Addition of açai oil during the close-up dry period of Holstein cows improves colostrum quality and immune responses of their calves.

This study evaluated of the effects of açai oil during the close-up dry period of Holstein cows on colostrum quality, as well as on the immune and antioxidant responses of their calves. Sixteen multiparous cows were assigned randomly to two treatments: 1) CONTROL (n = 8) - 4.48% of soybean oil/concentrate; 2) AÇAI (n =8) - 4.48% of açai oil/concentrate. Cows fed with açai oil had greater (P≤0.04) colostrum concentrations of immunoglobulins (Ig) G (1st and 2nd milking), IgG heavy chains, IgA (only at 1st milking), alpha-lactalbumin (1st milking), total protein, and antioxidant capacity against peroxyl radicals (only at 1st milking). Cows fed with açai oil had greater serum concentrations of globulin (only on the day of calving) and total protein (only on the day of calving) (P = 0.03). Calves born of cows fed with açai oil had greater serum concentrations of total protein (only 24 and 48 h after calving) and serum concentration of IgG heavy chain (only 24 h after calving) and globulin (only 24 and 48 h after calving) (P = 0.01). These data suggest that the addition of açai oil in the cow feed during the close-up dry period boosted immunity in their calves by altering the composition of colostrum.

Protective effects of silymarin in broiler feed contaminated by mycotoxins: growth performance, meat antioxidant status, and fatty acid profiles.

The aim of this study was to determine whether the inclusion of silymarin in broiler feed was able to mitigate the adverse effects of mycotoxin on growth performance, health status, liver oxidative stress, and meat fatty acid profiles. A completely randomized design with four treatments, four repetitions, and 15 chicks per repetition was used, with the following groups: (a) feed without additives (NoMyc-NoSil), (b) feed supplemented with silymarin (NoMyc-Sil), (c) feed contaminated with mycotoxin (Myc-NoSil), and (d) feed contaminated with mycotoxin and supplemented with silymarin (Myc-Sil). Growth performance, intestinal and liver health, and meat quality were assessed. The consumption of feed contaminated with mycotoxin delayed weight gain and increased the feed conversion ratio; however, the addition of silymarin prevented these adverse effects on the chicken industry. Serum ALT activity was higher in Myc-NoSil broilers than in other groups. Intake of silymarin in healthy birds increased serum globulin concentration and reduced albumin concentration and ALT and AST serum activities compared to the Myc-NoSil group. The NoMyc-Sil birds had greater villus heights and crypt depths. Luminosity and water loss by cooking were affected by mycotoxin ingestion, changes that did not occur in the meat of birds that were supplemented with silymarin. The sum of saturated and monounsaturated fatty acids in the meat did not change among treatments, unlike the sum of polyunsaturated fatty acids higher in the meat of birds that consumed silymarin. We conclude that silymarin is a potential additive in broiler feed; it reduces impairment of growth performance at the end of the productive cycle, prevents oxidative stress, improves meat quality, and increases polyunsaturated fatty acids.

Dietary supplementation with nerolidol improves the antioxidant capacity and muscle fatty acid profile of Brycon amazonicus exposed to acute heat stress.

An increase in water temperature in the Amazon River has elicited concerns about commercially important fish species associated with food security, such as matrinxã (Brycon amazonicus). Studies have demonstrated the positive effects of diets supplemented with plant-based products that combat heat stress-induced oxidative damage. The aim of this study was to determine whether dietary supplementation with nerolidol prevents or reduces muscle oxidative damage and impairment of the fillet fatty acid profile of matrinxã exposed to heat stress. Plasma and muscle reactive oxygen species (ROS) and lipid peroxidation (LPO) levels were significantly higher in fish exposed to heat stress compared to fish not exposed to heat stress, while plasma superoxide dismutase (SOD) and glutathione peroxidase (GPx) activity was significantly lower. The total content of saturated fatty acids (SFA) in fillets was significantly higher in fish exposed to heat stress compared to fish not exposed to heat stress, while he total content of polyunsaturated fatty acids (PUFA) was significantly lower. Nerolidol prevented the increase of muscle LPO and plasma ROS and LPO levels in fish exposed to heat stress, and partially prevented the increase in muscle ROS levels. Diets containing nerolidol prevented the inhibition of muscle GPx activity in fish exposed to heat stress, and partially prevented the decrease of plasma GPx activity. The nerolidol-supplemented diet prevented the increase of fillet SFA in fish exposed to heat stress, while partially preventing the decrease of PUFA. We conclude that acute heat stress at 34 °C for 72 h causes plasma and muscular oxidative damage, and that homeoviscous adaptation to maintain membrane fluidity can represent a negative impact for fish consumers. A nerolidol diet can be considered a strategy to prevent heat stress-induced oxidative damage and impairment of muscle fatty acid profiles.

