Effect of dietary inclusion of microalgae (Nannochloropsis gaditana and Schizochytrium spp) on non-specific immunity and erythrocyte maturity in Atlantic salmon fingerlings.

The parr-smolt transformation in salmonids involves a critical period characterized by systemic changes associated with the fish's immune response. In this context, as a dietary ingredient in functional diets, microalgae offer an alternative due to their nutritional and bioactive compounds that could strengthen the immune status. This study evaluated the effect of a diet supplemented with Schizochytrium spp and Nannochloropsis gaditana on the expression of genes associated with the antibacterial response. Additionally, the study assessed the effect on the leukocyte population and erythrocyte maturity in Salmo salar blood. Fish were fed for 30 days with a microalgal mixture (1:1) at a 10% inclusion. Each diet was randomly assigned to a tank using a completely randomized design (CRD) with four replications. Each tank was stocked with 70 Atlantic salmon fingerlings with an initial mean weight of 78.87 ± 0.84. Transcription levels were quantified and analyzed by qRT-PCR from cell isolates and mucus tissue. Furthermore, cell count and identification of leukocytes and classification of cellular maturity of erythrocytes using a neural network with a multilayer perceptron (MLP) were performed. Our results showed a significant (p < 0.05) increase in fold change expression of C3 (2.54 ± 0.65) and NK-Lysine (6.84 ± 0.94) in erythrocytes of microalgae-supplemented fish. Moreover, a significant increase of 1.59 and 2.35 times in monocytes and immature erythrocytes, respectively, was observed in the same group of fish (p < 0.05). This study's results indicate that dual microalgae (Schizochytrium spp and N. gaditana) supplementation can increase innate humoral antibacterial components, particularly in erythrocyte tissue, and increase phagocytic cells and immature erythrocytes in S. salar blood.

Herbivorous fish (Medialuna ancietae) as a sustainable alternative for nutrition security in Northern Chile.

Sustainability in aquaculture is a necessity of the future, not only as the most promising means of supplying the protein that the world will require to feed its growing population but to offer needed conservation of the world's ocean resources. The use of wild fish inputs in farm-raised fish outputs has been a primary concern of sustainability in aquaculture production. Herbivorous fish are more efficient converters of protein into fish flesh. Species of the genus Medialuna fish have been reported as a fast-growing, short-lived species. The native fish Acha (Medialuna ancietae Chirichigno 1987) in the Northern part of Chile is an over-exploited fish that has been associated with aquatic vegetation as a food source. We studied the feeding habits and nutritional composition of M. ancietae. For this, we developed a reference collection of marine macroalga (epidermis and nutritional composition) observed in the diet of individuals of this species for the study of digestive material. More than 90% of the components found were marine macroalgae, indicating that M. ancietae is an herbivorous fish. Compared to non-herbivorous fish our results showed that most of the nutrients present in the Medialuna diet are found at much lower levels including n-3 long-chain polyunsaturated fatty acids (49.7%) and protein (13-60%). M. ancietae meat provides essential components of human nutrition with a significant protein content (18.99 ± 0.26%) and 268 ± 5.9 mg/100 g of the essential n-3 long-chain polyunsaturated fatty acids. Most fed aquaculture non-herbivorous species rely on wild-captured fish for these essential nutrients, while M. ancietae can obtain and concentrate them from potentially cultivable macroalgae. M. ancietae has potential for sustainable aquaculture production as a contribution to nutrition security and the re-stocking of wild populations.

House fly (Musca domestica) larvae meal as an ingredient with high nutritional value: Microencapsulation and improvement of organoleptic characteristics.

