly and other general signs such as fever, malaise, headache and myalgias. According to studies conducted with specific populations, it was established that the causative agent remains latent in first-trimester pregnant women from the provinces of Pichincha (71.4% of 140 pregnant women), Guayas (73% of 5683) and El Oro (16% of 250). The results of this research evidenced missing information in Ecuador, finding few and isolated studies regarding to this pathology. Therefore, it is concluded that an updated research should be conducted in order to elucidate the true epidemiological situation of congenital toxoplasmosis in Ecuador.

Effects of dietary arachidonic acid in European sea bass (Dicentrarchus labrax) distal intestine lipid classes and gut health.

The use of low fishmeal/fish oil in marine fish diets affects dietary essential fatty acids (EFAs) composition and concentration and, subsequently, may produce a marginal deficiency of those fatty acids with a direct impact on the fish intestinal physiology. Supplementation of essential fatty acids is necessary to cover the requirements of the different EFAs, including the ones belonging to the n-6 series, such as arachidonic acid (ARA). ARA, besides its structural role in the configuration of the lipid classes of the intestine, plays an important role in the functionality of the gut-associated immune tissue (GALT). The present study aimed to test five levels of dietary ARA (ARA0.5 (0.5%), ARA1 (1%), ARA2 (2%), ARA4 (4%), and ARA6 (6%)) for European sea bass (Dicentrarchus labrax) juveniles in order to determine (a) its effect in selected distal intestine (DI) lipid classes composition and (b) how these changes affected gut bacterial translocation rates and selected GALT-related gene expression pre and post challenge. No differences were found between distal intestines of fish fed with the graded ARA levels in total neutral lipids and total polar lipids. However, DI of fish fed with the ARA6 diet presented a higher (P < 0.05) level of phosphatidylethanolamine (PE) and sphingomyelin (SM) than those DI of fish fed with the ARA0.5 diet. In general terms, fatty acid profiles of DI lipid classes mirrored those of the diet dietary. Nevertheless, selective retention of ARA could be observed in glycerophospholipids when dietary levels are low (diet ARA0.5), as reflected in the higher glycerophospholipids-ARA/dietary-ARA ratio for those animals. Increased ARA dietary supplementation was inversely correlated with eicosapentaenoic acid (EPA) content in lipid classes, when data from fish fed with the diets with the same basal composition (diets ARA1 to ARA6). ARA supplementation did not affect intestinal morphometry, goblet cell number, or fish survival, in terms of gut bacterial translocation, along the challenge test. However, after the experimental infection with Vibrio anguillarum, the relative expression of cox-2 and il-1ß were upregulated (P < 0.05) in DI of fish fed with the diets ARA0.5 and ARA2 compared with fish fed with the rest of the experimental diets. Although dietary ARA did not affect fish survival, it altered the fatty acid composition of glycerophospholipids and the expression of pro-inflammatory genes after infection when included at the lowest concentration, which could be compromising the physical and the immune functionality of the DI, denoting the importance of ARA supplementation when low FO diets are used for marine fish.

Evaluation of the analgesia nociception index for monitoring intraoperative analgesia in children.

BACKGROUND: Intraoperative analgesia is still administered without guidance. Anaesthetists decide upon dosing on the basis of mean population opioid pharmacological studies and in response to variations in haemodynamic status. However, those techniques have been shown to be imprecise. We assessed the diagnostic value of monitoring the analgesia nociception index (ANI) to detect surgical stimulation in children. METHODS: This was an observational study of 2- to 12-yr-old patients 5 min before and after surgical incision. Hypnosis was maintained with sevoflurane and guided by bispectral index. Intraoperative analgesia was administered as a remifentanil infusion titrated to variations in haemodynamic parameters, and ANI monitor values were recorded. ANI parameters assessed included instantaneous ANI (ANIi), mean ANI (ANIm), and the relative change of ANIi to ANIm (DeltaANI=ANIi-ANIm/ANIm). Statistical analyses were performed using receiver-operating-characteristic analysis with determination of the area under the receiver operating characteristic (AUROC) curve and the grey zone. RESULTS: Overall, 49 subjects were included in this study. The AUROC was 0.755 (0.738-0.772), 0.771 (0.755-0.787), and 0.756 (0.738-0.774) for ANIi, ANIm, and DeltaANI, respectively. The threshold of ANI parameters indicating the presence of noxious surgical stimuli was ≤53%, ≤56%, and ≤-13.3% for ANIi, ANIm, and DeltaANI, respectively. The percentage of subjects in the inconclusive zone was 41%, 51%, and 33% for ANIi, ANIm, and DeltaANI, respectively. CONCLUSIONS: ANI has diagnostic value for detecting surgical stimuli in children.