Protective role of rutin dietary supplementation mediated by purinergic signaling in spleen of silver catfish Rhamdia quelen exposed to organophosphate pesticide trichlorfon.

Evidence suggests the involvement of purinergic signaling, a mechanism mediated by extracellular nucleotides and nucleosides, with the impairment of immune and inflammatory responses in silver catfish (Rhamdia quelen) exposed to trichlorfon. Plant-derived substances have been considered potent anti-inflammatory agents due to effects on the purinergic system, such as the use of the flavonoid rutin. The aim of this study was to determine whether a diet containing rutin is able to prevent or reduce trichlorfon-induced impairment of immune responses through alteration of the purinergic pathway. Spleen adenosine triphosphate (ATP) levels were significantly higher in silver catfish exposed to 11 mg/L trichlorfon for 48 h compared to the control group, while adenosine (Ado) levels were significantly lower. Spleen ectonucleoside triphosphate diphosphohydrolase (NTPDase) activity was significantly lower in silver catfish exposed to trichlorfon compared to control group, while adenosine deaminase activity was significantly higher. Spleen metabolites of nitric oxide, interleukin-1, and IL-6 were significantly higher in silver catfish exposed to trichlorfon compared to control group. Diet with 3 mg rutin/kg diet was able to prevent all the alterations elicited by trichlorfon, except restoring spleen ATP levels. The purinergic exposure signaling is involved in impairment of immune and inflammatory responses in fish exposed to trichlorfon due to reduction in ATP hydrolyses and by an increase in Ado deamination, leading to release of pro-inflammatory mediators. Use of rutin-added diet exerted an essential role in protecting the silver catfish spleen from trichlorfon-induced impairment on immune and inflammatory responses, preventing all alterations on splenic purinergic signaling.

Topical hydrogel containing Achyrocline satureioides oily extract (free and nanocapsule) has anti-inflammatory effects and thereby minimizes irritant contact dermatitis.

Inflammatory dermatoses are prevalent worldwide, with impacts on the quality of life of patients and their families. The aim of this study was to determine the anti-inflammatory effects of Achyrocline satureioides oily extracts and nanocapsules on the skin using a mouse model of irritant contact dermatitis induced by croton oil, and a skin inflammation model induced by ultraviolet B (UVB) radiation. The mice were treated with 15 mg/ear oily extract (HG-OLAS) or nanocapsules (HG-NCAS) of A. satureioides incorporated into Carbopol® 940 hydrogels. We found that HG-OLAS and HG-NCAS formulations reduced ear edema in croton oil-induced lesions with maximum inhibitions of 54±7% and 74±3%, respectively. HG-OLAS and HG-NCAS formulations decreased ear edema induced by UVB radiation (0.5 J/cm2), with maximum inhibitions of 68±6% and 76±2% compared to the UVB radiation group, respectively. HG-OLAS and HG-NCAS modulated myeloperoxidase (MPO) activity after croton oil induction. Furthermore, croton oil and UVB radiation for 6 and 24 h, respectively, stimulated polymorphonuclear cells infiltration. The topical treatments reduced inflammatory processes, as shown by histological analysis. Together, the data suggest that topical application of A. satureioides oily extracts and nanocapsules produced antiedematogenic and anti-inflammatory effects. They constitute a compelling alternative for treatment of skin injuries.

Effects of the inclusion of açai oil in diet of prepartum Holstein cows on milk production, somatic cell counts and future lactation.

We measured the effects of açai oil in the diets of prepartum cows to evaluate health, milk production and quality. Sixteen Holstein cows were divided into two groups: SOY used as control, and AÇAI, test group. Occurred inclusion of 4% soybean or açai oils was provided in the concentrate starting at 20 days prepartum [d -20 to d 0 (partum-day)]. The AÇAI diet increased (P=0.01) milk production (d 10 and 20) and reduced somatic cell count (d 20). In milk, no effects were detected (P≥0.10) for concentration of fat, lactose or protein as well as in terms of serum concentration of calcium, albumin or triglycerides. AÇAI diet tended to increase (P=0.09) serum concentrations of total protein, glutathione transferase (d 4), and total antioxidant capacity (d 4 and 10) and increased (P≤0.05) globulin, gamma-glutamyl transferase, superoxide dismutase and glutathione peroxidase (d 4). Further, AÇAI diet reduced the serum concentration of creatine kinase (P≤0.05) (d 0, 4 and 10), reactive oxygen species (d 0 and 4) and lipoperoxidation (d 0) and tended to reduce aspartate transaminase activity (P=0.07; d 0 and 4). Açai oil in the diets in prepartum cows improved their health as well as milk production and quality.

