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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.

Curcumin-containing nanocapsule powder formulations have not been used in ruminant feed to date, despite the fact that curcumin is known to be a functional food additive. The objective of this study was to determine whether ethyl polymethacrylate (Eudragit L-100) nanocapsules loaded with curcumin (N-CU) would improve health and growth of lambs. Thirty-two male Lacaune lambs (body weight [BW] = 16 ± 0.99 kg; 45 d of age) were randomly assigned to 1 of 4 treatments: T0, T1, T2 and T4, representing supplementation of curcumin at 0, 1, 2, and 4 mg/kg concentrate, respectively. The animals in each treatment were allocated in 4 pens of 2 lambs each (8 lambs per treatment). The experiment lasted 17 d, with samples and measurements collected on d 0, 7, 12, and 17. The T2 lambs had greater average daily gain than T0 lambs. Regression analysis showed that the ideal dose of N-CU to enhance weight gain was 1.89 mg/kg concentrate. There were significant interactions (P

This study evaluated the effects of curcumin dietary supplementation on growth, physiology, carcass characteristics and meat quality in lambs. Thirty-two male Lacaune lambs (15.6 ± 0.63 kg, 60 ± 2.8 days of age) were randomly allocated in 16 pens (four treatments of four replicates with two lambs each) for 30 days. The animals were assigned to the following treatments: T0, T100, T200 and T300, representing 0, 100, 200 and 300 mg of curcumin/kg of concentrate, respectively. Curcumin dietary supplementation increased (P = 0.02) weight gain; on regression analysis, 315.1 mg curcumin/kg of concentrate was the dosage that provided the greatest weight gain. T200 and T300 lambs had lower (P = 0.04) erythrocytes numbers; T100 and T300 lambs had lower (P = 0.01) leukocyte numbers and T300 lambs had lower (P = 0.04) lymphocyte numbers, compared to T0 lambs. Globulin levels were significantly greater in the T200 group (P = 0.04) only on day 15 but not day 30; levels of total protein were significantly higher (P = 0.01) only in T200 and T300 group on day 15 and only in T200 group on d 30. Gamma-glutamyltransferase concentrations tended to be lower (P = 0.08) in T100, T200 and T300 group on d 15, and only in the T100 group on d 30. Curcumin dietary supplementation increased (P = 0.01) the serum activity of antioxidant enzymes and reduced (P = 0.01) levels of reactive oxygen species. In meat samples, T200 and T300 had greater total antioxidant capacity (P = 0.03) and lower (P = 0.01) lipoperoxidation rates. In carcasses, T300 lambs had greater (P ≤ 0.02) cooling weight losses and yields than did T0 lambs. Curcumin dietary supplementation also reduced (P ≤ 0.03) redness and yellowness. T200 and T300 lambs had fewer (P = 0.01) cooking losses and T200 lambs had greater (P = 0.03) water holding capacity than did T0 lambs. These findings suggest that curcumin dietary supplementation improves growth and antioxidant responses, as well as influencing meat quality in lambs.

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.

The objective of this study was to determine whether snacks containing curcumin have beneficial effects on dog health. The snacks were produced from commercial canned meat for dogs, and curcumin was added and homogenized. Snacks containing 15 mg of curcumin were produced, frozen, and offered to dogs twice a day. Ten beagles (6 months of age) were used. The animals were placed in an experimental kennel and allocated randomly to 1 of 2 treatments (5 dogs/treatment): snacks containing curcumin (curcumin; 30 mg of curcumin/animal/day) or not (control). On day 15, numbers of erythrocytes and hematocrit were greater in dogs fed with curcumin than in control dogs. Dogs fed with curcumin had lower numbers of leukocytes (day 30), neutrophils (day 15), and lymphocytes (day 30) than did control dogs. Dogs fed with curcumin had lower plasma levels of nitric oxide, reactive oxygen species, lipoperoxidation, and protein carbonylation on day 30 than did control dogs. Finally, dogs fed with curcumin had greater plasma total antioxidant capacity and concentrations of protein thiol, non-protein thiol, glutathione peroxidase, and superoxide dismutase on day 30 compared with control dogs. We conclude that curcumin in dog snacks stimulated the antioxidant system and consequently reduced oxidative reactions, which is beneficial to animal health. Furthermore, 30 mg of curcumin/dog/day reduced leukocyte counts, which suggests mild anti-inflammatory effects.

