Characterization of the impact of dietary immunostimulant CpG on the expression of mRNA biomarkers involved in the immune responses in Atlantic salmon (Salmo salar).

Infectious diseases have significantly impacted Atlantic salmon aquaculture worldwide. Modulating fish immunity with immunostimulant-containing functional feeds could be an effective strategy in mitigating disease problems. Previously, we characterized the impact of polyriboinosinic polyribocytidylic acid (pIC) and formalin-killed typical Aeromonas salmonicida bacterin on miRNA expression in Atlantic salmon fed a commercial diet with and without immunostimulant CpG. A set of miRNA biomarkers of Atlantic salmon head kidney responding to pIC and/or bacterin immune stimulations was identified (Xue et al., 2019) [1]. Herein, we report a complementary qPCR study that investigated the impact of the pIC, bacterin and dietary CpG on the expression of immune-relevant mRNAs (n = 31) using the same samples as in the previous study (Xue et al., 2019) [1]. Twenty-six of these genes were predicted target transcripts of the pIC- and/or bacterin-responsive miRNAs identified in the earlier study. The current data showed that pIC and/or bacterin stimulations significantly modulated the majority of the qPCR-analyzed genes involved in various immune pathways. Some genes responded to both stimulations (e.g. tnfa, il10rb, ifng, irf9, cxcr3, campb) while others appeared to be stimulation specific [e.g. irf3, irf7a, il1r1, mxa, mapk3 (pIC only); clra (bacterin only)]. A. salmonicida bacterin stimulation produced a strong inflammatory response (e.g. higher expression of il1b, il8a and tnfa), while salmon stimulated with pIC showed robust interferon responses (both type I and II). Furthermore, the current data indicated significant down-regulation of immune-relevant transcripts (e.g. tlr9, irf5, il1r1, hsp90ab1, itgb2) by dietary immunostimulant CpG, especially among pre-injection and PBS-injected fish. Together with our prior miRNA study, the present research provided complementary information on Atlantic salmon anti-viral and anti-bacterial immune responses and on how dietary CpG may modulate these responses.

Insights into the functional properties of a natural free amino acid mix: Effect on growth performance, nutrient metabolism, and immune response in a carnivorous fish, Asian seabass (Lates calcarifer).

Dietary supplements containing a functional feed additive have been shown to be beneficial to fish and shellfish aquaculture. However, the functional properties of aquafeed formulations have rarely been reported in fish. This study aimed to investigate the effects of natural free amino acid mix (FAAM) supplementation as a functional solution on the growth performance and nutrient utilization in a carnivorous fish, Asian seabass (Lates calcarifer). Five isonitrogenous and isolipidic diets were prepared with graded supplementation levels of FAAM at 0 % (control group), 0.25 %, 0.50 %, 0.75 %, and 1.0 %, denoted as FAAM0, FAAM0.25, FAAM0.5, FAAM0.75, and FAAM1.0, respectively. The experimental fish were fed different dietary FAAM supplementations to apparent satiation twice daily for eight weeks. Significant improvements were observed in the growth performance of fish among the five groups (P < 0.05). Fish fed with FAAM0.75 displayed significantly increased activities of lysozyme, myeloperoxidase, catalase, and glutathione peroxidase (P < 0.05). The activities of digestive enzymes, including amylase, protease, and lipase, were enhanced by the supplementation of FAAM in the feed (P < 0.05), especially for the groups that contained more than 0.5 % FAAM in the feed. Furthermore, the morphological profile of the intestinal tract, including the mucosal fold height, width, thickness, and goblet cell, increased in fish fed with FAAM at 1.0 % (P < 0.05). Moreover, FAAM supplementation in diets not only modulated the expression of immune-related genes (glutathione peroxidase (GPx), complement (C)3, C4, and C-reactive protein) in the liver but also positively impacted the growth-ralated genes, including growth hormone (GH), GH receptor (GHR), insulin-like growth factor I (IGF-I), and IGF-II. In addition, the amounts of monounsaturated fatty acids (mainly oleic acid (C18:1n9c)) and polyunsaturated fatty acids-especially γ-linolenic acid (C18:3 n6) and α-linolenic acid (C18:3n3)-increased in fish fed with diets containing FAAMs (P < 0.05). Interestingly, the diets supplemented with FAAMs also had a positive effect on the economic indices in terms of revenue-to-cost ratios. These findings provide a scientific basis for the application of FAAMs as a functional solution that can be used in feed formulations for Asian seabass.

