Impacts of intake of trichothecenes (Fusarium sporotrichioides) for dairy calves: Effects on animal growth, oxidative and inflammatory response.

The objective of the present study was to evaluate the impacts of trichothecenes (Fusarium sporotrichioides) for dairy calves on animal growth, oxidative and inflammatory responses in the presence or absence of essential oils. Twelve calves weaned at 70 days of age were divided into 2 groups: T-C (control) and T-EO (essential oils - oregano, thyme, basil and rosemary) in the period of 40 days consuming ration contaminated by trichothecenes (500 ppb). The animals in the T-EO group received a mixture of EOs via feed at a dosage of 0.75 mL per/kg of feed. Blood collections were performed on days 1, 20 and 40 for hematological and biochemical analyses; the fecal score was performed every 2 days on a scale of 1-5 and clinical examinations were performed 3 times during the experiment period. The animals were weighed at the beginning and at the end of the experiment; euthanasia of two calves per group for macroscopic and microscopic evaluation of several tissues (spleen, liver, duodenum, jejunum, ilium, cecum and colon) was performed at the end of the experiment. The calves in the T-EO group had a tendency (P = 0.07) of higher body weight when compared to the T-C. Treatment effect and treatment vs day interaction was detected for leukocytes and granulocytes variables, demonstrating a higher count of these cells in the T-EO group on both days (20 and 40), and the same behavior occurred for the distribution amplitude of erythrocytes (RDW). The enzymes alanine transferase (ALT), aspartate transferase (AST) and gamma glutamyl-transferase (GGT) showed higher serum activity in the T-C group (days 20 and 40). The levels of thiobarbituric acid reactive substances (TBARS) were lower in the serum of animals in the T-EO group. For calves in the T-EO group, glutathione S-transferase activity was higher in serum. Haptoglobulin and C-reactive protein levels were lower on days 20 and 40 in T-EO animals when compared to the T-C group. In the macroscopic and microscopic evaluations, which were collected at the end of the experiment after slaughtering the animals, liver and intestine did not show changes for the animals in the T-EO group, unlike the animals in the T-C group, which had moderately firm diffuse consistency of the liver and edema in the mesentery, as well as oxidative stress in tissues (liver, duodenum, jejunum, ileum, cecum and colon). The results concluded that the consumption of a mixture of EOs (essential oils - oregano, thyme, basil and rosemary) minimized the negative effects caused by trichothecenes in dairy calves, thus being an alternative to improving the immunological and antioxidant condition, as well as a possible adsorbent alternative.

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.

Effect of dietary supplementation with grape residue flour on weight gain, metabolic profile, leukogram, proteinogram and antioxidant response in suckling lambs.

The objective of this study was to determine whether the addition of grape residue flour (GRF) in the diet of suckling lambs would improve their health and consequently enhance their growth. We used 48 lambs, 30 days of age divided into four treatments with four repetitions each, with each repetition consisting of three animals. The groups were identified as follows: G-0%, used as a control (without GRF) and G-0.5%, G-1% and G-2% referring to treatment with grape residue flour in doses of 0.5%, 1%, and 2% of inclusion in the concentrate, respectively. Lambs in groups G-1% and G-2% had higher final body weights, weight gain and average daily gain (ADG) compared to the other groups. GRF dietary supplementation had good antioxidant potential, being able to stimulate glutathione S-transferase (GST) activity and consequently reduce levels of reactive oxygen species (ROS) in lambs that consumed the highest dose of GRF (G-2%). GRF supplementation improved humoral responses, with increased serum levels of heavy-chain and light-chain immunoglobulins; however, there was a reduction in serum ceruloplasmin levels in these lambs. We observed higher concentrations of glucose and triglycerides in lambs in the 2% group. There were lower lymphocyte counts in lambs that received GRF. Lambs supplemented with the highest doses of GRF (G-1% and G-2%) had lower total bacterial counts in the feces. We conclude that the use of GRF in the supplementation of suckling lambs improved animal health, as it stimulated the antioxidant and immune systems and consequently favored their growth.