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.

Bee honey has beneficial properties that are recognized worldwide, including high nutritional value. The objective of this study was to determine whether a daily dose of honey would improve productive performance as well as energy, protein, and lipid metabolisms in lactating lambs. Bee honey (from Apis mellifera) was offered orally for the first 7 days of the animals’ lives. Twenty Lacaune lambs were used, allocated to two groups: group C, lambs not supplemented with honey, and group T, consisting of lambs fed daily doses of 10 mL of honey. The experimental period lasted 45 days. Body weight measurements and blood sample collections were performed at five time points (days 1, 7, 15, 30, and 45 of life). The lambs that consumed honey showed significantly greater weight gain during the experimental period; there was greater weight gain between days of life 30 and 45. We found significantly higher levels of urea (days 30 and 45) and globulins (day 45). Carbohydrate (glucose), lipid (triglycerides and cholesterol), and protein (total protein and albumin) levels did not differ between treatments. Based on these preliminary results, we conclude that supplementation with honey in the lamb diets may stimulate growth in during lactating phase.

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.

The pathophysiological mechanisms associated with immune responses to the opportunistic bacterial pathogen Providencia rettgeri in fish species are unknown. The aim of this study was to evaluate whether purinergic signaling generates anti-inflammatory or pro-inflammatory profiles in lymphoid immune organs of the Nile tilapia (Oreochromis niloticus) experimentally infected by P. rettgeri. Adenosine triphosphate (ATP) and adenosine diphosphate (ADP) hydrolysis activities in the spleen and triphosphate diphosphohydrolase (NTPDase) in the head kidney decreased in experimentally P. rettgeri-infected Nile tilapia on days seven and fourteen post-infection (PI) compared to the control group. At these same time points, splenic and head kidney adenosine deaminase (ADA) activity, and levels of ATP, interleukin-15 (IL-15) and metabolites of nitric oxide (NOx) increased in infected animals compared to their respective control groups; this was also observed for serum ATP levels. No significant difference was observed between groups regarding 5′-nucleotidase activity. These results suggest that purinergic signaling in lymphoid immune organs of Nile tilapia exerts a pro-inflammatory profile in response to P. rettgeri infection. This response contributes to the inhibition of immune and inflammatory responses by reduction of ATP hydrolysis and augmentation of ATP in the extracellular environment as well as augmentation in adenosine deamination and its possible reduction in the extracellular environment, leading to a self-sustained pro-inflammatory cycle, corroborated by excessive content of ATP (extracellular milieu), IL-15 and NOx levels. Purinergic signaling may thereby contribute to the pathophysiology of P. rettgeri infection.

The objective of this study was to determine whether the addition of acai (Euterpe oleracea) oil in the diets of lactating sheep under heat stress exerted beneficial effects on health as well as milk production and quality. Eighteen multiparous Lacaune sheep (2 or 3 parities; 28–30 days of lactation; average milk production of 1.7 L/sheep/day) were stratified by parity and milk production and were assigned randomly to 1 of 2 treatments (9 sheep/treatment): diet supplemented with 2% of soybean oil (SOY) or 2% of acai oil (ACAI) in the concentrate for 14 days. The amount of oil added in the diet was equivalent to 0.65% of the total diet (dry matter basis). Blood and milk samples were collected on days 1, 10 and 14. On day 14, the ACAI group sheep had lower serum concentrations of leukocytes, neutrophils, and lymphocytes than did the SOY group sheep. On day 14, ACAI group sheep had lower serum concentration of triglycerides and urea, milk concentration of fat and total solid and milk lipid peroxidation than did SOY group sheep. However, on day 14, ACAI group sheep had higher serum concentrations of glucose and globulin, serum and milk antioxidant capacity against peroxyl radicals, milk production and productive efficiency than did SOY group sheep. The fatty acids profile in milk did not differ between groups. These data suggest that acai oil improved the antioxidant activity in serum and milk and improved milk production and quality in dairy sheep under heat stress.