Advances in the concept of functional foods and feeds: applications of cinnamon and turmeric as functional enrichment ingredients.

Spices are a rich source of vitamins, polyphenols, proteins, dietary fiber, and minerals such as calcium, magnesium, iron, and zinc, all of which play an important role in biological functions. Since ancient times, spices have been used in our kitchen as a food coloring agent. Spices like cinnamon and turmeric allegedly contain various functional ingredients, such as phenolic and volatile compounds. Therefore, this review aims to summarize the current knowledge about the nutritional profiles of cinnamon and turmeric, as well as to analyze the clinical studies on their extracts and essential oils in animals and humans. Furthermore, their enrichment applications for food products and animal feed have also been investigated in terms of safety and toxicity. Numerous studies have shown that cinnamon and turmeric have various health benefits, including the reduction of insulin resistance and insulin signaling pathways in diabetic patients, the reduction of inflammatory biomarkers, and the maintenance of gut microflora in both animals and humans. The food and animal feed industries have taken notice of these health benefits and have begun to promote cinnamon and turmeric as healthy foods. This has resulted in the development of new food products and animal feeds that contain cinnamon and turmeric as primary ingredients, which have been deemed an effective means of promoting cinnamon and turmeric's health benefits.

The antibacterial activity and immunomodulatory effect of naturally synthesized chitosan and silver nanoparticles against Pseudomonas fluorescence infection in Nile tilapia (Oreochromis niloticus): An in vivo study.

A sixty-day feeding trial was conducted to assess the effects of silver nanoparticles (AgNPs) and chitosan nanoparticles (CNPs) on the growth and immunity of Nile tilapia (Oreochromis niloticus), compared with the control group. CNPs and AgNPs were green synthesized and added to a control diet (30% crude protein) at levels of 2.0 g CNPs/kg diet and 1.0 mg AgNPs/kg diet. One hundred and eighty fish (101 ± 3.98 g) were randomly distributed into nine fiberglass tanks (200 cm × 200 cm x 100 cm, twenty fish each) to represent three equal groups (60 fish per group). After one and two months of the feeding trial, parameters of water quality, growth indices, hematology, and liver and kidney biomarkers were evaluated. At the end of the experiment, 10 fish from each group were challenged experimentally via the intraperitoneal injection with Pseudomonas fluorescence and fish mortality was observed for further ten days. Then, specimens from the liver, kidney, spleen, and anterior intestine were examined to assess the histopathological alterations. Incorporating a 2.0 g CNPs/kg diet was a promising growth enhancer; however, a 1.0 mg AgNPs/kg diet had no effects on tilapia performance. Furthermore, AgNPs appeared to reduce water pollution, leading to water filtration via decreasing both total dissolved solids (TDS) and electrical conductivity (EC). A significant role of AgNPs in improving tilapia's erythrogram (RBCs number and Hb concentration) was evident. Compared with the control group, both groups of CNPs and AgNPs improved non-specific immune parameters and showed defense effects against P. fluorescence. The fish mortality after P. fluorescence infection in CNPs and AgNPs-fed fish groups revealed significant decreases (P < 0.05) of 10% and 25%; respectively, while the control group exhibited a mortality rate of 40%. The current investigation evoked that using dietary CNPs (2.0 g/kg feed) as an antibacterial agent against P. fluorescence infection in Nile tilapia culture was better than dietary AgNPs (1.0 mg/kg diet) which, induced cells inflammation causing tissues necrosis.

Dietary supplementation of multi-strain probiotic in male rainbow trout (Oncorhynchus mykiss) broodstock: Effects on feed efficiency, hemato-biochemical parameters, immune response, and semen quality.

The present study aimed to determine the effects of dietary probiotic supplementation on feed efficiency, physiological parameters, and semen quality of male rainbow trout (Oncorhynchus mykiss) broodstock. For this purpose, a total of 48 breeders with an average initial weight of 1366.1 ± 33.8 g were divided into 4 groups and 3 replicates. Fish were fed with diets containing 0 (control), 1 × 109 (P1), 2 × 109 (P2), and 4 × 109 (P3) CFU multi-strain probiotic kg-1 diet for 8 weeks. According to the results, P2 treatment significantly enhanced body weight increase, specific growth rate, and protein efficiency ratio and decreased feed conversion ratio. Moreover, the highest values of red blood cells count, hemoglobin, and hematocrit values were observed in P2 treatment (P < 0.05). The lowest levels of glucose, cholesterol, and triglyceride were found in P1, P2, and P3 treatments, respectively. Also, the highest levels of total protein and albumin were obtained in P2 and P1 treatments (P < 0.05). Based on the results, plasma enzymes contents were significantly decreased in P2 and P3 treatments. In terms of immune parameters, the complement component 3, complement component 4, and immunoglobulin M levels were increased in all probiotic-fed treatments (P < 0.05). For spermatological features, the highest spermatocrit value, sperm concentration, and motility time were observed in the P2 treatment (P < 0.05). Consequently, we conclude that multi-strain probiotics can be used as functional feed additives in male rainbow trout broodstock to enhance semen quality, improve physiological responses, and better feed efficiency.