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.

Experimental infection with Escherichia coli in broilers: Impacts of the disease and benefits of preventive consumption of a stimulator of homeopathic immunity.

Colibacillosis is a disease caused by Escherichia coli that manifests itself when there are homeostatic imbalances or in the context of increased exposure, in which case the organism displays opportunistic behavior. To control this problem in poultry, antibiotics are used in the feed, because E. coli is component of the intestinal microbiota of birds. However, because of the changing dietary habits of the human population that seeks out healthier foods without antimicrobial residues, there have been many studies of alternatives to replace conventional antimicrobials as performance enhancers. Thus, the objective of the present study was to determine whether daily consumption of a homeopathic product (immune stimulator) by broilers stimulates immune responses and thereby minimizes the negative effects of experimental E. coli infection. We used 320 1-day-old Cobb 500 chicks, distributed in two groups with eight repetitions each, and 20 birds per repetition: control (CG) and homeopathy (HG). HG birds consumed doses of 0.02 mL/bird/day (1-7 d) via water, 0.01 ml/bird (8-21 d), 0.02 ml/bird (22-28 d), 0.01 mL/bird (29-35 d), and 0.02 mL/bird (35-45 d), as recommended by the manufacturer. At day 22 of the birds' life, the two groups were divided into four subgroups, with four repetitions per subgroup. On day 22, birds in CG1 and HG1 groups were infected intraperitoneally with 0.5 mL of inoculum containing 1.0 × 108 CFU of E. coli/mL. During the experimental period, data were collected for analysis of performance. On days 21 and 45 of age, we collected blood and feces. During the first 21 days of the experiment, we found that birds that consumed the immunostimulator had lower neutrophil counts and higher levels of globulins, however without significant difference between groups in terms of performance. Uninfected birds that consumed the homeopathic product in the water had less feed conversion (HG2) between days 1-35 and 1 to 45 compared to the other treatments. Mortality was higher in groups experimentally infected with E. coli (HG1 and CG1) from 22 to 35 days of life. There were greater numbers of lymphocytes in the HG2 group on day 45 than in CG1 and CG2; while numbers of neutrophils were lower at 42 days in birds of groups HG1 and HG2 than in CG1. Lower total bacterial counts, total coliforms and E. coli were observed in the feces of birds in the HG2 group compared to the other groups. Taken together, these findings suggest that inclusion of homeopathic product in the water of broilers had positive effects on the modulation of the immune response and on feed conversion in birds not challenged with E. coli. But the preventive protocol used in this study was not able to minimize the negative effects caused by the experimental E. coli intraperitoneal infection in broilers, featuring a substantial infectious challenge.

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.

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.

Daily consumption of a homeopathic product decreases intestinal damage and stool bacterial counts in mice challenged with Escherichia coli.