Feeding European sea bass (Dicentrarchus labrax) juveniles with a functional synbiotic additive (mannan oligosaccharides and Pediococcus acidilactici): An effective tool to reduce low fishmeal and fish oil gut health effects?

The aim of this study was to assess the effects of dietary mannan oligosaccharides (MOS), Pediococcus acidilactici or their conjunction as a synbiotic in low fish meal (FM) and fish oil (FO) based diets on European sea bass (Dicentrarchus labrax) disease resistance and gut health. For that purpose, sea bass juveniles were fed one of 6 diets containing different combinations of MOS (Biomos® and Actigen©; Alltech, Inc., Kentucky, USA) and Pediococcus acidilactici (BAC, Bactocell®; Lallemand Inc., Cardiff, UK) replacing standard carbohydrates as follows (MOS (%)/BAC (commercial recommendation): high prebiotic level (HP) = 0.6/0, low prebiotic level (LP) = 0.3/0, only probiotic (B) = 0/+, high prebiotic level plus probiotic (HPB) = 0.6/+, low prebiotic level plus probiotic (LPB) = 0.3/+, control (C) = 0/0 for 90 days. After 60 and 90 days of feeding trial, fish were subjected to an experimental infection against Vibrio anguillarum. Additionally, inducible nitric oxide synthase (iNOS) and tumor necrosis factor α (TNFα) gut patterns of immunopositivity and major histocompatibility complex class II (MHCII), transforming growth factor ß (TGF-ß), regulatory T-cell subset (CD4+T lymphocytes) and effector T cell (CD8α+T lymphocytes) gene expression patterns in gut by in situ hybridization were evaluated after 90 days of feeding. The effects of both additives on posterior gut through Gut Associated Lymphoid Tissue (GALT) gene expression was also studied. Fish fed the prebiotic and its combination with P. acidilactici presented increased weight regardless of the dose supplemented after 90 days of feeding, however no effect was detected on somatic indexes. For posterior gut, morphometric patterns and goblet cells density was not affected by MOS, P. acidilactici or its combination. Anti-iNOS and anti-TNFα gut immunopositivity patterns were mainly influenced by MOS supplementation and not by its combination with P. acidilactici. MHCII-ß, TCR-ß, CD4 and CD8-α positive cells distribution and incidence was not affected by diet. Fish fed HP dose presented a clear up-regulation of TNF-α, cyclooxygenase-2 (COX-2), CD4 and IL10, whereas P. acidilactici dietary supplementation increased the number of interleukin-1ß (IL1ß) and COX-2 gene transcripts. Synbiotic supplementation resulted in a reduction of MOS-induced gut humoral proinflammatory response by increasing the expression of some cellular-immune system related genes. Fish mortality after V. anguillarum infection was reduced in fish fed LPB and LP diets compared to fish fed the non-suppelmented diet after 90 days of feeding. Thus, overall pointing to the combination of a low dose of MOS and P. acidilactici as synbiont (LPB) as a viable tool to potentiate European sea bass juvenile's growth and disease resistance when supplemented in low FM and FO diets.

Supplementation of arachidonic acid rich oil in European sea bass juveniles (Dicentrarchus labrax) diets: effects on growth performance, tissue fatty acid profile and lipid metabolism.

