Quality of Oreochromis niloticus and Cynoscion virescens fillets and their by-products in flours make for inclusion in instant food products.

The production of fish flour is an alternative for better use of the raw material, although it is rarely used in instant food. Thus, the aimed of this study was to evaluate Oreochromis niloticus (Nile tilapia) and Cynoscion virescens (croaker) fillets and the elaboration of flour with filleting by-products for inclusion in food products. Carcasses and heads of the two fish species were cooked, pressed, ground, subjected to drying and re-grinding to obtain standardized flours. These carcass flours were seasoned (sweet and salted). This study was organized into two experimental tests: Test 1: Yield, physicochemical and microbiological analyzes of fillets and flours made from carcass and head of Nile tilapia and croaker; Test 2: Seasoned flours made from Nile tilapia carcasses. There was a difference in fillets yield, where the croaker demonstrated 46.56% and the Nile tilapia 32.60%. Nile tilapia fillets had higher protein content (17.08%) and lower lipid content (0.89%) compared to croaker fillets (14.21 and 4.45%). Nile tilapia backbone flour had the highest protein content (55.41%) and the croaker the highest ash (45.55%) and the lowest Nile tilapia (28.38%). The head flours had lower protein contents (39.86%). Flours produced with croaker backbone had higher levels of calcium and phosphorus (9.34 and 9.27%). However, Nile tilapia backbone flour showed higher contents of essential amino acids. These flours demonstrated a fine granulometry (0.23 to 0.56 mm). Seasoned flours demonstrated interaction between fish species and flavors for moisture, ash, carbohydrates, calcium and phosphorus. The highest protein content (29.70%) was for Nile tilapia flour sweet flours (31.28%) had higher protein content, while salted lipids (8.06%). Nile tilapia has a lower fillet yield, although with a high protein content and low lipid content. Comparing the flours made from filleting by-products, the backbone flour has better nutritional quality, with Nile tilapia being superior to that of croaker, especially in terms of protein and amino acids.

Effects of maternal methionine supplementation on the response of Japanese quail (Coturnix coturnix japonica) chicks to heat stress.

This study investigated the hypothesis that methionine supplementation of Japanese quail (Coturnix coturnix japonica) hens can reduce the effects of oxidative stress and improve the performance of the offspring exposed to heat stress during growth. For that, the quail hens were fed with three diets related to the methionine supplementation: methionine-deficient diet (Md); diet supplemented with the recommended methionine level (Met1); and diet supplemented with methionine above the recommended level (Met2). Their chicks were identified, weighed, and housed according to the maternal diet group from 1 to 14 d of age. On 15 d of age, chicks were weighed and divided into two groups: thermoneutral ambient (constant temperature of 23 °C) and intermittent heat stress ambient (daily exposure to 34 °C for 6 h). Methionine-supplemented (Met1 and Met2) hens had higher egg production, better feed conversion ratio, higher hatchability of total and fertile eggs, and offspring with higher body weight. Supplemented (Met1 and Met2) hens showed greater expression of glutathione synthase (GSS) and methionine sulfoxide reductase A (MSRA) genes, greater total antioxidant capacity, and lower lipid peroxidation in the liver. The offspring of hens fed the Met2 diet had lower death rate (1 to 14 d), higher weight on 15 d of age, weight gain, and better feed conversion ratio from 1 to 14 d of age. Among chicks reared under heat stress, the progeny of methionine-supplemented hens had higher weight on 35 d, weight gain, expression of GSS, MSRA, and thermal shock protein 70 (HSP70) genes, and total antioxidant capacity in the liver, as well as lower heterophil/lymphocyte ratio. Positive correlations between expression of glutathione peroxidase 7 (GPX7) and MSRA genes in hens and offspring were observed. Our results show that maternal methionine supplementation contributes to offspring development and performance in early stages and that, under conditions of heat stress during growth, chicks from methionine-supplemented hens respond better to hot environmental conditions than chicks from nonsupplemented hens. Supplementation of quail hens diets with methionine promoted activation of different metabolic pathways in offspring subjected to stress conditions.