Escherichia coli is a bacterium normally found in the gastrointestinal tract of domestic animals that can usually control the infection. Nevertheless, some factors (high exposure, stress conditions, animal category, among others) can favor the exacerbation of E. coli infection and cause of disease. Because it is a zoonotic bacterium, it is important to control the infection, avoiding contamination of home interiors in the case of pets. There are various forms of treatment for E. coli; nevertheless, there are few options for prevention. In the present study, we evaluated homeopathy. Thus, the objective of this study was to determine whether administration of a prophylactic homeopathic in water would minimize the negative effects of E. coli infection, as well as reducing bacterial counts in the feces of a experimental model. Forty mice were divided into four experimental groups (n = 10/group). Groups NC (negative control) and PC (positive control) were not treated; in group T1, the animals received 0.002 mL/day/animal of the homeopathic in water, and animals in group T2 0.004 mL/day/animal. The experiment lasted 54 days, and on the 31st day, mice of T1, T2 and PC groups were infected orally a 0.2 mL inoculum of 1.5 × 108 CFU of E. coli. Euthanasia and sample collection were performed on the 40th and 54th days of the experiment (n = 5/group/time point). Blood, liver, spleen, intestine, and feces samples were collected from the final portion of the intestine. There was no significant difference in animal weight between groups at the end of the experiment. Neutrophil count was lower in PC group animals on day 40, while on day 54, the counts were lower in T2 and PC. Lymphocyte counts were lower only in the PC group than in the NC group on day 54. Globulins were lower in the NC and PC groups than in T1 and T2 on day 40, remaining lower the PC group and higher in T1 on day 54; levels of immunoglobulin IgG and IgM were higher in groups T1 and T2, which differed from PC and NC. TNF-α levels were higher in the T1 and T2 groups at 40 and 54 days. INF-γ levels were higher in T1, T2, and PC compared to NC on day 40, remaining higher than NC in groups T1 and T2 on day 54. Total bacterial count, total coliforms and E. coli counts were lower in group T1 and higher in NC and PC on days 40 and 54, when they were lower for T1 and T2. Histologically, no lesions were observed in extra-intestinal tissues; however the height of intestinal crypts in the PC group was smaller than the others on day 40. On day 54, villi and crypts of all infected groups were larger in T1 and T2 than in NC; sizes in the PC group were higher than those of all other groups. These data suggest that the homeopathic agent in the drinking water improved health of the mice.

Calidad del calostro bovino y su relación con la genética, el manejo, la fisiología y su congelación / Quality of bovine colostrum and its relation to genetics, management, physiology and its freezing

RESUMEN Objetivo. Evaluar si la calidad del calostro se modifica por las características genéticas, fisiológicas y de manejo en el período preparto, así como evaluar si la calidad y composición del calostro se alteran en el proceso de congelación. Material y métodos. En el experimento I, se recogieron muestras de calostro y sangre de 35 vacas (18 Holstein y 17 Jersey). En el experimento II, se recolectaron seis muestras de calostro de vacas Holstein y se congelaron durante 60 días. Resultados. La concentración media de inmunoglobulina G (IgG) fue de 77.65 mg/ml en Jersey y de 82.77 mg/ml en Holstein. La genética, el orden de parto y la interacción entre estos factores no fueron significativos en la concentración de IgG en el calostro. Además, se observó un efecto genético de la vaca en el peso en la cría al nacer y en los tres días de edad (p<0.0001). Con respecto a la transmisión de inmunidad pasiva de terneros, no se observaron efectos de la raza de la vaca y el orden de parto en la concentración de proteínas plasmáticas de la ternera, así como después de tres días de congelación. Las razas de becerros Holstein (83%) y Jersey (82%) mostraron niveles de proteína sérica total por encima de 5.5 g/dL. Las vacas Holstein alojadas en potreros individuales con suplementos dietéticos, proporcionaron una mejor calidad de calostro (93.57 mg IgG/mL). Con el tiempo, el porcentaje de grasa cambió al congelarse, que se redujo con el tiempo (p<0.05) en el Experimento II. Conclusiones. El manejo previo al parto influye en la calidad del calostro, y la congelación no interfiere en la calidad centesimal e inmunológica del calostro, con excepción de la grasa, que disminuye a lo largo del tiempo.

ABSTRACT Objective. The aims of this study were to assess whether colostrum quality is modified by genetic, physiological and management characteristics in the pre-partum period, as well as evaluate whether quality and composition of colostrum is altered in the freezing process. Material and methods. In the experiment I, colostrum and blood samples of 35 cows (18 Holstein and 17 Jerseys) were collected. In the experiment II, six colostrum samples of Holstein cows were collected and frozen during 60 days. Results. The mean immunoglobulin (Ig) concentration was 77.65 mg/ml to Jersey and 82.77 mg/ml to Holstein. The genetic, parturition order, and the interaction between these factors were no significant on IgG concentration in the colostrum. Also, it was observed an effect genetic of cow in the weight on calf at birth and on three days of age (p<0.0001). Regarding transmission of calf passive immunity, no effects of cow breed and calving order were observed on plasma protein concentration of calf, as well as after three days of freezing. Calves of Holstein (83%) and Jersey (82%) breed showed total serum protein levels above 5.5 g/dL. Holstein cows housed in individual paddocks with diet supplementation provided better quality of colostrum (93.57 mg Ig/mL). Over time, the percentage of fat reduced at freezing, that reduced over time (p<0.05) in Experiment II. Conclusions. The pre-partum management exerts influence on colostrum quality, and the freezing not interfere on centesimal and immunological quality of colostrum, with exception the fat, that decrease along the time.