The objective of this study was to evaluate whether the antiparasitic drug eprinomectin causes hepatic oxidative stress and impairment of hepatic energetic metabolism in the silver catfish (Rhamdia quelen), in conditions of experimental aquatic contamination. The fish were exposed for 24 and 48 h at various concentrations of eprinomectin (0.0, 1.124, 1.809 and 3.976 μg L−1), followed by 48 h of recovery in eprinomectin-free water. We measured levels of reactive oxygen species (ROS) and lipoperoxidation (LOOH) in hepatic tissue, as well as levels of antioxidant capacity against peroxyl radicals (ACAP), and activities of the antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione S-transferase (GST), in addition to the activities of enzymes involved in energy metabolism, adenylate kinase (AK) and pyruvate kinase (PK). After 24 h, hepatic ROS levels increased in fish exposed to all concentrations of eprinomectin, and the same occurred with LOOH at the two highest concentrations of eprinomectin. ACAP levels decreased after 24 h of exposure to the highest concentration, also occurring with SOD activity at the two highest concentrations. Glutathione enzymes activities (GPx and GST) decreased after 48 h of exposure to the highest concentration. Fish transfer to eprinomectin-free water, with re-analysis after 48 h post-recovery, was unable to reverse ROS and LOOH increases, likewise with the reductions of ACAP, SOD, GPx and GST, especially at the two highest concentrations. AK and PK activities decreased after 48 h of exposure, AK decreased at the two highest concentrations and PK decreased at the highest concentration. AK activity remained lower after 48 h in eprinomectin-free water. PK activity returned to normal after 48 h in eprinomectin-free water. In summary, silver catfish exposed to aquatic contamination with the antiparasitic drug eprinomectin showed hepatic oxidative stress, possibly related to impairment of the phosphotransfer network and energy metabolism.

Bacterial diseases are one of the major problems in freshwater fish culture and have been linked to significant losses and high mortality rate. In this study, Nile tilapia Oreochromis niloticus was infected by Providencia rettgeri to evaluate the oxidative stress and antioxidant responses in the fish tissues. Juvenile Nile tilapia was divided into two groups, as follow: control (uninfected) and experimentally infected with 100 μL of P. rettgeri suspension containing 2.4 × 107 viable cells/fish, and the liver and kidney tissues were collected on days 7 and 14 post-infection (PI). Liver and kidney ROS and lipid peroxidation levels were high in infected fish on day 14 PI compared to control group, while superoxide dismutase activity was lower in liver (days 7 and 14 PI) and kidney (day 14 PI) compared to their respective control groups. Liver and kidney antioxidant capacity against peroxyl radicals, non-proteic, and proteic thiols levels was lower in infected tilapia on day 14 PI compared to control group. Based on these results, P. rettgeri infection may elicit oxidative damage via increased ROS production, decreased ROS elimination and inhibits enzymatic and non-enzymatic antioxidant defense systems; which may contribute directly to disease pathophysiology of infected animals.