The aim of this study was to evaluate the effects of increasing dietary arachidonic acid (ARA) levels (from 1 to 6% of total fatty acids) on European sea bass (Dicentrarchus labrax) juveniles' growth performance, tissue fatty acid profile, liver morphology as well as long-chain polyunsaturated fatty acids (LC-PUFA) biosynthesis, triglyceride and cholesterol synthesis and lipid transport. A diet with total fish oil (FO) replacement and defatted fish meal (FM) containing a 0.1-g ARA g-1 diet was added to the experimental design as a negative control diet. Dietary ARA inclusion levels below 0.2 g ARA g-1 diet significantly worsened growth even only 30 days after the start of the feeding trial, whereas dietary ARA had no effect on fish survival. Liver, muscle and whole body fatty acid profile mainly reflected dietary contents and ARA content increased accordingly with ARA dietary levels. Tissue eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA) levels were positively correlated among them. Hepatic lipid vacuolization increased with reduced dietary ARA levels. Expressions of fatty acyl desaturase 2 and 3-hydroxy-3-methylglutaryl-coenzyme genes were upregulated in fish fed the negative control diet compared to the rest of the dietary treatments denoting the influence of ARA on lipid metabolism. Results obtained highlight the need to include adequate n-6 levels and not only n-3 LC-PUFA levels in European sea bass diets.

Disease resistance and response against Vibrio anguillarum intestinal infection in European seabass (Dicentrarchus labrax) fed low fish meal and fish oil diets.

The aim of this study was to assess the effects of low levels of dietary fish meal (FM) and fish oil (FO) on disease resistance and gut associated lymphoid tissue (GALT) response after an experimental intestinal infection with V. anguillarum in European sea bass (Dicentrarchus labrax) For that purpose, sea bass juveniles were fed one of four diets containing combined levels of FO and FM as follows: 20%FM/6%FO, 20%FM/3%FO, 5%FM/6%FO and 5%FM/3%FO during 153 days. At the end of the feeding trial, fish were subjected to either an in vivo exposure to a sub-lethal dose of V. anguillarum via anal inoculation or to an ex vivo exposure to V. anguillarum. Additionally, inducible nitric oxide synthase (iNOS) and tumor necrosis factor α (TNFα) gut patterns of immunopositivity were studied. Growth performance was affected by dietary FM level, however ex vivo gut bacterial translocation rates and survival after the in vivo challenge test were affected by dietary FO level. After 5 months of feeding, low dietary FM levels led to a posterior gut up-regulation of interleukin-1ß (IL-1ß) and TNFα, major histocompatibility complex-II (MHCII) and cyclooxygenase-2 (COX2), which in turn reduced the gut associated lymphoid tissue (GALT) capacity of response after 24 h post infection and conditioned European sea bass capacity to recover gut homeostasis 7 days post infection. Immunoreactivity to anti-iNOS and anti-TNFα presented a gradient of increased immunopositivity towards the anus, regardless of the dietary FM/FO fed. Strong positive anti-TNFα isolated enterocytes were observed in the anterior gut in relation to low levels of dietary FM/FO. Submucosa and lamina propria immunoreactivity grade was related to the amount of leucocyte populations infiltrated and goblet cells presented immunopositivity to anti-iNOS but not to anti-TNFα. Thus, reducing FO content from 6% to a 3% by VO in European sea bass diets increases ex vivo and in vivo gut bacterial translocation rates, whereas reducing FM content from 20% down to 5% up-regulates the expression of several posterior gut inflammation-related genes conditioning fish growth and GALT capacity of response after bacterial infection.

Supplementation of arachidonic acid rich oil in European sea bass juveniles (Dicentrarchus labrax) diets: Effects on leucocytes and plasma fatty acid profiles, selected immune parameters and circulating prostaglandins levels.

The main objective of this study was to assess the effects of graded levels of dietary arachidonic acid (ARA), supplemented from alternative sources, on fatty acid composition of plasma and head kidney leucocytes of European sea bass (Dicentrarchus labrax). For that purpose, sea bass juveniles were fed four diets containing graded levels of ARA as follows: 0.5% (ARA0.5), 1% (ARA1), 2% (ARA2) and 4% (ARA4) during 60 days. At the end of the feeding trial fatty acid profiles of plasma and head kidney leucocytes were analyzed. Besides, plasma prostaglandins levels, head kidney leucocytes respiratory burst activity; peroxidase activity and phagocytic index were assayed. Reducing dietary ARA levels below 1% markedly reduced European sea bass growth performance. However, fish fed diet ARA0.5 tried to compensate this dietary ARA deficiency by a selective deposition of ARA on plasma and head kidney leucocytes, reaching similar levels to those fish fed diet ARA1 after 60 days of feeding. Nevertheless, head kidney phagocytic capacity was reduced as dietary ARA content in relation not only to variations on membrane composition but also to changes on fish basal prostaglandins levels. Results obtained demonstrated the importance to supply the necessary quantity n-6 LC-PUFA, and not only n-3 LC-PUFA levels, in European sea bass diets, in relation to not only growth performance but also immune system function.