Dietary addition of curcumin favors weight gain and has antioxidant, anti-inflammatory and anticoccidial action in dairy calves / La adición de curcumina en la dieta de los terneros lecheros favorece el aumento de peso y tiene acción antioxidante, antiinflamatoria y anticoccidial / A adição de curcumina na dieta dos bezerros leiteiros favorece o aumento de peso e tem ação antioxidante, anti-inflamatória e anticoccidiostática

Abstract Background: Curcumin has been used as an additive in the diet of animals in recent years due to the potent medicinal properties of this molecule. Objective: To evaluate whether the addition of curcumin to the diet of calves at different phases (pre- and post-weaning) has a positive effect on metabolic profile, performance, and anti-coccidian action. Methods: Thirty-three Holstein calves were selected at various phases of development: Experiment 1 (E1: n=10) 18±7 (pre-weaning), Experiment 2 (E2: n=11) 64±4 (pre-weaning) and Experiment 3 (E3: n=12) 95±8 (post-weaning) days of life. The calves were separated in three groups according to their phase of development. In each experiment, animals were divided into two sub-groups: control and curcumin. The curcumin groups received 200 mg of additive per animal/day either in milk (pre-weaning) or concentrate (post-weaning). Fecal collections were performed on days 0, 10 and 15 of the experiment to count Eimeria oocysts per gram of feces and to perform fecal score analysis. Complete blood counts, oxidant and antioxidant profiles, protein metabolism markers, lipid levels, glucose levels, and animal weights were measured. Analyses of digestibility and composition of the diet used in Experiment 3 (post-weaning) were also performed. Results: Independent of phase, animals that received curcumin had greater weight gain on days 0 to 15 (E1, E2 and E3 p=0.04, 0.001 and 0.001, respectively), probably due to the increased digestibility of hay and concentrate at 72h (p=0.03 and 0.02, respectively). The supplemented calves had lower level of oxidants (thiobarbituric acid reactive substances -TBARS- and reactive oxygen species -ROS-), indicating that free radical levels in serum and lipid peroxidation were lower. This was probably due to increased enzymatic antioxidants gluthatione S-transferase (E1, E2 and E3 p=0.001, 0.001 and 0.02, respectively), catalase (E1 p=0.001) and superoxide dismutase (E3 p=0.001) in treated animals at day 15. Furthermore, calves receiving curcumin had lower numeric number of Eimeria infection during the experimental period, and the difference was significant in day 15 (E1 and E2 p=0.02, and 0.001, respectively). Conclusion: Curcumin supplementation to dairy calves has coccidiostatic potential, favoring weight gain.