The deficiency of nutrients such as methionine in the diet of birds is affecting fertility rate, egg production, egg weight, and progeny weight. In addition, the maternal environment influences gene expression through epigenetic mechanisms, where the conditions experienced by the parental generation during embryonic development can produce effects on the progeny. This study investigates how methionine supplementation in the diet of quail hens can reduce the effects of oxidative stress and improve the performance of progeny subjected to heat stress during growth. For that, the quail hens were fed with diets containing three different levels of methionine; and their chicks were created (15 on 35 d of age) into thermoneutral and/or intermittent heat stress ambient. It was observed that methionine supplementation in the quail hens had a positive effect on mortality during the initial phase and greater weight gain in the progeny growth phase. In addition, genetic inheritance was observed through the positive correlation between the expression of genes (maternal and progeny) related to oxidative stress. The results show that methionine supplementation in the maternal diet contributes to the development and performance of the progeny when subjected to heat stress during the growth phase.

Microencapsulated dietary supplementation coupled with sexual inversion improves the immune and antioxidant response of Nile tilapia larvae under stressful conditions.

In tilapia aquaculture, the cultivation of single-sex animals is extremely widespread, as it allows for the standardization of lots, in addition to improving the general performance of the animals. However, it is possible that hormonal inversion interacts with other factors, such as environmental and nutritional management, and modulates the immune response and antioxidant system of animals in a distinct manner. In order to test this hypothesis, an experiment was carried out using Nile tilapia larvae six days after hatching, divided into four experimental groups: NI (non-inverted animals), I (sexual inverted animals), NI + M (non-inverted supplemented with microencapsulated products) and I + M (sexual inverted and supplemented with microencapsulated products; half of which were subjected to transportation-related stress after 28 days of the experiment. At the end, the survival rate was evaluated; the gene expression of heat shock protein (HSP70), interleukin 1 beta (IL-1ß) and cyclooxygenase-2 (COX 2) via RT-PCR; also evaluated were the activity of catalase (CAT) and superoxide dismutase (SOD) enzymes, as well as the total antioxidant capacity by 2,2-diphenyl-1-picryl-hydrazyl (DPPH). Animals from the I and I + M groups had the highest survival rate (p < 0.001) regardless of transportation stress. The highest expressions of HSP70 were found in the NI group (p < 0.001, with and without transportation). For the IL-1ß gene, there was an increase in expression for animals belonging to groups NI and NI + M (no transportation); and NI (p < 0.0001, with transportation). Increased COX 2 expression was observed for all groups after transportation (p < 0.0001). The highest SOD activities were observed in groups I and I + M (without transportation, p = 0.0004), and I (with transportation, p < 0.0001). The transportation decreased the total antioxidant capacity of DPPH in all treatments (p < 0.001). Finally, when evaluating all of the results together, we came to a conclusion that sex inversion improves the immune response and antioxidant profile of animals under stressful conditions when associated with microencapsulated dietary supplementation.

Immune response in sexual inverted and non-inverted Nile tilapia fingerlings supplemented with organic acid and essential oil.

The development and intensification of tilapia farming depends on the manipulation of some physiological functions, such as the sexual inversion of larvae using a synthetic androgen (17α-methyltestosterone). This inversion, however, may represent a potential oxidative stress factor and cause damage to animals in the short, medium, and long term. Dietary supplementation of natural antioxidant compounds is an interesting alternative to combat such damage. To test this hypothesis, an experimental trial was carried out involving sexual inverted and non-inverted Nile tilapia fingerlings, both supplemented and not supplemented with a blend of organic acids and essential oils protected by microencapsulation. Animals were divided into four experimental groups: NI (non-inverted animals), I (sexual inverted animals), NI + M (non-inverted animals supplemented with microcapsules), and I + M (sexual inverted animals supplemented with microcapsules). Blood parameters (WBC - white blood cells; LY - lymphocytes; RBC - red blood cells; HGB - hemoglobin; HCT - hematocrit number; MCH - mean corpuscular hemoglobin; MCV - mean corpuscular volume and MCHC - mean corpuscular hemoglobin concentration), as well as oxidative stress markers (enzymatic activity of superoxide dismutase - SOD and catalase - CAT; and total antioxidant capacity - 2,2-diphenyl-1-picryl-hydrazyl (DPPH)) and gene expression (heat shock protein 70 kDa - HSP70) were evaluated. The HGB (p < 0.001) and HCT (p = 0.005) parameters were reduced beyond the recommended limits for the animals in group I. The MCV varied statistically between the groups (p < 0.001). However, all values were within the recommended range for the species, jointly indicating normocytic anemia in group I fingerlings at the time of collection. The activity of CAT and SOD, as well as DPPH differed statistically between the experimental groups (p < 0.001), with the lowest SOD and CAT activity, as well as the highest DPPH registered in animals supplemented with microcapsules. The expression of HSP70 was lower in I + MI animals (p < 0.001). The synergistic evaluation of the results indicates that animals sexual inverted during the larval stage have a lower total antioxidant capacity in the fingerling stage, which reflects a worsening in hematological and enzymatic parameters related to immunity; and that dietary supplementation with blend of organic acids and essential oils protected by microencapsulation is sufficient to improve the immunological response both in sexual inverted and non-inverted fingerlings.