Potential effects of dietary seaweeds mixture on the growth performance, antioxidant status, immunity response, and resistance of striped catfish (Pangasianodon hypophthalmus) against Aeromonas hydrophila infection.

Striped catfish (Pangasianodon hypophthalmus) is one of the important fish species in the world, which is widely cultured in several countries worldwide including Egypt. Aeromonas hydrophila infection showed a high prevalence in the farmed striped catfish negatively affecting its production causing economic loss. Therefore, the finding of functional feed supplements with antimicrobial activity is urgently needed. The use of marine seaweeds as feed supplements has produced satisfactory results as antibacterial agents. Accordingly, the present study was designed to evaluate the dietary effects of seaweeds mixture (Ulva lactuca, Jania rubens, and Pterocladia capillacea) extract (SME) on the growth performance, haemato-chemical parameters, antioxidant capacity, immune indices, and resistance of striped catfish fry against A. hydrophila infection. Four isonitrogenous (30% crude protein) diets were prepared to contain 0.0% (control), 1%, 2%, and 3% SME and were fed to striped catfish fry (1.7 ± 0.1 g) up to apparent satiety three times a day for 60 days. After the feeding trial, fish of each treatment were intraperitoneally injected with a virulent A. hydrophila strain and the relative percentage of survival (RPS) was recorded. The growth-stimulating activity of dietary SME was observed especially at 2% and 3% SME treatments. Similarly, haematological parameters (red blood cells, haemoglobin, haematocrit, and white blood cells) and serum protein profile (total protein, albumin, and globulin) were significantly (P < 0.05) improved, meanwhile serum alanine and aspartate aminotransferases were significantly (P < 0.05) decreased in fish fed with 2 and 3% SME diets. Serum creatinine and uric acid levels revealed no significant (P > 0.05) differences among the experimental groups. Hepatic malondialdehyde and serum nitrous acid levels were significantly decreased; meanwhile hepatic superoxide dismutase, catalase, and glutathione peroxidase levels were significantly increased in all SME-treated groups than the control one. Also, SME supplementations significantly increased the serum lysozyme, total immunoglobulin, and complement C3 activities with highest values at 2% and 3% SME treatments. Interestingly, after bacterial challenge, fish fed SME-supplemented diet were more resistant with the lowest fish mortality (15%) at the 3% SME diet, which exhibit the highest RPS (81.9%). Meanwhile the control group showed highest mortality (85%). According to the regression fitting curve, the current study recommends using the dietary SME with optimum levels of 2.25-2.5% to improve the growth performance, welfare status, and resistance of striped catfish fry against A. hydrophila infection.

Dietary methionine supplementation improves the European seabass (Dicentrarchus labrax) immune status following long-term feeding on fishmeal-free diets.

Methionine is a limiting amino acid (AA) in fish diets, particularly in those containing high levels of plant protein (PP), and is key in the immune system. Accordingly, outcome on the fish immune mechanisms of methionine-deficient and methionine-supplemented diets within the context of 0 % fishmeal formulation, after a short and prolonged feeding period, was studied in European seabass (Dicentrarchus labrax). For this, seabass juveniles were fed a (i) fishmeal-free diet, meeting AA requirements, but deficient in methionine (MET0·65); (ii) as control, the MET0·65 supplemented with l-methionine at 0·22 % of feed weight (CTRL); (iii) two diets, identical to MET0·65 but supplemented at 0·63 and 0·88 % of feed weight of l-methionine (MET1·25 and MET1·5, respectively); and (iv) a fishmeal-based diet (FM), as positive control. After 2 and 12 weeks of feeding, blood and plasma were sampled for leucocyte counting and humoral parameter assays and head-kidney collected for gene expression. After 2 weeks of feeding, a fishmeal-free diet supplemented with methionine led to changes in the expression of methionine- and leucocyte-related genes. A methionine immune-enhancer role was more evident after 12 weeks with an increased neutrophil percentage and a decreased expression of apoptotic genes, possibly indicating an enhancement of fish immunity by methionine dietary supplementation. Furthermore, even though CTRL and FM present similar methionine content, CTRL presented a reduced expression of several immune-related genes indicating that in a practical PP-based diet scenario, the requirement level of methionine for an optimal immune status could be higher.