Protective effects of diet containing rutin against trichlorfon-induced muscle bioenergetics disruption and impairment on fatty acid profile of silver catfish Rhamdia quelen.

Trichlorfon is an organophosphate insecticide that is widely used on fish farms to control parasitic infections. It has been detected in freshwater ecosystems as well as in fishery products. There is a growing body of evidence to suggest that certain feed additives may reduce or prevent pesticide-induced toxicity in fish. The aim of the present study was to determine whether acute exposure to trichlorfon would alter bioenergetic homeostasis and alter fatty acid profiles in muscles of silver catfish (Rhamdia quelen). We also sought to determine whether rutin prevents or reduces these effects. Cytosolic and mitochondrial creatine kinase (CK) and activities of complexes II-III and IV in muscle were significantly inhibited by exposure to 11 mg/L trichlorfon for 48 h compared to effects in the unexposed group. Total content of polyunsaturated fatty acids (omega-3 and omega-6) were significantly lower in muscle of silver catfish exposed to 11 mg/L trichlorfon for 48 h than in the unexposed group. Addition of 3 mg rutin/kg feed increased CK activity and prevented inhibition of complex IV activity, as well as preventing all alterations of muscle fatty acid profiles elicited by exposure to trichlorfon. No significant differences were observed between groups with respect to muscle adenylate kinase or pyruvate kinase activities, as well as total content of saturated and monounsaturated fatty acids. Our findings suggest that exposure (48 h) to 11 mg trichlorfon/L water inhibits cytosolic and mitochondrial CK activity in muscle. Trichlorfon also affects activities of complexes II-III and IV in respiratory chain, with important consequences for adenosine triphosphate production. The pesticide alters fatty acid profiles in the fish and endangers human consumers of the product. The most important finding of the present study is that inclusion of rutin improves bioenergetic homeostasis and muscle fatty acid profiles, suggesting that it reduces trichlorfon-induced muscle damage.

Dietary exposure to ochratoxin A reduces growth performance and impairs hepatic purinergic signaling in tambaqui (Colossoma macropomum).

The practice of replacing costly animal-derived proteins with more economical plant proteins has augmented the risk of mycotoxin contamination in fish feeds, including contamination with ochratoxin A (OTA). OTA is a secondary metabolite produced by molds commonly found in fish feeds that causes impairment of performance in several fish species and some hepatic biochemical alterations. However, the pathways involved in hepatic damage remain unknown and are limited to histopathological alterations. Purinergic signaling is a homeostatic system that continuously monitors the internal environment to detect injury primarily by two intercellular messengers: adenosine triphosphate (ATP) and adenosine (Ado). The objective of this study was to determine whether OTA-contaminated feed induces the release of nucleotides in the extracellular milieu, as well as whether ectoenzymes modulate ATP pro-inflammatory effects in liver of tambaqui (Colossoma macropomum). Final mean weight, weight gain (WG), and liver weight were significantly lower in tambaqui fed feeds containing 1.6 and 2.4 mg OTA/kg feed than in the control group. Liver ATP and Ado levels were significantly higher in tambaqui fed feeds containing 1.6 and 2.4 mg OTA/kg feed compared with control, while no significant difference was observed regarding adenosine diphosphate and adenosine monophosphate levels. Hepatic triphosphate diphosphohydrolase (NTPDase) activity (for ATP) was significantly greater in tambaqui fed feeds containing 1.6 and 2.4 mg OTA/kg feed compared with control, while adenosine deaminase (ADA) activity was lower. No significant difference was observed with respect to hepatic NTPDase activity (for ADP) or for 5'-nucleotidase activity. Finally, levels of liver metabolites of nitric oxide were significantly higher in tambaqui fed feeds containing 1.6 and 2.4 mg OTA/kg feed than in the control group. Based on these data, exposure to 1.6 and 2.4 mg OTA/kg feed impaired tambaqui growth performance associated with final mean weight and WG. Levels of two important intercellular messengers, ATP and Ado, increased in the extracellular space as a consequence of hepatic damage, exerting opposite immune responses. Finally, liver NTPDase and ADA activities were altered to modulate ATP and Ado levels, respectively, exerting anti-inflammatory effects to counteract OTA-induced hepatic injury.