Resumen Antecedentes: Curcumina ha sido utilizada como aditivo en la dieta de animales en los últimos años, debido a las potentes propiedades medicinales de esa molécula. Objetivo: Evaluar si la adición de curcumina en la alimentación de terneras en diferentes fases (pre y post-destete) presenta efecto positivo sobre el perfil metabólico, desempeño de los animales, y acción anti-coccidial. Métodos: Se seleccionaron 33 terneros Holstein en varias etapas de desarrollo: Experimento 1 (E1: n=10) 18±7 (pre-destete), Experimento 2 (E2: n=11) 64±4 (pre-destete) y Experimento 3 (E3: n=12) 95±8 (post-destete) días de vida. Para todos los experimentos, el período experimental fue de 15 días. Los animales se dividieron en dos grupos: control y tratados con curcumina. Los grupos con curcumina recibieron una dosis de 200 mg del aditivo por animal/día en la leche (pre-destete) o en el concentrado (post-destete). Las colectas de heces y sangre fueron realizadas en los días 0, 10 y 15 para conteo de ooquistes de Eimeria por gramo de heces y análisis de puntaje fecal. Se realizó hemograma, perfil oxidante y antioxidante, metabolismo proteico, lipídico, glucosa sanguínea y pesaje de los animales. También se realizó análisis de digestibilidad de la dieta total para los animales del Experimento 3 (post-destete). Resultados: Encontramos que, independientemente de la fase, los animales que recibieron curcumina tuvieron una mayor ganancia de peso en los días 0 a 15 (E1, E2 y E3, p=0,04, 0,001 y 0,001, respectivamente), probablemente debido al aumento de la digestibilidad del heno y concentrado a las 72 h (p=0,03 y 0,02, respectivamente). En los terneros suplementados observamos un nivel más bajo de oxidantes (oxidantes (sustancias reactivas al ácido tiobarbitúrico y especies reactivas de oxígeno); es decir, los niveles séricos de radicales libres y la peroxidación lipídica fueron más bajos. Esto se debió, probablemente, a los antioxidantes enzimáticos glutatión S-transferasa (E1, E2 y E3 p=0,001, 0,001 y 0,02 respectivamente), catalasa (E1, p=0,001) y superóxido dismutasa (E3, p=0,001) que aumentaron en los animales tratados al día 15. Además, los terneros que recibieron curcumina tuvieron niveles más bajos de infección por Eimeria durante el período experimental y fueron significativos en el día 15 (E1 y E2, p=0,02 y 0,001, respectivamente). Conclusión: La suplementación con curcumina tiene potencial coccidiostático y favorece la ganancia de peso en terneros Holstein.

Resumo Antecedentes: Curcumina tem sido usado como aditivo na dieta de animais nos últimos anos, devido as propriedades medicinais potente dessa molécula. Objetivo: Avaliar se a adição de curcumina na alimentação de bezerras em diferentes fases (pré e pós-desmame) apresenta efeito positivo sobre perfil metabólico, desempenho e ação anti-coccidéo. Métodos: Para isso, 33 bezerros holandeses foram selecionados em vários estágios de desenvolvimento: Experimento 1 (E1: n=10) 18±7 (pré-desmame), Experimento 2 (E2: n=11) 64±4 (pré-desmame) e Experimento 3 (E3: n=12) 95±8 (pós-desmame) dias de vida. Para todos os experimentos o período experimental foi de 15 dias, assim como foram delineados com dois grupos: controle e tratados com curcumina. Os grupos de curcumina receberam 200 mg do aditivo por animal/dia no leite (pré-desmame) ou em concentrado (pós-desmame). Coletas de fezes e sangue foram realizadas nos dias 0, 10 e 15 de experimento para contagem de oocistos de Eimeria por grama de fezes e análise de escore fecal. Do sangue colhido foram realizados: hemograma, perfil oxidante e antioxidante, metabolismo proteico, lipídico, glicose, além da pesagem dos animais. Também foi realizado análise de digestibilidade da dieta total ofertada aos animais do Experimento 3 (pós-desmame). Resultados: Independentemente da fase, os animais que receberam curcumina tiveram maior ganho de peso do dia 0 a 15 (E1, E2 e E3, p=0,04, 0,001 e 0,001, respectivamente), provavelmente devido ao aumento da digestibilidade ao feno e concentrado após 72 h (p=0,03 e 0,02, respectivamente). Nos bezerros suplementados, observou-se menor nível de oxidantes (substâncias reativas ao ácido tiobarbitúrico e espécies reativas de oxigênio), ou seja, os níveis séricos de radicais livres e a peroxidação lipídica foram menores. Isto foi provavelmente devido a antioxidantes enzimáticos glutationa S-transferase (E1, E2 e E3, p=0,001, 0,001 e 0,02, respectivamente), catalase (E1, p=0,001) e superóxido dismutase (E3, p=0,001) aumentando nestes animais tratados no dia 15. Além disso, bezerros recebendo curcumina tiveram menores níveis de infecção por Eimeria durante o período experimental e significativos no dia 15 (E1 e E2, p=0,02 e 0,001, respectivamente). Conclusão: A suplementação de curcumina aumenta o potencial coccidiostático e favorece o ganho de peso.