Insects have potential to become food ingredients, but it is necessary to improve the sensory properties of insects to help them to be better accepted by the population. The purpose of this study was to produce and characterize house fly larval meal (FLM) converted to a micro-encapsulated powder to improve appearance and other organoleptic characteristics. FLM showed high protein (54%) and lipid (22%) content, with a microbiological activity compatible for food purposes. Moreover, the high content of essentials amino acids (lysine, cysteine and leucine) and unsaturated fatty acids (oleic, linoleic and palmitoleic) make FLM a valuable nutritional source. Spray drying was selected to encapsulate FLM (0.5-2% w/v) using maltodextrin (20% w/v) and alginate (0.5% w/v). Encapsulation improved the appearance of FLM, creating a white-beige, monodispersed micro-powder (9 µm in size). Micro-powder with 2% FLM is considered a good source of protein (5.1%). Microencapsulation also dramatically reduced the volatile emissions of FLM. In conclusion, novel FLM micro-powders were developed using a simple and scalable encapsulation technique. The micro-powder with 2% FLM is a good source of protein, has a pleasant appearance similar to vegetable meals and has improved odor compared to typical insect meals. Thus, insect-based food ingredients in micro-powders could become more accepted by the general population.

Increased dietary availability of selenium in rainbow trout (Oncorhynchus mykiss) improves its plasma antioxidant capacity and resistance to infection with Piscirickettsia salmonis.

Salmonid Rickettsial Septicaemia (SRS), caused by Piscirickettsia salmonis, is the most important infectious disease in the Chilean salmon farming industry. An opportunity to control this disease is to use functional micronutrients to modulate host mechanisms of response to the infection. Since P. salmonis may affect the host antioxidant system in salmonids, particularly that dependent on selenium (Se), we hypothesized that fish's dietary selenium supplementation could improve the response to the bacterial infection. To address this, we defined a non-antibiotic, non-cytotoxic concentration of selenium to evaluate its effect on the response to in vitro infections of SHK-1 cells with P. salmonis. The results indicated that selenium supplementation reduced the cytopathic effect, intracellular bacterial load, and cellular mortality of SHK-1 by increasing the abundance and activity of host glutathione peroxidase. We then prepared diets supplemented with selenium up to 1, 5, and 10 mg/kg to feed juvenile trout for 8 weeks. At the end of this feeding period, we obtained their blood plasma and evaluated its ability to protect SHK-1 cells from infection with P. salmonis in ex vivo assays. These results recapitulated the observed ability of selenium to protect against infection with P. salmonis by increasing the concentration of selenium and the antioxidant capacity in fish's plasma. To the best of our knowledge, this is the first report of the protective capacity of selenium against P. salmonis infection in salmonids, becoming a potential effective host-directed dietary therapy for SRS and other infectious diseases in animals at a non-antibiotic concentration.

Antimicrobial resistance in Chilean marine-farmed salmon: Improving food safety through One Health.

Aquaculture is seen as an essential requirement for improving food security and nutrition. Fish such as salmonids are a primary source of protein and essential nutrients. Aquaculture provide income for communities across the world and have a smaller carbon footprint than terrestrial animal-production systems. However, fish diseases are a constant threat, and the use of antibiotics is a source of concern due to its adverse impacts on the environment and human health. Chilean salmon farming has made several efforts to reduce the use of antibiotics for the eradication of piscirickettsiosis, a disease caused by the gram-negative bacteria Piscirickettsia salmonis. Excessive amounts of antibiotics continue to be used in Chilean aquaculture, playing an important role in the emerging public health crisis of antimicrobial resistance. Without doubt, P. salmonis is becoming increasingly resistant to important frontline antimicrobial classes, with severe implications for the future treatment of infectious human and animal diseases. Antimicrobial-resistant bacteria as well as antibiotic residues from salmon production are spreading in the environment, and thus both salmon food commodities and wild organisms can become a source of resistant bacteria that can be transmitted to humans as foodborne contaminants. This urgent threat needs to be addressed by implementing national strategies in compliance with international standards that include both prudent antimicrobial use in marine salmon farms and the investment towards a One Health approach, which combines human, animal and environmental health.