Fatty Acid Incorporation in the Muscle, Oxidative Markers, Lipid Peroxidation and PPAR-α and SREBP-2 Expression of Zebrafish Fed Linseed Oil and Clove Leaf Essential Oil.

The objective of this study is to assess, in zebrafish, the effects of combining linseed oil (LO) and clove leaf essential oil (CLEO) on the incorporation of fatty acids in the muscle, oxidative markers, lipid peroxidation and expression of the PPAR-α (Peroxisome Proliferator-Activated Receptor-α) and the SREBP-2 (Sterol Regulatory Element Binding Protein-2) genes. Six diets were prepared, containing combinations of LO (3, 6 and 9%) and CLEO (0.5 and 1%): 3% LO + 0.5% CLEO; 3% LO + 1% CLEO; 6% LO + 0.5% CLEO; 6% LO + 1% CLEO; 9% LO + 0.5% CLEO; 9% LO + 1% CLEO. Results showed increase in the incorporation of n-3 fatty acids in the muscle concomitantly with the addition of LO and CLEO. The activities of superoxide dismutase and catalase were reduced and the glutathione content had increased. Lipid peroxidation was lower in the treatment with 1% CLEO, regardless of LO content. The expression of the PPAR-α and the SREBP-2 genes was higher in animals fed 9% LO + 0.5% CLEO. Therefore, for a greater incorporation and protection against the oxidative damages of n-3 fatty acids, a combined use of 9% LO with 0.5% CLEO is recommended for zebrafish.

Expression of the PEPT1, CAT, SOD2 and GPX1 genes in the zebrafish intestine supplemented with methionine dipeptide under predation risk.

This study evaluated the effect of methionine supplementation, predation risk and their interaction on gut histology, whole-body cortisol levels, and intestinal gene expression in zebrafish. A total of 360 one-year-old animals were maintained under two environmental conditions and fed diets containing different methionine sources. Fish were fed either a control diet (CTL, without methionine supplementation), a diet supplemented with dl-methionine (DLM), or a diet supplemented with methionine dipeptide (MM) in the absence (AP) of a predator or in the presence of the predator (PP) for 48 h or 20 days. Predator-induced stress for 20 days resulted in lower body weight. Zebrafish fed methionine-supplemented diets had higher weight gain than control fish. We found no effect of predation stress or methionine supplementation on cortisol level. Predation risk and methionine supplementation showed no interaction effect on dipeptide transporter gene expression. After 48 h of predation pressure, zebrafish had higher mRNA expression of SOD2, CAT and GPX1 in the gut. After 20 days of exposure to the predator, zebrafish fed methionine-supplemented diets had lower expression of GPX1, SOD2 and CAT than those diet CTL. Methionine dipeptide and free methionine supplementation improved growth, intestinal health and survivability of zebrafish both conditions.

Dietary supplementation with free methionine or methionine dipeptide improves environment intestinal of broilers challenged with Eimeria spp.