Effect of a functional feed additive on mitigation of experimentally induced gilthead sea bream Sparus aurata enteromyxosis.

In gilthead sea bream Sparus aurata, infection by Enteromyxum leei produces a cachectic syndrome with anorexia, weight loss, severe epaxial muscle atrophy and, eventually, death. Currently, there are neither vaccines nor effective prescription medicines to control this infection. Nutraceutical approaches are raising interest in the aquaculture industry, responding to the lack of therapeutic tools for the management of insidious chronic losses due to parasites. In this study, the effect of a commercially available health-promoting feed additive (SANACORE® GM) at 2 different doses was tested in comparison with a basal diet without the additive during a laboratory-controlled challenge with E. leei. Group performance and biometrical values were monitored, and an in-depth parasitological diagnosis, quantification of parasite loads and histopathological examination were carried out at the end of the trial. Supplemented diets mitigated the anorexia and growth arrestment observed in challenged fish fed the basal diet. This mitigation was maximum in the highest dose group, whose growth performance was not different from that of unchallenged controls. Treated groups also presented lower prevalence of infection and a lower parasite load, although the differences in the mean intensity of infection were not statistically significant. Although the decrease in parasite levels was similar with both doses of additive tested, the pathogeny of the infection was mostly suppressed with the higher dose, while only mitigated with the lower dose. The mechanisms involved in the effects obtained remain to be investigated, but the results point to a modulation of the immunopathological response to the infection.

Effect of yeast (Xanthophyllomyces dendrorhous) and plant (Saint John's wort, lemon balm, and rosemary) extract based functional diets on antioxidant and immune status of Atlantic salmon (Salmo salar) subjected to crowding stress.

Salmon farming may face stress due to the intensive culture conditions with negative impacts on overall performance. In this aspect, functional feed improves not only the basic nutritional requirements but also the health status and fish growth. However, to date no studies have been carried out to evaluate the effect of functional diets in salmon subjected to crowding stress. Thus, the aim of this study was to evaluate the effect of yeast extract (Xanthophyllomyces dendrorhous; diet A) and the combination of plant extracts (common Saint John's wort, lemon balm, and rosemary; diet B) on the antioxidant and immune status of Atlantic salmon grown under normal cultured conditions and then subjected to crowding stress. Fish were fed with functional diets during 30 days (12 kg/m3) and then subjected to crowding stress (20 kg/m3) for 10 days. The lipid peroxidation in gut showed that both diets induced a marked decrease on oxidative damage when fish were subjected to crowding stress. The protein carbonylation in muscle displayed at day 30 a marked decrease in both functional diets that was more marked on the stress condition. The expression of immune markers (IFNγ, CD4, IL-10, TGF-ß, IgMmb, IgMsec, T-Bet, and GATA-3) indicated the upregulation of those associated to humoral-like response (CD4, IL-10, GATA-3) when fish were subjected to crowding stress. These results were confirmed with the expression of secreted IgM. Altogether, these functional diets improved the antioxidant status and increased the expression of genes related to Th2-like response suggesting a protective role on fish subjected to crowding stress.

Selected fruit pomaces: Nutritional profile, health benefits, and applications in functional foods and feeds.

The utmost objective of every nation is to achieve zero hunger and ensure the health and well-being of its population. However, in impoverished nations, particularly in rural areas, such issues persist on a daily basis. Currently, there is a growing demand for fruit consumption due to their potential health benefits. Surprisingly, their most prevalent by-product is pomace, which is produced in millions of tonnes and is usually discarded as waste after processing or consumption. Even food produced with these kinds of raw resources can contribute to the objective of eradicating world hunger. Owing to these advantages, scientists have begun evaluating the nutritional content of various fruit pomace varieties as well as the chemical composition in different bioactive constituents, which have significant health benefits and can be used to formulate a variety of food products with notable nutraceutical and functional potential. So, the purpose of this review is to understand the existing familiarity of nutritional and phytochemical composition of selected fruit pomaces, those derived from pineapple, orange, grape, apple, and tomato. Furthermore, this article covers pre-clinical and clinical investigations conducted on the selected fruit pomace extracts and/or powder forms and its incorporation into food products and animal feed. Adding fruit pomaces reduces the glycemic index, increases the fibre content and total polyphenolic contents, and reduces the cooking loss, etc. In animal feeds, incorporating fruit pomaces improves the antioxidant enzyme activities, humoral immune system, and growth performance and reduces methane emission.