The objective of this study was to evaluate whether antiparasitic eprinomectin may be an environmental contaminant in water compartment in low concentrations, negatively affecting neurotransmission and, consequently, the natural behavior of the jundia (Rhamdia quelen). Fish were randomly allocated in tanks and exposed for 24 and 48 h to eprinomectin concentrations in water [0.0 (Control), 1.124 (T1), 1.809 (T2) and 3.976 (T3) μg L−1], followed by 48 h of recovery in eprinomectin-free water, in order to evaluate the behavioral parameters, levels of reactive oxygen species (ROS) in the brain, as well as cerebral enzymatic activities of acetylcholinesterase (AChE) and of the sodium-potassium ATPase pump (Na+/K+-ATPase). Especially at the two highest concentrations of eprinomectin (T2 and T3), the fish showed alterations in natural behavior, particularly hyperlocomotion and longer time on the surface. Furthermore, at these same concentrations, cerebral ROS levels increased and cerebral AChE activity decreased. At the highest concentration (T3) cerebral Na+/K+-ATPase activity was reduced. Increased ROS and impairment of AChE and Na+/K+-ATPase enzymes in the brain may have contributed directly to behavioral changes, due to neuronal damage and synapse impairment. Even after 48 h in water without eprinomectin, behavioral changes and neurotoxic effects were observed in fish, suggesting residual effects of the antiparasitic. In conclusion, eprinomectin even in low concentrations may be a hazardous environmental contaminant for aquatic organisms, as it causes brain damage and affects the natural behavior of fish.

Grape pomace is a by-product of the winemaking process, rich in bioactive compounds such as resveratrol and has antioxidant and antimicrobial properties. Recent evidence has demonstrated the protective effects of grape against impairment of the phosphoryl transfer network, a system linked to energetic homeostasis of high-energy phosphoryl transfer, to maintain energetic balance. Also, Pseudomonas aeruginosa infection in fish has been related to severe impairment of the branchial phosphoryl transfer network via inhibition of creatine kinase (CK), adenylate kinase (AK) and pyruvate kinase (PK) activities. Thus, the aim of this study was to evaluate whether dietary supplementation with grape pomace flour (GPF) is able to reduce or prevent the impairment of cellular energetic homeostasis in grass carp (Ctenopharyngodon idella) experimentally infected by P. aeruginosa. Branchial CK (cytosolic and mitochondrial), AK, and PK activities were lower in infected animals on day 15 post-infection (PI) compared to uninfected fish, while lactate dehydrogenase (LDH) activity was higher. Branchial reactive oxygen species (ROS), thiobarbituric acid reactive substances (TBARS), lipid oxidation (LOOH) and protein carbonylation levels were higher, while the level of non-proteic (NPSH) and proteic (PSH) thiols was lower in infected fish on day 15 PI, as compared to uninfected fish. The use of 300 mg of GPF/kg of feed was able to ameliorate the activities of CK, AK, PK, and LDH as compared to infected fish, but their activities remained different from the control group. On the other hand, dietary supplementation with 300 mg of GPF/kg of feed was able to prevent all alterations linked to oxidative damage elicited by disease. Based on this evidence, inhibition of the phosphoryl transfer network may contribute to the impairment of intracellular energetic communication between cellular ATP synthesis and consumption and is mediated by oxidative damage in the gills of fish infected by P. aeruginosa. The use of 300 mg of GPF/kg of feed exerted protective effects on branchial energetic metabolism linked to ATP metabolism by reducing the impairment in cellular energetic homeostasis; its effects can be mediated by the prevention of SH group oxidation.

The objective of this study was to evaluate the effects of grape residue flour (GRF) on antioxidant activities, biochemistry variables, components of the immune system and milk production and quality of Lacaune sheep in heat stress. Twenty-seven multiparous lactating sheep [50 ± 1.8 days (d) milking] were stratified by initial body weight, age, date of lambing and milk production and assigned randomly to 1 of 3 treatments (9 sheep/treatment): no GRF supplementation (control group) or supplementation at 1% (10 g/kg GRF) or 2% (20 g/kg GRF) of GRF (bark and seed) in the concentrate (grains and minerals mixture). Each ewe received 0.8 kg/d of concentrate, 3.6 kg/d of corn silage, and 0.25 kg/d of Cynodon spp hay. Milk production along with blood and milk samples were collected on d 1, 10 and 15. The 2% GRF sheep had increased serum concentrations of superoxide dismutase and glutathione peroxidase activity on d 15 compared to control sheep. Over time (d 10 to 15), lipid peroxidation was reduced in 2% GRF sheep. Total serum antioxidant capacity was greater in 2% GRF sheep compared to control sheep on d 10 and 15. Superoxide dismutase and glutathione peroxidase activity in milk samples were greater in 2% GRF sheep compared to control sheep. Supplementation with GRF did not affect milk production but GRF sheep were more efficient compared to control sheep. Protein and lactose concentrations were similar between treatments, but total solids and fat concentrations were greater in 2% GRF sheep compared to control sheep on d 15. Somatic cell count was reduced in GRF sheep compared to control sheep. In summary, supplementation with 2% GRF in dairy sheep in heat stress resulted in antioxidant and anti-inflammatory responses, which improved milk quality and reduced somatic cell count and lipid peroxidation.