Acute exposure to environmentally relevant concentrations of copper affects branchial and hepatic phosphoryl transfer network of Cichlasoma amazonarum: Impacts on bioenergetics homeostasis.

The toxic effects of copper (Cu) are linked to dysfunction of metabolism and depletion of adenosine triphosphate (ATP). Nevertheless, the effects related to phosphoryl transfer network, a network of enzymes to precise coupling of the ATP-production and ATP-consuming process for maintenance of bioenergetic, remain unknown. Therefore, the aim of this study was to determine whether the phosphoryl transfer network could be one pathway involved in the bioenergetic imbalance of Cichlasoma amazonarum exposed for 96 h to environmentally relevant concentrations of Cu found in Amazonia water around mines. Branchial mitochondrial creatine kinase (CK) activity was significantly lower in fish exposed to 1500 µg/L Cu than in the control group, while branchial cytosolic CK activity was significantly greater. Branchial (exposed to 750 and 1500 µg/L Cu) and hepatic (exposed to 1500 µg/L Cu) pyruvate kinase (PK) activity was significantly lower in fish exposed to Cu than in the control group. Branchial and hepatic ATP levels were significantly lower in fish exposed to 1500 µg/L than in the control group. Branchial reactive oxygen species (ROS) and lipid peroxidation (LPO) levels were significantly higher in fish exposed to 750 and 1500 µg/L Cu compared to control. Hepatic ROS and LPO levels were significantly higher in fish exposed to 1500 µg/L than in the control group. Branchial and hepatic Cu levels were significantly higher in fish exposed to 1500 µg/L compared to other groups. Exposure to 750 and 1500 µg/L Cu impairs bioenergetics homeostasis, which appears to be mediated by ROS overproduction and lipid peroxidation.

Dog food production using curcumin as antioxidant: effects of intake on animal growth, health and feed conservation.

The objectives of this study were to produce dog food containing curcumin replacing synthetic antioxidants, to evaluate its beneficial effects on animal growth and health. Curcumin (100 mg/kg) was added after the extrusion process along with the other micronutrients. The final concentration of curcumin was 32.9 mg/kg. The control feed was composed of the same ingredients without curcumin. After a storage of 6 months, feed composition and pH did not differ; however, the feed with curcumin showed lower protein oxidation, lipid peroxidation and higher total antioxidant capacity. After 2 months of feed production, 12 young Beagle dogs received either curcumin-containing food (n = 6)  or the control diet (n = 6). The animals were fed twice a day using individual kennels. Blood samples were taken on d 1, 35 and 42. During the first 30 d of the study, the animals had natural infectious diseases that were controlled with anti-protozoals and antibiotics. Greater numbers of red blood cells were observed in dogs fed with curcumin (d 35 and 45), and there were greater numbers of white blood cells as a consequence of increased neutrophils on d 42. At the end of the experiment, a significant reduction in the number of lymphocytes was observed in dogs that ingested curcumin (d 42), suggesting an anti-inflammatory effect, manifested as a decrease in globulin levels. In the final 15 d of the experiment, the animals were clinical healthy. Higher serum levels of glucose, urea, triglycerides and cholesterol were observed in dogs fed with curcumin. Curcumin increased the activity of several antioxidant enzymes in addition to non-protein thiols and the total antioxidant capacity in the serum, consequently reducing levels of oxygen reactive species. Curcumin supplementation of dogs did not favour growth or weight gain. Neverthless, it was concluded that curcumin improved animal health, with emphasis on the stimulation of the antioxidant system and evidence of an anti-inflammatory effect.

Curcumin addition in diet of laying hens under cold stress has antioxidant and antimicrobial effects and improves bird health and egg quality.