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.

Dietary vegetable choline improves hepatic health of Nile tilapia (Oreochromis niloticus) fed aflatoxin-contaminated diet.

Aflatoxin B1 (AFB1) is one of the most important mycotoxins due to its hepatotoxic and carcinogenic effects on animals. The effect of dietary supplementation with vegetable choline (VC) at 400, 800, and 1200 mg/kg against the deleterious effects of AFB1 (2 ppm/kg diet) in the liver of Nile tilapia (Oreochromis niloticus) was studied. The experimental period was 81 days, and the diet with VC was offered to the fish for 60 days prior to challenge with AFB1. Diets with AFB1 were tested in three replications and animals were analyzed at days 14 and 21 of dietary intake. The addition of VC to tilapia diet increased body weight (days 30 and 60 pre-challenge and day 21 post-challenge). The group fed aflatoxin-contaminated diet presented significantly reduced antioxidant enzymes and increased reactive oxygen species (ROS) levels, thiobarbituric acid reactive species (TBARS) levels, and protein carbonyl (PC) content in the liver. Dietary supplementation with VC at 800 and 1200 mg/kg demonstrated a significant protective effect, avoiding the increase of ROS, TBARS, and PC levels in the liver of tilapia from the aflatoxin contaminated groups. Thus, dietary VC supplementation may be used in tilapia to increase antioxidant status and reduce the negative effects caused by AFB1 toxicity. Based on the findings, it is recommended to use VC as a food supplement for Nile tilapia in order to avoid AFB1 toxication. In addition, decreased aflatoxin toxicity can be attributed to the VC antioxidant property.

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.

Impacts of Escherichia coli infection in young breeder chicks on the animal behavior and cerebral activity of purinergic and cholinergic enzymes involved in the regulation of molecules with neurotransmitter and neuromodulator function.

The objective of this study was to evaluate if infection by Escherichia coli in juvenile breeder chicks alters the activity of enzymes involved in neurotransmission and cerebral immunomodulation, including acetylcholinesterase (AChE), nucleoside triphosphate diphosphohydrolase (NTPDase), 5'-nucleotidase (5'NT) and adenosine deaminase (ADA), as well as their effects on the pathogenesis of the disease. We divided 20 growing breeder chicks into two groups (n = 10 per group). One group was experimentally infected with 1 mL of culture medium containing 1 × 108 CFU of E. coli intraperitoneally. The other was the negative control. On the tenth day after infection, the animals were euthanized and brain samples were collected. Macroscopically, pericarditis and hepatic congestion were observed in the birds, but without histopathological lesions in the encephalon although the bacterium was present in the cerebral cortex of all animals in the infected group (i.e., they were PCR-positive). The activity of AChE, NTPDase, 5'-NT and ADA were evaluated in the cerebral homogenates of the birds after 10 days of infection. AChE activity in the cerebral cortex was lower in the infected group than in the control; there was an increase in the activity of NTPDase, 5'-nucleotidase and ADA, possibly indicating greater hydrolysis of ATP (P < 0.001), ADP (P < 0.01) and AMP (P < 0.01), followed by increased adenosine deamination (P < 0.001). Despite these changes, no apparently diseased animals were observed throughout the experimental period. Therefore, such changes in enzymatic activity may affect the functioning of the central nervous system because these enzymes are responsible for extracellular regulation of molecules that act on neurotransmission and immunomodulation such as acetylcholine, ATP and adenosine.

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.