Development of short-snouted seahorse (Hippocampus hippocampus, L. 1758): osteological and morphological aspects.

Information about early development after male release lags behind studies of juveniles and adult seahorses, and newborn seahorses, similar in shape to adults, are considered juveniles or fry. During early life, Hippocampus hippocampus present behavioural (shift in habitat, from planktonic to benthic) and morphological changes; for this reasons, the aims of this study are to define the stage of development of H. hippocampus after they are expelled from the male brood pouch and to establish direct or indirect development through an osteological analysis. The ossification process was studied in 120 individuals, from their release to 30 days after birth. To analyse the osteological development, Alcian Blue-Alizarin Red double staining technique for bone and cartilage was adapted to this species. At birth, H. hippocampus presents a mainly cartilaginous structure that ossifies in approximately 1 month. The bony armour composed of bony rings and plates develops in 10 days. The caudal fin, a structure absent in juveniles and adult seahorses, is present at birth and progressively disappears with age. The absence of adult osteological structure in newborns, like coronet, bony rings and plates, head spines and components allowing tail prehensile abilities, suggests a metamorphosis before the juvenile stage. During the indirect development, the metamorphic stage started inside brood pouch and followed outside and leads up to reconsider the status of H. hippocampus newborns.

Replacement of Marine Fish Oil with de novo Omega-3 Oils from Transgenic Camelina sativa in Feeds for Gilthead Sea Bream (Sparus aurata L.).

Omega-3 (n-3) long-chain polyunsaturated fatty acids (LC-PUFA) are essential components of the diet of all vertebrates. The major dietary source of n-3 LC-PUFA for humans has been fish and seafood but, paradoxically, farmed fish are also reliant on marine fisheries for fish meal and fish oil (FO), traditionally major ingredients of aquafeeds. Currently, the only sustainable alternatives to FO are vegetable oils, which are rich in C18 PUFA, but devoid of the eicosapentaenoic (EPA) and docosahexaenoic acids (DHA) abundant in FO. Two new n-3 LC-PUFA sources obtained from genetically modified (GM) Camelina sativa containing either EPA alone (ECO) or EPA and DHA (DCO) were compared to FO and wild-type camelina oil (WCO) in juvenile sea bream. Neither ECO nor DCO had any detrimental effects on fish performance, although final weight of ECO-fed fish (117 g) was slightly lower than that of FO- and DCO-fed fish (130 and 127 g, respectively). Inclusion of the GM-derived oils enhanced the n-3 LC-PUFA content in fish tissues compared to WCO, although limited biosynthesis was observed indicating accumulation of dietary fatty acids. The expression of genes involved in several lipid metabolic processes, as well as fish health and immune response, in both liver and anterior intestine were altered in fish fed the GM-derived oils. This showed a similar pattern to that observed in WCO-fed fish reflecting the hybrid fatty acid profile of the new oils. Overall the data indicated that the GM-derived oils could be suitable alternatives to dietary FO in sea bream.

Evaluation of a high-EPA oil from transgenic Camelina sativa in feeds for Atlantic salmon (Salmo salar L.): Effects on tissue fatty acid composition, histology and gene expression.