Curcumin is an herbal component with several biological properties, and we highlight here the thermal stability, antioxidant and anti-inflammatory activity. We determined whether curcumin supplementation in the diets of laying hens under cold stress and naturally infected with Escherichia coli would control infection, and would have positive effects on overall health, as well as egg production and quality. We allocated 36 chickens that had been naturally infected with E. coli into two groups, with six replicates and three chickens per repetition: control group and curcumin group (200 mg curcumin/kg). The experimental period lasted 42 days, and fecal, blood and egg samples were collected at 1, 21 and 42 days We found that feces and eggs had lower total bacterial counts, E. coli counts and total coliform counts in the curcumin group at 21 and 42 days. In fresh eggs, the brightness and yellow intensity (b+) were significantly higher in the curcumin group. In stored eggs, higher specific gravity, albumen height and lower yolk pH were observed in the curcumin group. Fresh eggs collected and stored on day 42 showed lower levels of lipid peroxidation in the curcumin group, while the total antioxidant capacity in the stored eggs was significantly higher in the curcumin group. The curcumin group showed lower total leukocyte counts as a result of lower numbers of neutrophils and lymphocytes, as well as lower levels of total protein, alkaline phosphatase and alanine aminotransferase. Lower serum lipoperoxidation at 42 days was observed in the curcumin group, probably because of the higher activity of glutathione peroxidase and glutathione transferase in other words, because of antioxidant stimulation. Taken together, our findings suggest that curcumin supplementation in laying hens under cold stress and with colibacillosis has positive effects on infection control because of antioxidant stimulation.

Dietary ochratoxin A (OTA) decreases growth performance and impairs muscle antioxidant system and meat fatty acid profiles in juvenile tambaqui (Colossoma macropomum).

Plant-based ingredients are successfully replacing fishmeal in fish feeds. However, this practice increases the risk of feed contamination by mycotoxins that reduce production and heath associated with oxidative damage. The aim of this study was to determine whether feed contaminated with environmentally relevant concentrations of ochratoxin A (OTA) causes muscle oxidative damage in tambaqui (Colossoma macropomum), and to determine whether this feed impairs fatty acid profiles in fish meat. Final mean weight, weight gain, daily weight gain, feed efficiency, and specific growth rate were significantly lower in fish fed 1.6 and 2.4 mg OTA/kg compared to those fed basal diet, while feed conversion was significantly higher in these same groups compared to the basal group. Levels of reactive oxygen species and lipid peroxidation in muscle were significantly higher in fish fed 1.6 and 2.4 mg OTA/kg compared to those in the basal group. Likewise, muscle superoxide dismutase and glutathione peroxidase activities were significantly higher in fish fed 1.6 and 2.4 mg OTA/kg then in those fed basal feed. The total content of saturated fatty acids was significantly higher, and total content of polyunsaturated fatty acids was significantly lower in fish fed 2.4 mg OTA/kg compared to those fed basal feed. Taken together, the data suggest that OTA-contaminated feed induces oxidative damage and disturbs enzymatic and non-enzymatic antioxidant responses in tambaqui meat. The perturbations of fatty acid profiles in fish meat appears to mediated by oxidative damage, representing negative impact on fish health and presenting concerns for consumers of fish.

Purine levels and purinergic signaling in plasma and spleen of Brycon amazonicus exposed to acute heat thermal stress: An attempt to regulate the immune response.

Amazon fish are vulnerable to climate change. Several lines of evidence suggest that the temperature of Amazonian rivers will increase in the coming years. Elevated temperature disturbs homeostasis and subjects fish to physiological stress; however, the effects of temperature on immunity remain poorly understood, particularly those effects involving purinergic signaling. This system fine-tunes the inflammatory and immune responses triggered by stress. Therefore, the aims of this study were to determine whether acute heat stress induces the release of nucleotides into extracellular compartment and to determine whether purinergic enzymes modulate the proinflammatory effects of adenosine triphosphate (ATP) in plasma and spleen of matrinxã (Brycon amazonicus) exposed to acute heat stress. We exposed juvenile matrinxã to four temperature regimes (28 °C as control, 30, 32 and 34 °C) for 72 h and observed the effects on purinergic signaling. Plasma cortisol levels were significantly higher in fish exposed to 34 °C than in the control group, while spleen ATP, adenosine diphosphate (ADP) and adenosine monophosphate (AMP) levels were significantly higher in this group than in controls. Activities of spleen nucleoside triphosphate diphosphohydrolase (NTPDase) and 5'-nucleotidase were significantly higher in fish exposed to 34 °C than those of the control group, while spleen interleukin-1 (IL-1) and interleukin-6 (IL-6) levels were higher in this same group than in the control group. No significant differences were observed between the groups regarding plasma parameters. Based on these data, we concluded that acute heat stress at 34 °C caused physiological stress in matrinxã, manifesting as elevated plasma cortisol levels. The most important finding is that purinergic enzymes were modulated, though not efficiently, in response to the excessive release of nucleotides into the extracellular space. In summary, the purinergic signaling pathway may be involved in the impairment of immune and inflammatory responses in matrinxã exposed acutely to 34 °C.