Pharmacological iron-chelation as an assisted nutritional immunity strategy against Piscirickettsia salmonis infection.

Salmonid Rickettsial Septicaemia (SRS), caused by Piscirickettsia salmonis, is a severe bacterial disease in the Chilean salmon farming industry. Vaccines and antibiotics are the current strategies to fight SRS; however, the high frequency of new epizootic events confirms the need to develop new strategies to combat this disease. An innovative opportunity is perturbing the host pathways used by the microorganisms to replicate inside host cells through host-directed antimicrobial drugs (HDAD). Iron is a critical nutrient for P. salmonis infection; hence, the use of iron-chelators becomes an excellent alternative to be used as HDAD. The aim of this work was to use the iron chelator Deferiprone (DFP) as HDAD to treat SRS. Here, we describe the protective effect of the iron chelator DFP over P. salmonis infections at non-antibiotic concentrations, in bacterial challenges both in vitro and in vivo. At the cellular level, our results indicate that DFP reduced the intracellular iron content by 33.1% and P. salmonis relative load during bacterial infections by 78%. These findings were recapitulated in fish, where DFP reduced the mortality of rainbow trout challenged with P. salmonis in 34.9% compared to the non-treated group. This is the first report of the protective capacity of an iron chelator against infection in fish, becoming a potential effective host-directed therapy for SRS and other animals against ferrophilic pathogens.

Encapsulación de extractos antioxidantes desde sub-productos agroindustriales: una revisión / Encapsulation of antioxidant extracts from agroindustrial by-products: a review

RESUMEN Actualmente se generan grandes cantidades de sub-productos agroindustriales como residuos, aun cuando pueden ser fuente importante de variados compuestos antioxidantes. Una de las formas de aprovechar esta funcionalidad es concentrar los antioxidantes mediante la elaboración de extractos, siendo las "extracciones verdes", que usan solventes como agua o etanol, que por ser más amigables con el medio ambiente y la salud humana, han sido las más estudiadas en los últimos años. Sin embargo, uno de los principales problemas de reutilizar este tipo de compuestos bioactivos es mantener su estabilidad, debido a que luego de ser extraídos desde su matriz biológica son muy sensibles a distintas condiciones medioambientales y de almacenamiento. Una tecnología que puede reducir la inestabilidad de los compuestos antioxidantes es la encapsulación, la cual también reduce las alteraciones organolépticas que se pudieran producir cuando son incorporados en alimentos. Según nuestro conocimiento no existen revisiones de encapsulación de extractos de sub-productos agroindustriales (en donde se puede encontrar una gran variedad de compuestos antioxidantes) y la información existente se basa en compuestos antioxidantes específicos encapsulados. Por tanto, el objetivo de esta revisión fue recopilar información actualizada respecto a la encapsulación de extractos antioxidantes obtenidos a partir de sub-productos agroindustriales vegetales y su incorporación como ingredientes en alimentos.

ABSTRACT Currently large quantities of agro-industrial by-products are generated as waste even when they can be used as important sources of various antioxidant compounds. One of the ways to take advantage of this functionality is to concentrate antioxidants via the preparation of extracts. Due to their lower impact on the environment and human health "green extractions" using solvents such as water or alcohol have been the most studied in recent years. However, one of the main problems in re-using these bioactive compounds is maintaining stability after being extracted from their biological matrix. Instability relates to different environmental and storage conditions. One technology that can reduce antioxidants instability is encapsulation, which also reduces organoleptic alterations that could occur when they are incorporated into food. To our knowledge there are no reviews of encapsulation of agroindustry by-product extracts (where a wide variety of antioxidant compounds can be found) and existing information is based on specific encapsulated antioxidant compounds. Therefore, the objective of this review was to gather up-to-date information regarding the encapsulation of antioxidant extracts obtained from vegetable agro-industrial by-products and their incorporation as ingredients in food.