Salinity change evokes stress and immune responses in Atlantic salmon with microalgae showing limited potential for dietary mitigation.

Smoltification was found to impact both immune and stress responses of farmed Atlantic salmon (Salmo salar), but little is known about how salinity change affects salmon months after completed smoltification. Here, we examined (1) the effect of salinity change from brackish water to seawater on the stress and immune responses in Atlantic salmon and (2) evaluated if functional diets enriched with microalgae can mitigate stress- and immune-related changes. Groups of Atlantic salmon were fed for 8 weeks with different microalgae-enriched diets in brackish water and were then transferred into seawater. Samples of the head kidney, gill, liver and plasma were taken before seawater transfer (SWT), 20 h after SWT, and 2 weeks after SWT for gene-expression analysis, plasma biochemistry and protein quantification. The salmon showed full osmoregulatory ability upon transfer to seawater reflected by high nkaα1b levels in the gill and tight plasma ion regulation. In the gill, one-third of 44 investigated genes were reduced at either 20 h or 2 weeks in seawater, including genes involved in cytokine signaling (il1b) and antiviral defense (isg15, rsad2, ifit5). In contrast, an acute response after 20 h in SW was apparent in the head kidney reflected by increased plasma stress indicators and induced expression of genes involved in acute-phase response (drtp1), antimicrobial defense (camp) and stress response (hspa5). However, after 2 weeks in seawater, the expression of antiviral genes (isg15, rsad2, znfx1) was reduced in the head kidney. Few genes (camp, clra, c1ql2) in the gill were downregulated by a diet with 8% inclusion of Athrospira platensis. The results of the present study indicate that salinity change months after smoltification evokes molecular stress- and immune responses in Atlantic salmon. However, microalgae-enriched functional diets seem to have only limited potential to mitigate the related changes.

Lupin Seed Supplementation as a Functional Feed Additive: In Vitro Ruminal Gas, Methane and Carbon Dioxide Production, Fermentation Kinetics, and Nutrient Degradability.

The inevitable enteric gas emission from ruminants is considered a modern-day problem from an environmental perspective. Addressing this problem requires nutritional approaches such as the use of phytogenic additives in ruminant diets. In this regard, lupin seed (LS) can be a useful additive due to its phytochemical constituents. Therefore, this study investigated the effects of lupin (Lupinus angustifolius) seed supplementation as a functional and sustainable feed additive in sheep diet (50:50 concentrate-to-forage ratio) on in vitro gas production (GP; mL/g DM), methane (CH4; mL/g DM) and carbon dioxide (CO2; mL/g DM) emissions, fermentation parameters, and nutrient degradability (g/kg DM incubated). Gas production and CH4 were measured per gram of incubated dry matter (DM), degradable DM (dDM), degradable neutral detergent fiber (dNDF), and degradable acid detergent fiber (dADF). Lupin seeds were included at 0 (control), 0.5, 1.0, 1.5, and 2% of the diet. The seeds contained 3.27% essential oils (DM basis), with eucalyptol as the main phytochemical. The highest GP per gram of DM and dDM was observed (p < 0.01) with 2.0% LS supplementation level. While 1.0% LS had the highest GP per gram of dNDF, 0.5% lupin diet had the highest GP per gram of dADF. Asymptotic GP and CH4 emissions linearly and quadratically increased (p < 0.01) with increasing LS level, while lag time decreased. Despite increased CH4 production, the proportion of CH4 in total biogas was lower (p = 0.008) for LS treatments than the control, with the 0.5% LS showing the lowest CH4 proportion. Production of CO2 increased with lupin seed treatments, with 0.5% LS producing the highest proportion (p = 0.027). Degradability of DM, NDF, and ADF was greater (p < 0.01) for the high LS supplementation level, while 0.5% supplementation level decreased ADF degradability. Total short-chain fatty acids, acetic acid, and propionic acid increased (p < 0.05) with LS supplementation level, leading to a reduced acetate:propionate ratio. Rumen pH decreased (p = 0.036) with LS supplementation, while ammonia-N decreased (p = 0.045) and estimated metabolizable energy increased (p < 0.001) linearly. Calculated microbial protein synthesis (p = 0.005) and gas yield (p = 0.047) increased with LS supplementation level. LS supplementation at 2.0% of diet (DM basis) increased GP and CH4 emission (mL/g DM) and enhanced nutrient degradability, suggesting its potential use as a functional feed additive for ruminants when supplemented at a 2.0% level into diet.