Purinergic signaling and gene expression of purinoceptors in the head kidney of the silver catfish Rhamdia quelen experimentally infected by Flavobacterium columnare.

The head kidney is a lymphoid immune organ that plays a key role in the immune and inflammatory responses of teleost fish. It is associated with immunoglobulin G production and differentiation of B cells. The presence of a multi-enzymatic complex found anchored in the plasma membrane makes the head kidney an important purinergic-dependent tissue. Purinergic signaling has been associated with these responses under pathological conditions via regulation of extracellular adenosine triphosphate (ATP), the main damage molecular associated pattern agent released during bacterial infections. The aim of this study was to determine whether purinergic signaling is a pathway associated with impairment of immune responses in silver catfish (Rhamdia quelen) experimentally infected by Flavobacterium columnare, as well as to evaluate the role of P2 purine receptors in this response. Triphosphate diphosphohydrolase (NTPDase) activity in the head kidney was significantly lower in silver catfish experimentally-infected F. columnare 72 h post-infection (hpi) than in the control group, while no significant difference was observed with respect NTPDase activity on adenosine diphosphate, as well as on 5'-nucleotidase and adenosine deaminase activities. Extracellular ATP levels were significantly higher in the head kidney of experimentally-infected fish than in the control group at 72 hpi. Finally, p2ry11 and p2rx3 purine receptor levels were significantly higher in experimentally-infected fish than in the control group at 72 hpi. We conclude that purinergic signaling in the head kidney of silver catfish infected by F. columnare creates a pro-inflammatory profile that may contribute to impairment of immune and inflammatory responses via reduction of ATP hydrolysis and its accumulation in the extracellular milieu, accompanied by upregulation of p2ry11 and p2rx3 purine receptors, leading to pro-inflammatory status.

Nanospheres as a technological alternative to suppress hepatic cellular damage and impaired bioenergetics caused by nerolidol in Nile tilapia (Oreochromis niloticus).

Nerolidol is a sesquiterpene found in essential oils of several plant species. It is found commonly in human and animal diets and is approved by the US Food and Drug Administration as a flavoring agent. Nevertheless, recent studies have suggested that nerolidol has potent hepatotoxic effects. Because use of plant-based products in human and animal food has expanded considerably, it is essential to develop approaches such as nanotechnology to avoid or reduce hepatic toxic effects. Therefore, the aim of the study was to determine whether nerolidol dietary supplementation elicited hepatic damage associated with impairment of energy homeostasis, as well as whether supplementation with nerolidol-loaded in nanospheres prevented hepatotoxic effects in Nile tilapia (Oreochromis niloticus). Nile tilapia were divided into five groups (A-E, n = 10 per group) with four replicates each, as follows: group A received basal feed (without supplementation); group B received feed containing 0.5 mL free nerolidol/kg; group C received feed containing 1.0 mL free nerolidol/kg; group D received feed containing 0.5 mL nanospheres nerolidol/kg; and group E received feed containing 1.0 mL nanospheres nerolidol/kg. All groups received experimental feed once a day (10% total biomass) at 2 p.m. for 60 consecutive days. Hepatic liver weight and relative liver weight were significantly lower in fish fed 1.0 mL free nerolidol/kg feed than in fish given basal diet (control group). Hepatic pyruvate kinase (1.0 mL free nerolidol/kg) and adenylate kinase (0.5 and 1.0 mL free nerolidol/kg) activities were significantly lower than in the control group, while hepatic reactive oxygen species and lipid damage levels were significantly higher. Finally, the comet assay revealed significant increases in the frequency of damage and the damage index in fish given 0.5 and 1.0 mL free nerolidol/kg in a dose-dependent manner. Nerolidol-loaded in nanospheres prevented all alterations elicited by free nerolidol. Based on these data, we concluded that dietary supplementation with free nerolidol elicited severe impairment of hepatic bioenergetics homeostasis that appeared to be mediated by excessive ROS production and lipid damage, contributing to a genotoxic effect. Dietary supplementation with nerolidol-loaded in nanospheres did not elicit hepatic damage, and therefore, should be considered as a replacement so as to limit toxicity, permitting its continued use as a dietary supplement.

Benefits of nanotechnology: Dietary supplementation with nerolidol-loaded nanospheres increases survival rates, reduces bacterial loads and prevents oxidative damage in brains of Nile tilapia experimentally infected by Streptococcus agalactiae.