Fish known as Curimbata (Prochilodus lineatus) suffered a reduction in natural stocks due to predatory fishing and environmental deterioration; these are among the reasons for increased interest in the inclusion of this species in agricultural fairs. The aim of this study was to evaluate whether transport and exposure to an agricultural fair alters plasma biochemical parameters in curimbatas. Blood samples were collected at 0 h (before transport), 10 h (end of transport), and 72 h (end of exposure to agricultural fair). Plasma cortisol levels increased after 72-h exposure to agricultural fair compared with initial collection (0 h). Plasma reactive oxygen species levels were lower in fish at 72-h exposure to agricultural fairs compared with those at initial collection, while plasma thiobarbituric acid reactive substances levels were higher after 10 h of transport than at initial collection (0 h). Plasma superoxide dismutase and glutathione S-transferase activities were lower after 10 h of transport than at initial collection, while plasma glutathione peroxidase activity was higher in fish exposed for 72 h at the agricultural fair compared with initial collection (0 h). Plasma total protein, albumin, and glucose levels were higher after 10 h of transport than at initial collection, while triglycerides levels were higher after 10 h of transport and 72-h exposure to the agricultural fair than at initial collection. The transport of these fish (average weight 209 g) for display agricultural fairs causes stress demonstrated by biochemical changes and increases in antioxidant enzyme activity, resulting in decreases of reactive oxygen species.

The aim of this study was to evaluate whether rupture on blood-brain barrier (BBB) can be a pathway for trichlorfon-induced neurotoxic effects, and to investigate its implications on oxidative status, cell viability and brain neurotransmitters in silver catfish (Rhamdia quelen). The BBB permeability was increased in fish exposed for 24 h to 22 mg/L of trichlorfon compared to the control group, as well as in those exposed to 11 and 22 mg/L of trichlorfon for 48 h. Compared to the control group, brain reactive oxygen species and lipid peroxide levels were higher when exposed to 22 mg/L of trichlorfon and 11 and 22 mg/L of trichlorfon after 24 h and 48 h, respectively, while the antioxidant capacity against peroxyl radical levels was lower. Exposure to 22 mg/L of trichlorfon for 24 h reduced brain cell viability compared to the control group, together with 11 and 22 mg/L of trichlorfon for 48 h. Also, brain AChE, Na+ and K+-ATPase activities were reduced in those fish exposed to trichlorfon compared to the control group. Thus, the rupture of BBB can be considered an important pathway involved in trichlorfon-induced neurotoxic effects, which contributes to brain oxidative damage and important changes on brain neurotransmitters.

Pseudomonas aeruginosa is a Gram-negative opportunistic bacterial pathogen in aquaculture systems being associated to extensive liver damage caused by oxidative stress in both marine and freshwater fish. Dietary supplementation with natural antioxidants is considered a rational strategy to prevent hepatic diseases involved with oxidative stress. Bio-residues resulting from the wine industry, such as grape pomace, are potential sources of bioactive phenolic compounds that can be applied as supplement for animal production. Thus, the aim of this study was to evaluate whether dietary supplementation with grape pomace flour (GPF) was able to prevent or reduce the hepatic oxidative damage of grass carp, Ctenopharyngodon idella, experimentally infected by P. aeruginosa. Hepatic reactive oxygen species (ROS), metabolites of nitric oxide (NOx), thiobarbituric acid reactive substances, and protein carbonylation levels were higher in fish experimentally infected by P. aeruginosa compared to the control group. Hepatic superoxide dismutase and catalase activities and antioxidant capacity against peroxyl radical levels were also higher in fish experimentally infected by P. aeruginosa compared to the control group. Dietary supplementation with 300 mg/kg GPF prevented all alterations elicited by P. aeruginosa, with the exception of protein carbonylation levels. The dietary supplementation with 150 mg/kg GPF was not able to avoid alteration of the analyzed variables, being results similar to those infected (positive control). Based on these results, dietary supplementation with 300 mg/kg GPF prevented P. aeruginosa-induced liver damage in grass carp, and this protective effect occurred through prevention on excessive ROS and NOx production, as well as via prevention of lipid damage. Moreover, 300 mg/kg GPF exerted its hepatoprotective effects by improving enzymatic and non-enzymatic antioxidant defense system. In summary, this supplementation can be an interesting approach to prevent P. aeruginosa-induced liver damage.