Dietary Chlorella vulgaris supplementation modulates health, microbiota and the response to oxidative stress of Atlantic salmon.

Microalgae are emerging as functional feed ingredients in aquaculture due to their immune-stimulating and stress-modulating properties. We investigated the potential of the microalgae Chlorella vulgaris as a feed supplement to improve the health and modulate microbiota and stress responses of Atlantic salmon. Triplicate groups of Atlantic salmon (~ 126 g) were reared in a recirculating aquaculture system (RAS) at 15 °C and received diets supplemented with 2% (CV2) or 14% (CV14) spray-dried C. vulgaris daily, 14% once weekly (CV14w), or a control diet (CD) for 8 weeks. Subsequently, all groups were exposed to an acute one-hour peracetic acid (CH3CO3H; PAA) treatment, a commonly used disinfectant in RAS. While CV14 increased feed conversion (FCR) significantly, feeding the diets CV2 and CV14w improved protein retention efficiency. CV14 significantly modulated beta-diversity in the intestinal digesta and mucosa, but this effect was already visible in fish fed CV2. Feeding CV14 and, to a lesser degree, CV2 increased the relative abundances of Paenarthrobacter and Trichococcus in the digesta and mucosa, which are able to metabolize complex carbohydrates. However, the same diets reduced the abundance of the lactic acid bacteria Lactobacillus and Weissella in the digesta and Floricoccus in the mucosa. Peracetic acid exposure induced systemic stress (increase in plasma glucose and cortisol) and a local immune response in the gill, with the most prominent upregulation of several immune- and stress-regulated genes (clra, cebpb, marco, tnfrsf14, ikba, c1ql2, drtp1) 18 h after exposure in fish fed the control diet. Fish receiving CV14 once a week showed a reduced transcriptional response to PAA exposure. Catalase protein abundance in the liver increased following exposure to PAA, while superoxide dismutase abundance in the gill and liver was increased in response to C. vulgaris inclusion before stress. Overall, the results highlight that a high (14%) inclusion rate of C. vulgaris in feed for Atlantic salmon impairs feed conversion and shifts the intestinal microbiota composition in digesta and mucosa. Weekly feeding of C. vulgaris proves a viable approach in improving protein retention and improving transcriptional resilience towards oxidative stress in increasingly intensive production systems. Thereby this study may motivate future studies on optimizing temporal feeding schedules for health-promoting aquafeeds.

The effect of peanut skins as a natural antimicrobial feed additive on ileal and cecal microbiota in broiler chickens inoculated with Salmonella enterica Enteritidis.

The consumption of poultry products contaminated with Salmonella species is one of the most common causes of Salmonella infections. In vivo studies demonstrated the potential application of peanut skins (PS) as an antimicrobial poultry feed additive to help mitigate the proliferation of Salmonella in poultry environments. Tons of PS, a waste by-product of the peanut industry, are generated and disposed in U.S. landfills annually. Peanut skins and extracts have been shown to possess antimicrobial and antioxidant properties. Hence, we aimed to determine the effect of PS as a feed additive on the gut microbiota of broilers fed a control or PS supplemented (4% inclusion) diet and inoculated with or without Salmonella enterica Enteritidis (SE). At hatch 160 male broilers were randomly assigned to 4 treatments: 1) CON-control diet without SE, 2) PS-PS diet without SE, 3) CONSE-control diet with SE, 4) PSSE-PS diet with SE. On d 3, birds from CONSE and PSSE treatments were inoculated with 4.2 × 109 CFU/mL SE. At termination (4 wk), 10 birds/treatment were euthanized and ileal and cecal contents were collected for 16S rRNA analysis using standard methodologies. Sequencing data were analyzed using QIIME2. No effect of PS or SE was observed on ileal alpha and beta diversity, while evenness, richness, number of amplicon sequence variants (ASV) and Shannon, as well as beta diversity were significantly (P < 0.05) affected in ceca. Similarly, more differentially abundant taxa between treatment groups were identified in ceca than in ileum. However, more microbiota functional changes, based on the PICRUST2 prediction, were observed in ileum. Overall, relatively minor changes in microbiota were observed during SE infection and PS treatment, suggesting that PS addition may not attenuate the SE proliferation, as shown previously, through modulation of microbiota in gastrointestinal tract. However, while further studies are warranted, these results suggest that PS may potentially serve as a functional feed additive for poultry for improvement of animal health.