Rampant and uncontrolled use of antibiotics is a major concern for aquaculture; the practice foments the emergence of resistant strains of Streptococcus agalactiae, among other negative impacts. Constituents of plant essential oils such as nerolidol are being considered as replacements for synthetic drugs to support fish nutrition and health. There is evidence to suggest that nanotechnology may enhance the efficacy of natural bioactive compounds; this is a substantial advance for the development and sustainability of aquaculture. Against the backdrop of this evidence, we aimed determine whether dietary supplementation with free nerolidol and nerolidol-loaded nanospheres would exert bactericidal effects against S. agalactiae, as well as prevent S. agalactiae-induced brain oxidative damage. In Experiment I, we measured the antimicrobial properties of dietary supplementation of nerolidol and nerolidol nanosphere in terms of mortality, longevity and relative percent survival. Fish infected with S. agalactiae fed 0.5 and 1.0 mL nerolidol nanospheres kg/diet demonstrated lower mortality and higher relative percent survival than the control group, while longevity was higher in all infected plus supplementation groups. Experiment II showed significantly lower microbial loads in brains of fish infected with S. agalactiae that were fed 1.0 mL nerolidol nanospheres kg/diet than in the control group. Brain nerolidol levels were significantly higher in uninfected as well as infected fish supplemented with nerolidol nanospheres than in fish supplemented with free nerolidol. Finally, brain reactive oxygen species and lipid peroxidation levels were higher in infected fish supplemented with basal diet compared to uninfected fish and supplemented with basal diet, and the supplementation with 1.0 mL/kg nerolidol nanospheres prevented this augmentation caused by infection. These data suggest that dietary supplementation with nerolidol nanospheres (1.0 mL/kg diet) has potent bactericidal effects in terms of augmentation of fish longevity and survival, and reduction of brain microbial loads. Also, S. agalactiae-induced brain oxidative damage that contributed to disease pathogenesis, and the dietary supplementation with nerolidol nanospheres (1.0 mL/kg diet) prevented this alteration. In summary, nanotechnology is a compelling approach to enhancing the efficacy of nerolidol, giving rise to reduction of S. agalactiae loads in fish brains.

Diphenyl diselenide modulates splenic purinergic signaling in silver catfish fed diets contaminated with fumonisin B1: An attempt to improve immune and hemostatic responses.

The aim of this study was to determine whether purinergic signaling is a pathway associated with fumonisin B1 (FB1)-induced impairment of immune and hemostatic responses. We also determined whether dietary supplementation with diphenyl diselenide (Ph2Se2) prevents or reduces these effects. Splenic nucleoside triphosphate diphosphohydrolase (NTPDase) activity for adenosine triphosphate (ATP) and adenosine diphosphate (ADP) as substrates and total blood thrombocytes counts were significant lower in silver catfish fed with FB1-contaminated diets than in fish fed with a basal diet, while splenic adenosine deaminase (ADA) activity and metabolites of nitric oxide (NOx) levels were significant higher. Also, glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were significant lower in silver catfish fed with FB1-contaminated diets than in fish fed with a basal diet. Dietary supplementation with 3 mg Ph2Se2/kg of feed effectively modulated splenic NTPDase (ATP as substrate), ADA, GPx and SOD activities, as well as NOx levels, and was partially effective in the modulation of spleen NTPDase activity (ADP as substrate) and total blood thrombocytes count. These data suggest that splenic purinergic signaling of silver catfish fed with FB1-contaminated diets generates a pro-inflammatory profile that contributes to impairment of immune and inflammatory responses, via reduction of splenic ATP hydrolysis followed possible ATP accumulation in the extracellular environment. Reduction of ADP hydrolysis associated with possible accumulation in the extracellular environment can be a pathophysiological response that restricts the hemorrhagic process elicited by FB1 intoxication. Supplementation with Ph2Se2 effectively modulated splenic enzymes associated with control of extracellular nucleotides (except ADP; that was partially modulated) and nucleosides, thereby limiting inflammatory and hemorrhagic processes.

Changes in cardiac and hepatic energetic metabolism in gerbils infected by Listeria monocytogenes.