It is becoming evident that bacterial infectious diseases negatively affect gill energy metabolism, mainly because it causes alterations of enzymes that belong to the phosphotransfer network, such as creatine kinase (CK), adenylate kinase (AK) and pyruvate kinase (PK). Thus, the aim of this study was to evaluate whether the phosphotransfer network could be one pathway involved in the bioenergetic imbalance during Providencia rettgeri infection, as well as to investigate the bacteria's influence on adenosine triphosphate (ATP)-dependent enzymes. In this study, branchial cytosolic and mitochondrial CK activity decreased in infected animals on day 14 post-infection (PI) when compared to its respective control group. Moreover, branchial sodium-potassium pump (Na+/K+-ATPase) and proton-pump (H+-ATPase) activities decreased in infected animals on day 14 PI compared to the control. Finally, branchial thiobarbituric acid reactive substance levels increased in infected animals on day 14 PI, while non-protein thiol (NPSH) levels decreased. No significant difference was observed between groups regarding AK and PK activities. In summary, the impairment of CK activity elicited by P. rettgeri caused an impairment in branchial energetic homeostasis, reducing ATP availability in the gills. Moreover, inhibition of CK activity provokes an impairment of ATP-dependent enzymes, such as Na+/K+-ATPase and H+-ATPase, which can be mediated by lipid oxidative damage and reduction of NPSH content.

Hypoxia is a critical issue in aquaculture of marine or freshwater fishes especially in intensive aquaculture systems, contributing to severe economic losses for fish producers. Some evidence has suggested the involvement of phosphoryl transfer network, catalyzed by creatine kinase (CK), pyruvate kinase (PK) and adenylate kinase (AK) in the metabolic alterations during exposure to hypoxia, but the effects on fish exposed to acute hypoxia remains unknown. Moreover, some evidence has revealed the protective effects of caffeine supplementation against hypoxia, including linked to energetic status. Therefore, the aim of this study was to evaluate whether diet supplemented with caffeine (5 and 8%) is able to protect the gills against bioenergetics dysfunction on Nile tilapia exposed to hypoxia. Branchial cytosolic and mitochondrial CK, and sodium-potassium pump (Na+, K+-ATPase) activities were inhibited in fish exposed to hypoxia compared to control group, while branchial AK and lactate dehydrogenase (LDH) activities and lactate levels were stimulated. The use of 8% dietary caffeine was able to prevent all alterations induced by exposure to hypoxia, with exception of the inhibition on Na+, K+-ATPase activity. The lower activity to both CK fractions (cytosolic and mitochondrial) contributes to impairment on bioenergetic homeostasis, while augmentation on AK activity can be considered an attempt to prevent or reduce the energetic imbalance during hypoxia. Also, the use of 8% dietary caffeine prevented alteration energetic metabolism, as well as occurrence of anaerobic metabolism. In summary, use of 8% dietary caffeine can be an alternative to improve branchial energetic homeostasis during exposure to hypoxia.