Currently, one alternative for dietary fish oil (FO) in aquafeeds is vegetable oils (VO) that are devoid of omega-3 (n-3) long-chain polyunsaturated fatty acids (LC-PUFAs). Entirely new sources of n-3 LC-PUFA such as eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids through de novo production are a potential solution to fill the gap between supply and demand of these important nutrients. Camelina sativa was metabolically engineered to produce a seed oil (ECO) with > 20% EPA and its potential to substitute for FO in Atlantic salmon feeds was tested. Fish were fed with one of the three experimental diets containing FO, wild-type camelina oil (WCO) or ECO as the sole lipid sources for 7 weeks. Inclusion of ECO did not affect any of the performance parameters studied and enhanced apparent digestibility of individual n-6 and n-3 PUFA compared to dietary WCO. High levels of EPA were maintained in brain, liver and intestine (pyloric caeca), and levels of DPA and DHA were increased in liver and intestine of fish fed ECO compared to fish fed WCO likely due to increased LC-PUFA biosynthesis based on up-regulation of the genes. Fish fed ECO showed slight lipid accumulation within hepatocytes similar to that with WCO, although not significantly different to fish fed FO. The regulation of a small number of genes could be attributed to the specific effect of ECO (311 features) with metabolism being the most affected category. The EPA oil from transgenic Camelina (ECO) could be used as a substitute for FO, however it is a hybrid oil containing both FO (EPA) and VO (18:2n-6) fatty acid signatures that resulted in similarly mixed metabolic and physiological responses.

Development of seahorse (Hippocampus reidi, Ginsburg 1933): histological and histochemical study.

Biological aspects and global demand for aquarium promote seahorses as new species with high potential for commercial purposes; however, the low newborn survival rate represents the main bottleneck of seahorses farming. In this study, the organogenesis of the Hippocampus reidi was analysed from release until the 30th day after birth, using histological and histochemical approaches. To study the stages of their early life, 360 individuals were killed, sectioned, and stained with haematoxylin and eosin, periodic acid-Schiff, and Sudan Black B techniques. At birth, mouth and anus were open, the swim bladder inflated, and the visual system highly developed. Among the results, it was emphasized the presence of the yolk sac until the 2nd day after birth, the loops of the intestine to accommodate its elongation, and the ability of the larvae to absorb lipids in the anterior and posterior tract of the intestine. A short time (7/8 days) between reabsorption of yolk sac and formation of gonads was registered, with primordial follicles visible from the 10th day after birth. For the first time, organogenesis in H. reidi was described in detail; seahorses underwent a marked metamorphosis, and the indirect development observed in this species lead up to reconsider the term "juvenile" used for H. reidi during this period.

Preliminary insights into the incorporation of rosemary extract (Rosmarinus officinalis L.) in fish feed: influence on performance and physiology of gilthead seabream (Sparus aurata).

Gilthead seabream (Sparus aurata) were fed a basal (control) diet and four experimental diets (R600, R1200, R1800 and R2400), containing 600, 1200, 1800 and 2400 mg kg(-1), respectively, of rosemary extract (Rosmarinus officinalis L.). At 4 and 12 weeks from the beginning of the ongrowing period, the fish were sacrificed, blood was drawn to obtain plasma and the liver and intestines were dissected. Growth and feed intake were unaffected by rosemary extract addition. A histological examination of the intestine revealed no differences among the dosages, while the liver showed a sharp decrease in hepatic steatosis in diets supplemented with rosemary extract. Furthermore, plasma alanine aminotransferase was lower with these diets at the end of the ongrowing period. Rosemary extract reduced the plasma levels of glucose and triglycerides on week 4 and glucose and HDL/LDL cholesterol ratio on week 12, suggesting better transport and energy metabolism of the lipids. Overall, the most evident effect of rosemary extract was observed with the 600 mg kg(-1) dose.

Streptococcus iniae in gilthead seabream (Sparus aurata, L.) and red porgy (Pagrus pagrus, L.): ultrastructural analysis.

Streptococcosis caused by Streptococcus iniae has become one of the most serious marine and freshwater aquatic diseases in the past decade, causing large losses in farmed and wild fish worldwide. In this study, we performed an ultrastructural study of major lesions in gilthead seabream Sparus aurata and red porgy Pagrus pagrus experimentally infected with the S. iniae IUSA-1 strain, isolated in a natural outbreak in Spain in the mentioned species. The transmission electron micrographs revealed the resistance of this pathogen inside the phagosome, indicating that the macrophage may provide a significant bacterial reservoir for continuing infection, disease dissemination, and tissue injury by crossing the blood-brain barrier.

Effects of dietary DHA and α-tocopherol on bone development, early mineralisation and oxidative stress in Sparus aurata (Linnaeus, 1758) larvae.