Energy metabolism is a sensitive indicator of cellular disorders. Therefore, the objective of this study was to investigate changes in cardiac and hepatic energy metabolism during listeriosis using an experimental model. We divided gerbils into two groups: Control (n = 11) and orally Infected (n = 12) with 5 × 109 CFU/mL of Listeria monocytogenes. Euthanasia and sampling were performed on days 6 and 12 post-infection (PI). Histopathological lesions were not found in the heart; however, the liver showed pyogranuloma. In the hearts of infected animals, cytosolic creatine kinase activity was lower on day 6 and 12 PI; mitochondrial creatine kinase/pyruvate kinase (PK), and sodium potassium pump (Na+/K+-ATPase) activities were lower on day 12 PI. Hepatic PK and Na+/K+-ATPase activities were lower in the infected group on day 12 PI. Lipoperoxidation was higher in the livers and hearts of infected animals on day 12 PI, and antioxidant capacity against peroxyl radicals (ACAP) was also higher in this group. These data suggest that subclinical listeriosis alters hepatic and cardiac energy metabolism, possibly related to decreased activity of phosphotransferases and ATPase. Subsequent antioxidant responses are not sufficient to correct alterations in lipid peroxidation and bioenergetics, possibly leading to important cellular pathological mechanisms.

Branchial bioenergetics dysfunction as a relevant pathophysiological mechanism in freshwater silver catfish (Rhamdia quelen) experimentally infected with Flavobacterium columnare.

Flavobacterium columnare, the causative agent of columnaris disease, is a serious bacterial disease responsible for causing devastating mortality rates in several species of freshwater fish, leading to severe economic losses in the aquaculture industry. Notwithstanding the enormous impacts this disease can have, very little is known regarding the interaction between the host and bacterium in terms of the mortality rate of silver catfish (Rhamdia quelen), as well its linkage to gill energetic homeostasis. Therefore, we conducted independent experiments to evaluate the mortality rates caused by F. columnare in silver catfish, as well as whether columnaris disease impairs the enzymes of the phosphoryl transfer network in gills of silver catfish and the pathways involved in this inhibition. Experiment I revealed that clinical signs started to appear 72 h post-infection (hpi), manifesting as lethargy, skin necrosis, fin erosion and gill discoloration. Silver catfish began to die at 96 hpi, and 100% mortality was observed at 120 hpi. Experiment II revealed that creatine kinase (CK, cytosolic and mitochondrial) and pyruvate kinase (PK) activities were inhibited in silver catfish experimentally infected with F. columnare, while no significant difference was observed between experimental and control groups with respect to adenylate kinase activity. Activity of the branchial sodium-potassium pump (Na+, K+-ATPase) was inhibited while reactive oxygen species (ROS) and lipid peroxidation levels were higher in silver catfish experimentally infected with F. columnare than in the control group at 72 hpi. Based on these data, the impairment of CK activity elicited by F. columnare caused a disruption in branchial energetic balance, possibly reducing ATP availability in the gills and provoking impairment of Na+, K +ATPase activity. The inhibition of CK and PK activities appears to be mediated by ROS overproduction and lipid peroxidation, both of which contribute to disease pathogenesis associated with branchial tissue.

Benefits of thymol supplementation on performance, the hepatic antioxidant system, and energetic metabolism in grass carp.

Thymol is an herbal food additive used to improve animal performance. Thymol acts via its potential to enhance productive and reproductive performance, and by improving bioavailability of nutrients in fish. Nevertheless, the exact mechanisms associated with these phenomena remain poorly understood, although recent evidence has suggested the involvement of the phosphotransfer network and antioxidant status. Therefore, the aim of this study was to determine whether the improvement of the antioxidant/oxidant status and the phosphoryl transfer network may be involved in enhanced growth performance in grass carp (Ctenopharyngodon idella) fed with various levels of thymol (100, 200, and 300 mg/kg feed). Thymol-supplementation (100 mg/kg feed) produced higher body weight and weight gain for 60-day post-feeding compared to the control group. Specific growth rate was higher; while feed conversion ratio was lower in fish that consumed 100 mg of thymol/kg compared to other groups. Hepatic lactate dehydrogenase activity and lipid peroxidation levels were lower in the thymol-supplemented group (100 mg/kg feed) than in the control group, while reactive oxygen species were lower in all supplemented groups than in the control group. Hepatic superoxide dismutase (300 mg/kg feed) and glutathione peroxidase (100, 200, and 300 mg/kg feed) activities, as well as antioxidant capacity against peroxyl radicals (100 mg/kg feed) were higher in these groups than in the control group. Based on these data, we conclude that 100 mg thymol/kg dietary supplementation increased growth performance of fingerling grass carp. Finally, hepatic adenylate kinase activity was lower in the thymol supplemented group (100 mg/kg feed) than in the control group. Thymol supplementation (100 mg/kg feed) improved hepatic energy metabolism, while practically all tested concentrations of thymol enhanced hepatic antioxidant status, all of which may be pathways involved in increased growth performance in fingerling grass carp.