This study examined the influence of a diet enriched with free methionine (dl-Met) or methionine dipeptide (dl-MMet) on the intestinal health of Eimeria-challenged (EC) and unchallenged (UC) broilers. A non-supplemented, methionine-deficient diet (NS) was used as control. Treatments were arranged in a 2 × 3 factorial completely randomized design with eight replications. Broilers in the EC group were infected with sporulated oocysts of Eimeria spp. (E. acervulina, E. maxima, E. praecox, and E. mitis) at 14 d of age. Performance analysis, light and electron microscopy of the jejunum, analysis of genes related to apoptosis and cell proliferation in the jejunum, and blood tests were performed at 6 days post-inoculation (dpi). EC broilers had poorer performance than UC broilers, regardless of diet (P < 0.001). Broilers fed the dl-Met diet had greater weight gain (P = 0.004) and lower feed conversion ratio (P = 0.019) than broilers fed other diets. Jejunal sections from EC broilers fed the NS diet showed short (P = 0.001) and wide villi (P < 0.001) with increased crypt depth (P < 0.001) and reduced villus / crypt ratio (P = 0.001), jejunal absorptive surface area (P < 0.001), number of neutral goblet cells (Eimeria challenge: P = 0.048; diet P = 0.016), and mucin 2 (MUC2) gene expression (P = 0.018). EC birds fed the dl-MMet diet had higher enterocyte height (P < 0.001). Birds fed the dl-MMet diet had low lamina propria width (P = 0.009). UC broilers fed the dl-Met diet had the highest number of acidic goblet cells (P = 0.005), whereas EC broilers assigned the dl-MMet diet showed the highest number of intraepithelial lymphocytes (P = 0.033). Reduced expression of caspase-3 (CASP3) (P = 0.005), B-cell lymphoma 2 (BCL2) (P < 0.001), mechanistic target of rapamycin (MTOR) (P < 0.001), and ribosomal protein S6 kinase B1 (RPS6KB1) (P < 0.001) genes was observed in EC animals. MTOR expression levels were highest in birds fed the dl-MMet diet (P = 0.004). Plasma activities of aspartate aminotransferase (AST) was influenced by both diet (P = 0.002) and Eimeria challenge (P = 0.005), with EC broilers assigned the NS diet showing the highest levels. EC broilers fed the NS diet had higher creatine kinase (CK) activity (P = 0.049). EC broilers had lower plasma uric acid (P = 0.004) and higher serum mucoproteins level (P < 0.001). These results indicate that methionine dipeptide supplementation is able to mitigate the harmful intestinal effects of Eimeria spp. in broilers.

Expression of genes related to antioxidant activity in Nile tilapia kept under salinity stress and fed diets containing different levels of vitamin C.

The aim of this study was to examine whether (1) severe changes in salinity produced increased stress, and (2) vitamin C supplementation might reduce the observed damage in Nile tilapia. The parameters measured included condition factor, survival rate, and gene expression of catalase (CAT), heat shock protein 70 (HSP70), glutathione reductase (GSR), glutathione synthase (GSS), and glutathione peroxidase (GPx). The investigation was conducted with 160 Nile tilapia divided into four treatment groups: freshwater; 7 or 21 parts per thousand (‰) salinity, all fed a basal diet; as well as a fourth treatment group consisting of fish kept at 21‰ salinity fed a diet supplemented with vitamin C (1500 mg/kg). For gene expression analysis, liver samples were collected after 24 h or after 14 d. After 24 h, fish raised in 21‰ salinity and fed with the diet supplemented with vitamin C showed similar GPx expression as the control freshwater group. GSS expression in 21‰ salinity was similar to fish exposed to 7‰ salinity. Nile tilapia exposed to 21‰ salinity without vitamin C supplementation exhibited the highest HSP70 gene expression levels after 24 h. After 14-dtreatment, the lowest survival rate was observed in the 21‰ salinity group. After 14 d, the highest expression of GPx and GSR levels was detected in fish in the 21‰ salinity group that received vitamin C. Data indicate that vitamin C supplementation enhanced the expression of genes related to antioxidant capacity in Nile tilapia exposed to higher salinity, thereby increasing protection against the oxidative effects induced by high water salinity..

The role of cinnamon as a modulator of the expression of genes related to antioxidant activity and lipid metabolism of laying quails.