DHA deficiency has been related to skeletal malformations in fish, but high DHA levels have produced controversial results that could relate to the oxidative status of fish tissues in the different reports. In the present study, gilthead seabream (Sparus aurata) larvae were fed deficient, adequate or high DHA levels, or high DHA levels supplemented with the antioxidant α-tocopherol. Larvae fed deficient DHA levels tended to be smaller, and showed the highest incidence of urinary bladder calculi, lordosis and kyphosis and the lowest number of mineralised vertebrae for any given size class. Elevation of dietary DHA increased larval growth and significantly enhanced the expression of the insulin-like growth factor 1 (IGF-1) gene. However, a DHA level increase up to 5 % raised the degree of lipid oxidation in larval tissues and deformities in cranial endochondral bones and in axial skeletal haemal and neural arches. The increase in dietary α-tocopherol supplementation in high-DHA feeds reduced again the occurrence of skeletal deformities. Moreover, the expression of genes coding for specific antioxidants such as catalase, superoxide dismutase or glutathione peroxidase, which neutralised reactive oxygen substances formed by increased dietary DHA, was significantly decreased in larvae fed high α-tocopherol levels. These results denoted the importance of DHA for early bone formation and mineralisation. Low dietary DHA levels delay early mineralisation and increase the risk of cranial and axial skeletal deformities. Excessive DHA levels, without an adequate balance of antioxidant nutrients, increase the production of free radicals damaging cartilaginous structures before bone formation.

Oxidative status and histological changes in sea bass larvae muscle in response to high dietary content of docosahexaenoic acid DHA.

In previous studies, we observed dystrophic alterations in muscle of 48-day-old sea bass fed imbalanced docosahexaenoic acid (DHA) and vitamin E diets. To understand the whole pathological process associated with oxidative stress, a histological study was performed by feeding 14-day-old sea bass larvae with microdiets containing different ratios of DHA/vitamin E (1/150, 5/150 and 5/300) for a period of 21 days. Larvae fed diet 1/150 showed no lesions in contrast to larvae fed diets 5/150 and 5/300 where the highest incidence of muscle lesions and thiobarbituric acid reactive substances (TBARS) content was observed. Semithin sections revealed focal lesions consisting of degenerated fibres with hypercontracted myofilaments and extensive sarcoplasm vacuolization affecting both red and white muscle. Ultrathin sections of degenerating muscle fibres showed diffuse dilatation of sarcoplasmic reticulum, disorganized myofilaments and autophagic vacuoles containing myelin figures and dense bodies. Additionally, some macrophages were observed among injured fibres as numerous satellite cells. Results from the study agree with those obtained from previous work, proving the pathological potential of free radicals in sea bass larvae musculature. Moreover, high vitamin E inclusion could not completely protect cell membranes from free radicals action.

Effects on mortality and stress response in European sea bass, Dicentrarchus labrax (L.), fed mannan oligosaccharides (MOS) after Vibrio anguillarum exposure.

The effects of dietary mannan oligosaccharides (MOS; 4 g kg(-1) ; Bio-Mos, Alltech Inc, USA) in diets for European sea bass, Dicentrarchus labrax (L.), juveniles in relation to disease and stress resistance, combining intestinal infection with Vibrio anguillarum and stress challenge by confinement, were assessed in this study. After 8 weeks of MOS supplementation, fish were exposed to a pathogen challenge test against V. anguillarum by direct gut inoculation combined with a confinement stressor panel. Cumulative mortality of fish fed MOS caused by anally inoculated V. anguillarum decreased from 66% to 12.5% and from 54.1% to 25% in infected and infected + stressed fish, respectively, compared to fish fed control diet. Results for European sea bass revealed a positive effect of MOS dietary inclusion on disease resistance, in terms of cumulative mortality, against gut inoculated V. anguillarum, as well as reduced effects of stress on microbiota diversity. Both of these findings, together with the enhanced innate immune response and the higher gut mucus production and density of eosinophil granulocytes in gut mucosa obtained in previous studies after MOS supplementation (Torrecillas et al. 2007, 2011a,b) suggest that general reinforcement of the innate immune system, and particularly of the intestinal barrier efficiency, is the main defence mechanism of European sea bass fed MOS against pathogenic microorganisms.