Fermented Soybean Meal Increases Lactic Acid Bacteria in Gut Microbiota of Atlantic Salmon (Salmo salar).

The main goal of the present study was to address the effect of feeding fermented soybean meal-based diet to Atlantic salmon on gut microbiota. Further, expression of genes of interest, including cathelicidin antimicrobial peptide (cath), mucin 2 (muc2), aquaporin (aqp8ab), and proliferating cell nuclear antigen (pcna), in proximal intestine of fish fed either experimental diet was analyzed. Three experimental diets, including a control fishmeal (30% FM), soybean meal (30% SBM), or fermented soybean meal diet (30% FSBM) were randomly assigned to triplicate tanks during a 50-day trial. The PCR-TTGE showed microbiota composition was influenced by experimental diets. Bands corresponding to genus Lactobacillus and Pediococcus were characteristic in fish fed the FSBM-based diet. On the other hand, bands corresponding to Isoptericola, Cellulomonas, and Clostridium sensu stricto were only observed in fish FM-based diet, while Acinetobacter and Altererythrobacter were detected in fish fed SBM-based diet. The expression of muc2 and aqp8ab were significantly greater in fish fed the FSBM-based diet compared with the control group. Our results suggest feeding FSBM to Atlantic salmon may (1) boost health and growth physiology in fish by promoting intestinal lactic acid bacteria growth, having a prebiotic-like effect, (2) promote proximal intestine health by increasing mucin production, and (3) boost intestinal trans-cellular uptake of water. Further research to better understands the effects of bioactive compounds derived from the fermentation process of plant feedstuff on gut microbiota and the effects on health and growth in fish is required.

Effect of anthocyanidins on myogenic differentiation in induced and non-induced primary myoblasts from rainbow trout (Oncorhynchus mykiss).

A study was conducted to test whether an anthocyanidin mixture (peonidin, cyanidin and pelargonidin chloride) modulates myogenesis in both induced and non-induced myogenic cells from juvenile rainbow trout (Oncorhynchus mykiss). We evaluated three different anthocyanidin concentrations (1×, 2.5× and 10×) at two sampling times (24 and 36h). To test for treatment effects, we analyzed the expression of myoD and pax7 as well as two target genes of the Notch signaling pathway, hey2 and her6. In induced myogenic cells, the lowest and middle anthocyanidin doses caused significantly greater expression of myoD after 24h of treatment compared to control. A significantly higher expression of pax7 in cells exposed to either anthocyanidin treatment during 36h compared was observed. Similarly, the pax7/myoD ratio was significantly lower in cells exposed to the lowest anthocyanidin doses during 24h compared to control. No significant effect of anthocyanidin treatments on the expression of hey2 and her6 at either sampling point was detected. In non-induced cells, we observed no effect of anthocyanidins on myoD expression and significant down-regulation on pax7 expression in cells exposed to either anthocyanidin mixture concentrations after 24 and 36h of treatment compared to control. Further, the pax7/myoD ratio was significantly lower in cells exposed to either anthocyanidin doses at both sampling time. In non-induced cells, the highest anthocyanidin dose provoked significantly greater expression of hey2 after 24h of treatment compared to control. We detected no such effect in non-induced cells exposed to the lowest and middle anthocyanidin doses during 24h of treatment. The expression of her6 was unaffected by anthocyanidin treatments at either sampling time or doses compared to control. Collectively, these findings provide evidence that anthocyanidins modulate specific components of the myogenic programming in fish, thereby potentially affecting somatic growth in fish fed plant-derived extracts rich in this type of polyphenols. Moreover, in early differentiating myogenic cells, the anthocyanidin effect on myogenic programming appears to differ based upon the exposure time and the differentiation stage of the myogenic cells by boosting myogenic differentiation signaling after 24h treatment while pausing differentiation, potentially favoring cell survival after 36h treatment. Further research to determine whether plant-derived secondary metabolites including alkaloids, terpenoids, tannins, saponins, glycosides, flavonoids, phenolics, steroids and essential oils can modulate myogenic programming in myogenic cells isolated from finfish species is warranted.