Since cinnamon has vitamins and minerals in addition to antioxidants compounds in its chemical composition studies have shown the potential of cinnamon supplementation on some important characteristics in the performance of birds. Thus, this study was conducted under the hypothesis that the inclusion of cinnamon in the laying quail diet could influence the performance of the birds through the expression of genes related to antioxidant activity and lipid metabolism. To test this hypothesis, 144 Japanese quail (Coturnix japonica) with an initial age of 18 weeks and average weight of 133g were distributed in a completely randomized design with two treatments: no cinnamon supplementation (NCS-control group) and with supplementation of 9g/kg of cinnamon powder (CPS). The experiment lasted for 84 days. At the end of the experimental period, six animals from each treatment were euthanized by cervical dislocation, blood was collected and organs weighed. Liver tissue was collected for gene expression and biochemical analyses. We observed a significant effect of cinnamon inclusion on the weight of the pancreas (P = 0.0418), intestine (P = 0.0209) and ovary (P = 0.0389). Lower weights of the pancreas and intestine, and a higher ovary weight was observed in birds receiving the CPS diet. Quails fed with cinnamon supplementation also had better feed conversion per egg mass (2.426 g /g, P = 0.0126), and higher triglyceride (1516.60 mg/dL, P = 0.0207), uric acid (7.40 mg/dL, P = 0.0003) and VLDL (300.40 mg/dL, P = 0.0252) contents. A decreased content of thiobarbituric acid reactive substances (TBARS) and lower catalase activity was observed in the liver of quails from the CPS diet (0.086 nmoles/mg PTN, and 2.304 H2O2/min/mg PTN, respectively). Quails from the CPS group presented significantly greater expression of FAS (fatty acid synthase, 36,03 AU), ACC (Acetyl-CoA Carboxylase, 31.33 AU), APOAI (apolipoprotein A-I, 803,9 AU), ESR2 (estrogen receptor 2, 0.73 AU) SOD (superoxide dismutase, 4,933.9 AU) and GPx7 (glutathione peroxidase 7, 9.756 AU) than quails from the control group. These results allow us to suggest that cinnamon powder supplementation in the diet of laying quails can promote balance in the metabolism and better performance through the modulation of antioxidant activity and the expression of genes related to lipid metabolism.

Effects of methionine supplementation on the expression of protein deposition-related genes in acute heat stress-exposed broilers.

The objective of this study was to evaluate the effect of heat stress and methionine supplementation on the gene expression of insulin-like growth factor I (IGF-I), growth hormone receptor (GHR), phosphatidylinositol 3-kinase, and regulatory 1 (PI3KR1) in the liver, as well as the expression of the atrogin 1 and cathepsin L2 (CTSL2) genes in the breast muscle of broilers. Broilers from 1-21 and 22-42 days of age were divided into three treatments related to methionine supplementation as follows: without methionine supplementation (MD), recommended level of methionine (DL1), and excess supplementation of methionine (DL2). The animals were either maintained at a thermal comfort temperature or exposed to heat stress (HS) (38°C for 24 hours, starting on day 20 or day 41 for experiments 1 and 2, respectively). The heat stress increased the body temperature at both ages. Starter period: The HS animals presented increased plasma creatinine content (P<0.0001) and the highest CTSL2 gene expression (P<0.0001). The methionine supplementation increased the IGF-I (P = 0.0144) and GHR (P = 0.0011) gene expression and decreased the CTSL2 (P = 0.0004) and atrogin 1 (P = 0.0012) gene expression. Grower period: Significant effects for the interaction between supplementation and environment were observed for GHR (P = 0.0252) and CTSL2 (P = 0.0011) gene expression. The highest GHR expression was observed in animals that remained in thermal comfort on the DL2 diet, and the lowest expression occurred in the HS animals fed the MD diet. For CTSL2, the HS animals fed the MD diet presented the highest CTSL2 gene expression, and the lowest expression was observed in the animals maintained at thermal comfort on DL1 and DL2 diets. Only methionine supplementation had effect on atrogin-1 gene expression (P<0.0001), with higher methionine content in the diet lower atrogin-1 gene expression was observed. Our results suggest that heat stress induces greater protein degradation and that methionine supplementation could induce protein deposition because methionine increased the expression of genes related to protein synthesis and decreased the expression of genes related to protein breakdown.