The effect of dietary supplementation of Lycium barbarum leaves on the growth performance, organ indexes and intestinal microflora of rats.

This study was conducted to investigate both fruit and different levels of leaf supplementation on the growth performance, organ indices and intestinal microflora of rats. Twenty-five healthy male Sprague-Dawley rats were randomly divided into five groups. The rats in the control (NC) and positive control (PC) groups were fed by gavage a basal diet and a basal diet with 4 g/kg of L. barbarum fruit homogenate, respectively. The test (LD, MD, and HD) groups were fed basal diets with additional 2, 4, and 8 g/kg of L. barbarum leaf homogenate, respectively. The feeding period was 35 d. The result revealed that the rats in the LD group had the highest average weight gain (p < 0.05). The cardiac and renal indexes in the LD and MD groups were significantly higher than in NC group, respectively (p < 0.05). Diversity analysis revealed that adding low concentrations of L. barbarum leaf homogenates markedly reduced the Shannon index of the rats cecum (p < 0.05). The relative abundance of Verrucomicrobiota was higher in the LD group than those in other groups (p < 0.05). The relative abundance of Actinobacteriota was found significantly higher in PC group than others (p < 0.05). The relative abundance of Akkermansia in LD group was the highest (p < 0.05). The relative abundance of Romboutsia in the PC group was considerably higher than that in other groups. The relative abundance of Candidatus_Saccharimonas in the supplementation groups was appreciably lower than those found in other groups. The relative abundance of Alloprevotella was significantly lower in PC, LD, and MD groups than in NC and HD groups (p < 0.05). The relative abundance of Oscillibacter was significantly higher in HD group than in other groups (p < 0.05). Thus, L. barbarum leaf homogenate fed to rats could increase their growth performance, internal organ weights and additionally enhance the relative abundance of beneficial bacteria. Therefore, based on the obtained data in the current study, a dose of L. barbarum leaf homogenate supplemented with 2 g/kg in diet is recommended, however, further studies are required to confirm, especially in animals.

Autochthonous Bacilli and Fructooligosaccharide as Functional Feed Additives Improve Growth, Feed Utilisation, Haemato-Immunological Parameters and Disease Resistance in Rohu, Labeo rohita (Hamilton).

The effects of Bacillus spp. (7 Log CFU g-1 feed) and fructooligosaccharide (FOS, 1%) as functional feed additives, either alone or in combination, were evaluated in a study on rohu, Labeo rohita fingerlings. The fish were fed different diets for 90 days, including a control diet and diets supplemented with FOS, B. licheniformis, B. methylotrophicus or synbiotic formulations of these. The results showed that the combination of B. licheniformis and FOS significantly improved weight gain, feed utilisation and protease activity compared to the other groups. Overall, the groups supplemented with probiotics and synbiotics (B. licheniformis + FOS or B. methylotrophicus + FOS) showed improvements in haematology, serum biochemistry and immune parameters compared to the control group. After 90 days of experimental feeding, the fish were challenged with pathogenic Aeromonas hydrophila, and data on haematology, immunity and stress parameters were collected. The results indicated that the application of Bacillus spp. and FOS boosted immunity and resistance to physiological stress in the fish. The highest post-challenge survival rate was observed in fish fed a diet with B. licheniformis and FOS, indicating the potential of this particular combination of functional feed additives to enhance growth, immunity and disease resistance in L. rohita.

Effects of dietary supplementation of multi-strain probiotics on semen quality, seminal plasma compositions, and fertilization ability of rainbow trout (Oncorhynchus mykiss) broodstock spermatozoa.

The present experiment aims to study the effects of dietary multi-strain probiotics on semen quality, seminal plasma compositions, and fertilization ability of male rainbow trout. For this purpose, a total of 48 broodstocks with an average initial weight of 1366.1 ± 33.8 g were divided into 4 groups and 3 replicates. Fish were fed with diets containing 0 (control), 1 × 109 (P1), 2 × 109 (P2), and 4 × 109 (P3) CFU probiotic kg-1 diet for 12 weeks. Results showed that dietary supplementation of probiotics significantly increased plasma testosterone level, motility time of spermatozoa, sperm density, and spermatocrit value in P2 and P3 treatments and Na+ level in P2 treatment compared to the control group (P < 0.05). The activities of aspartate aminotransferase and lactate dehydrogenase had significantly decreased in the P2 treatment compared to the control group (P < 0.05). No considerable variations were observed between control fish and treatment groups (P > 0.05) in semen biochemical parameters, percentage of motile spermatozoa, osmolality, and pH of seminal plasma. Based on the results, the highest fertilization rate (97.2 ± 0.9%) and eyed egg survival (95.7 ± 1.6%) were observed in the P2 treatment, and those values showed remarkable differences with the control group (P < 0.05). The results indicated that multi-strain probiotics have potential efficacy on semen quality and fertilization ability of rainbow trout broodstock spermatozoa.