Immunomodulatory effect of cathelicidins in response to a ß-glucan in intestinal epithelial cells from rainbow trout.

The aim of the present study was to characterize intestinal immune mechanisms involved in the response to ß-glucans in rainbow trout. Among the immune effectors regulated in response to immunostimulants, host defense peptides (HDPs) are abundantly expressed in epithelial linings, suggesting their important role in the mucosal immune response. Therefore, the immunomodulatory properties of expressed HDPs in the epithelial intestinal cells of rainbow trout in response to the ß-glucan, zymosan, were assessed. The results showed that zymosan increased the production of the HDP, cathelicidin, and the cytokine, IL-1ß, in the intestinal epithelial RTgutGC cell line at the transcript and protein levels. Thus, cathelicidin-2 variants were produced and were shown to (i) induce the production of IL-1ß in RTgutGC cells and (ii) display a synergic effect with zymosan in IL-1ß upregulation. Importantly, the colocalization of both rtCATH-2 and IL-1ß was detected in the intestinal epithelial cells of rainbow trout fed with a 0.3% zymosan-supplemented diet. We propose that trout cathelicidins are expressed by intestinal epithelial cells and exert immunomodulatory effects to improve the local intestinal immune response triggered by immunostimulants.

Caracterización histológica y morfológica del tracto digestivo de la merluza austral (Merluccius australis) / Histological and morphological characterization of the digestive tract of southern hake (Merluccius australis)

Las características anatómicas e histológicas del tracto alimentario de la merluza austral (Merluccius australis) se han estudiado con el fin de comprender la fisiología digestiva y formular alimentos balanceados adecuados. M. australis posee un esófago corto, un estómago en forma de y grande y un intestino corto. Histológicamente, el tracto digestivo está formado por cuatro capas: la mucosa, submucosa, muscularis y la serosa. El esófago es un órgano tubular corto de pared gruesa con una gran capacidad de distensión y abundantes células caliciformes secretoras de mucus que permite el paso de grandes presas. El estómago es grande y musculoso, con alta capacidad de distensión. No se observo la presencia de apéndices pilóricos. El intestino es corto y dividido en regiones: proximal, medio y distal. En la región anterior del intestino se observaron abundantes células caliciformes, que disminuyeron en la región media para finalmente dispersarse en la región posterior. Además, los enterocitos mostraron un aumento en la presencia de vacuolas supranuclear en la región posterior del intestino. Los resultados de este estudio anatómico e histológico indican que M. australis tiene un sistema digestivo similar a otros peces gadiformes, el cual está adaptado para digerir y absorber presas ricas en proteínas, principalmente pescados y crustáceos.

The anatomy and histology features of alimentary tract of southern hake (Merluccius australis) have been studied in order to understand the digestive physiology and formulate suitable feeds. M. australis has a short oesophagus, large y-shaped stomach and short intestine, and histologically, the digestive tract is formed by four layers: mucosa, submucosa, muscularis and serosa. The oesophagus is a short tubular thick-walled organ with a great distension capacity and abundant mucus secreting goblet cells allowing the passage of large preys. The stomach is large and muscular with high capability for distension. Pyloric caeca were not present. The intestine is short and divided into a proximal middle and distal segment. In the anterior region of the intestine was observed abundant goblet cells, which decreased in the middle region and finally dispersed in the posterior region. Additionally, enterocytes showed an increase in the presence of supranuclear vacuoles at the posterior region of the intestine. The results of this anatomical and histological study indicate that M. australis has a digestive tract similar to other gadiform fishes, which is adapted to digest and absorb protein-rich preys, mainly fish and crustacean.

Effect of starvation on transcriptomes of brain and liver in adult female zebrafish (Danio rerio).