Mancozeb is a fungicide widely used in agriculture, mostly against the pathogen Glomerella cingulata responsible for the rot of ripe grape, but presents high toxicity. Strategies are sought to reduce the toxicity of this fungicide and alternative treatments are welcome. An alternative could be the use of clove oil, which has Eugenol as its major compound, and has antifungal potential against G. cingulata, however, Eugenol is susceptible to degradation processes which may compromise its efficacy. The nanoencapsulation of Mancozeb and Eugenol is a possible strategy to overcome the limitations of toxicity, solubility and instability of these compounds. Therefore, the objective of this study is to develop nanoemulsions containing Mancozeb (0.1 mg/mL) and Eugenol (33 mg/mL), isolated or associated, and evaluate the safety of these formulations through cytotoxicity, genotoxicity and ecotoxicity tests. Nanoemulsions were developed by the spontaneous emulsification method, cytotoxicity and genotoxicity were evaluated in healthy human cells through MTT, Dichlorofluorescein diacetate and Picogreen tests, and ecotoxicity assessment was carried out using the chronic toxicity test in springtails. After preparation, the physicochemical characterization of the nanoemulsions were performed which presented mean particle size between 200 and 300 nm, polydispersity index less than 0.3, negative zeta potential and acid pH. The nanoencapsulation was able to avoid the reduction of the cell viability caused by Mancozeb, while Eugenol was shown to be safe for cell use in both free and nanostructured forms, however the association of the two active compounds showed toxicity in the higher doses of Mancozeb. In the ecotoxicity tests, both free Mancozeb and Eugenol forms presented high toxic potential for soil, whereas the nanoencapsulation of these compounds did not cause a reduction in number of springtails. Therefore, from the tests performed, it was possible to observe that nanoencapsulation of Mancozeb and Eugenol is a safe alternative for the application of these compounds mainly in agriculture.

The spleen is a secondary lymphatic organ that has an important role in fish immune and inflammatory responses. Reports have suggested a role of purinergic signaling in immune and inflammatory responses during bacterial infection; however, the role of this pathway during Pseudomonas aeruginosa infection remains unknown. The aim of this study was to investigate the involvement of purinergic signaling in the pathogenesis of P. aeruginosa infection of grass carp, Ctenopharyngodon idella. In addition, grape pomace flour (GPF), a bio-residue of the wine industry, was assessed as a dietary supplement and whether it was able to modulate purinergic signaling and improve the immune and inflammatory responses of C. idella, experimentally infected with P. aeruginosa. Nucleoside Triphosphate diphosphohydrolase (NTPDase) activity was downregulated in the serum and spleen of fish experimentally infected with P. aeruginosa, compared to the control group. By contrast, serum and splenic adenosine deaminase (ADA) activity was stimulated in fish experimentally infected with P. aeruginosa compared to the control group. Dietary supplementation with GPF was able to prevent all alterations induced by infection. Serum and splenic metabolites of nitric oxide (NOx) were higher in fish experimentally infected with P. aeruginosa compared to the control group, and GPF supplementation was able to prevent this alteration. Serum and splenic myeloperoxidase and catalase activities were also higher in fish experimentally infected with P. aeruginosa compared to the control group, and again, the use of GPF was able to prevent these alterations. Based on these metabolic changes, purinergic signaling contributed to the pro-inflammatory profile of immune lymphatic organs during P. aeruginosa infection and to disease pathophysiology. Dietary supplementation with GPF modulated the inflammatory response by altering serum and splenic purinergic signaling. GPF may be means of improving immune and inflammatory responses resulting from P. aeruginosa infection of grass carp.