Effect of unripe banana flour as a functional feed ingredient on growth performance, internal organ relative weight and carcass traits of broilers.

BACKGROUND: Following the prohibition of in-feed antibiotics, poultry nutritionists are increasingly interested in the use of functional feed. Unripe banana flour (UBF) contains significant amounts of oligosaccharides (which may act as prebiotics) and antioxidants, making it a potential functional feed for broilers. However, research on the use of UBF as a functional feed ingredient for broilers is limited. OBJECTIVES: The study investigated the effect of UBF with or without probiotic and multienzyme on growth, internal organ weight and carcass characteristics of broilers. METHODS: A total of 392 broiler chicks were distributed into 4 groups included CONT (chicks receiving control feed), UBF (chicks receiving 5% UBF in feed), UBFPRO (5% UBF plus 0.05% probiotics) and UBFZYM (5% UBF plus 0.05% multienzyme). Data on growth performance were weekly recorded, whereas data on internal organs and carcass were collected on day 38. RESULTS: Feed conversion ratio (FCR) was lower (p < 0.05) in UBF, UBFPRO and UBFZYM than that in CONT chicks, with no significant difference in body weight, body weight gain and feed intake. There was a tendency that gizzard was higher (p = 0.08) in CONT than in UBF chicks. Also, pancreas tended (p = 0.09) to be lower in UBFZYM than in CONT birds. There was a notable effect (p < 0.05) of dietary treatments on the yellowness (b*) values of thigh meats, in which UBFPRO had lower b* values than that of CONT but did not differ from that of UBF and UBFZYM. There was no difference (p < 0.05) in carcass and commercial proportion of broilers. CONCLUSIONS: Feeding of 5% UBF with or without probiotic and multienzyme improved FCR, without negatively affecting the carcass characteristics of broilers.

Research Progress on Lycopene in Swine and Poultry Nutrition: An Update.

Oxidative stress and in-feed antibiotics restrictions have accelerated the development of natural, green, safe feed additives for swine and poultry diets. Lycopene has the greatest antioxidant potential among the carotenoids, due to its specific chemical structure. In the past decade, increasing attention has been paid to lycopene as a functional additive for swine and poultry feed. In this review, we systematically summarized the latest research progress on lycopene in swine and poultry nutrition during the past ten years (2013-2022). We primarily focused on the effects of lycopene on productivity, meat and egg quality, antioxidant function, immune function, lipid metabolism, and intestinal physiological functions. The output of this review highlights the crucial foundation of lycopene as a functional feed supplement for animal nutrition.

Exploring the Antioxidant and Genoprotective Potential of Salicornia ramosissima Incorporation in the Diet of the European Seabass (Dicentrarchus labrax).

The identification of novel feed materials as a source of functional ingredients is a topical priority in the finfish aquaculture sector. Due to the agrotechnical practices associated and phytochemical profiling, halophytes emerge as a new source of feedstuff for aquafeeds, with the potential to boost productivity and environmental sustainability. Therefore, the present study aimed to assess the potential of Salicornia ramosissima incorporation (2.5, 5, and 10%), for 2 months, in the diet of juvenile European seabass, seeking antioxidant (in the liver, gills, and blood) and genoprotective (DNA and chromosomal integrity in blood) benefits. Halophyte inclusion showed no impairments on growth performance. Moreover, a tissue-specific antioxidant improvement was apparent, namely through the GSH-related defense subsystem, but revealing multiple and complex mechanisms. A genotoxic trigger (regarded as a pro-genoprotective mechanism) was identified in the first month of supplementation. A clear protection of DNA integrity was detected in the second month, for all the supplementation levels (and the most prominent melioration at 10%). Overall, these results pointed out a functionality of S. ramosissima-supplemented diets and a promising way to improve aquaculture practices, also unraveling a complementary novel, low-value raw material, and a path to its valorization.