We used microarray and quantitative real-time PCR (qRT-PCR) analyses in adult female zebrafish (Danio rerio) to identify metabolic pathways regulated by starvation in the liver and brain. The transcriptome of whole zebrafish brain showed little response to 21 days of starvation. Only agouti-related protein 1 (agrp1) significantly responded, with increased expression in brains of starved fish. In contrast, a 21-day period of starvation significantly downregulated 466 and upregulated 108 transcripts in the liver, indicating an overall decrease in metabolic activity, reduced lipid metabolism, protein biosynthesis, proteolysis, and cellular respiration, and increased gluconeogenesis. Starvation also regulated expression of many components of the unfolded protein response, the first such report in a species other than yeast (Saccharomyces cerevisiae) and mice (Mus musculus). The response of the zebrafish hepatic transcriptome to starvation was strikingly similar to that of rainbow trout (Oncorhynchus mykiss) and less similar to mouse, while the response of common carp (Cyprinus carpio) differed considerably from the other three species.

Effect of cholinergic agonists on muscular tonus of the lizard small intestine and esophagus

The underlying mechanisms of acetycholine-induced intestinal relaxation in the lizard Liolaemus tenuis tenuis are still unknows. By using a classical model of intestinal recording of isometric contraction and relaxation in conjunction with specific pharmacological tools, this article studies the possible influence of EDRF/NO and nicotinic ganglionar receptors on the Ach-induced relaxation in an effort to elucidate the probable mechanisms involved in ACh effect. It was observed that the relaxation of the lizard intestine elicited by ACh (10(-7) - 4 x 10(-4) M) was not affected by hexametonium (5 x 10(4) M) or tetrodotoxin (10(-6) M). Nicotine (10(-7) to 10(-4) M) induced relaxation was significantly antagonized by hexametonium; however, it was not influenced by tetrodotoxin. These results allow us to discard a neuronal pathway in cholinergic-induced relaxation, suggesting a more direct cholinergic effect on the smooth muscle, perhaps mediated by an unknown substance released by some specialized tissue. N-nitro-L-arginine, used to block NO-synthase and NO production, induced no changes in ACh-induced relaxation. Methylene blue, a soluble guanylate cyclase inhibitor, induced no changes in ACh-induced relaxation. These results allow us to dicard a probable role of EDRF/nitric oxide in the ACh-induced relaxation of lizard small intestine, providing evidence that this mechanism could be different from reported on other species.

Relación entre desplazamiento de melanosomas y cambios de color en lagartijas machos: liolaemus tenuis tenuis: un estudio ultraestructural / Relationship between melanosomes displacement and color change in male lizards: liolaemus tenuis tenuis: an ultrastructural study

La mayor parte de los organismos multicelulares se caracterizan por un patrón de color distintivo y, a menudo, particular. Uno de los efectores relacionados con el proceso corresponde a células especiales, los cromatóforos. En este trabajo se demuestra mediante técnicas de microscopía electrónica, que en el dermis de reptiles sometidos a 35§C por 12 min, se produce una fuerte concentración de melanosomas en torno al núcleo de los melanóforos. Por el contrario, a 0§ los melanóforos presentan expansiones citoplasmáticas que contienen numerosos melanosomas. Asociados a los melanosomas en dispersión, se observan microtúbulos. Esta característica permite plantear la posible existencia de algún tipo de relación entre microtúbulos y movimiento de melanosomas en este reptil, cuando es sometido a bajas temperaturas, momento en que se oscurecen. Se destacan también, gruesos manojos tridimensionales de fibras colágenas asociadas a fibroblastos, estructuras, que para algunos autores, representarían un primordio de esclerificación dérmica, fenómeno que predecería a la fase fibrilar de la osificación dérmica o directa. Otro tipo de célula, los iridóforos, contienen placas reflectantes que reflejan la luz en direcciones determinadas