This study aimed to produce dog food containing natural antioxidants (blend of essential oils and vitamin E) to replace synthetic antioxidants and determine the effects on food conservation and animal health sequentially. The foods were produced in a commercial factory, and the antioxidants were added at the oil bath stage. Ten adult beagle dogs were used, divided into two treatments; control treatment (CON; synthetic antioxidant feed [butylhydroxytoluene]) and test treatment (NAT; natural antioxidant feed; blend of essential oils from clove, rosemary, oregano, and vitamin E). The dogs were weighed at the beginning and end of each experimental period, and there were no treatment effects for body weight. In both treatments, food conservation efficiency was observed, demonstrating the feasibility of using natural sources as antioxidants in dog food because chemical and oxidative variables did not differ regardless of the antioxidant used during production. The animals’ metabolic and haematological variables were not influenced by the treatments; however, a reduction in the number of lymphocytes was observed over time only in the dogs of the NAT group. There was also a day effect for total leukocyte, neutrophil and erythrocyte counts only in NAT animals, which means a significant increase (p ≤ 0.05) in the variables on d 28. There was an effect of the treatment and an interaction treatment x day for the total bacterial count, whereas a decrease in the bacterial count (p p ≤ 0.05) to minimise oxidative stress. In group NAT, the NPSH and glutathione S-transferase levels were increased, which may explain the decrease in ROS levels. It was concluded that natural antioxidants in dog feed, in addition to promoting feed conservation, stimulate levels of systemic antioxidants and minimise the impacts caused by free radicals in the dogs’ blood.

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.

The objective of this work was to determine whether preventive consumption of a homeopathic product via drinking water would protect mouse kidneys from a challenge with the nephrotoxic antibiotic gentamicin. We used 40 Swiss mice divided into four groups with ten animals each. The homeopathic product was supplied through water for 30 days in a preventive manner and gentamicin for 10 days to induce an experimental renal failure. The groups were as follows: Negative-CT (homeopathic and gentamicin was not provided), Positive-CT (did not receive homeopathic, but received gentamicin 40 mg/kg), T2 (received 0.002 ml of the product per animal/day, and received gentamicin 40 mg/kg), and T4 (0.004 ml of the product per animal/day and received gentamicin 40 mg/kg). On days 12 and 20, blood and tissue samples were collected from five animals in each group. No histopathological lesions were found in mouse kidneys. However, levels of thiobarbituric acid reactive substances, reactive oxygen species, and nitrite/nitrate ratios in the kidney of the Positive-CT group were higher compared to the other groups. As for glutathione S-transferase, on the 20th day, the groups that used the homeopathic product (T2 and T4) had higher activities than the positive TC. Therefore, the results suggest that prophylactic consumption of the hepatoprotective homeopathic product can decrease lipid peroxidation, nitrous stress, and oxidative stress at the renal level when consecutive doses of gentamicin induce insufficiency. El objetivo de este trabajo fue determinar si el consumo preventivo de un producto homeopático a través del agua potable protegería los riñones de los ratones de un desafío con el antibiótico nefrotóxico gentamicina. Usamos 40 ratones suizos divididos en cuatro grupos con diez animales cada uno. El producto homeopático se suministró a través de agua durante 30 días de forma preventiva y gentamicina durante 10 días para inducir una insuficiencia renal experimental. Los grupos fueron los siguientes: CT negativo (no se proporcionó homeopático y gentamicina), CT positivo (no recibió homeopático, pero recibió gentamicina 40 mg/kg), T2 (recibió 0,002 ml del producto por animal/día, y recibió gentamicina 40 mg/kg) y T4 (0,004 ml del producto por animal / día y recibió gentamicina 40 mg/kg). Los días 12 y 20, se recogieron muestras de sangre y tejido de cinco animales de cada grupo. No se encontraron lesiones histopatológicas en riñones de ratón. Sin embargo, los niveles de sustancias reactivas con ácido tiobarbitúrico, especies reactivas de oxígeno y relaciones nitrito/nitrato en el riñón del grupo de CT positiva fueron más altos en comparación con los otros grupos. En cuanto a la glutatión S-transferasa, el día 20, los grupos que utilizaron el producto homeopático (T2 y T4) tuvieron actividades más altas que los TC positivos. Por lo tanto, los resultados sugieren que el consumo profiláctico del producto homeopático hepatoprotector puede disminuir la peroxidación lipídica, el estrés nitroso y el estrés oxidativo a nivel renal cuando dosis consecutivas de gentamicina inducen insuficiencia. O objetivo deste trabalho foi determinar se o consumo preventivo de um produto homeopático via água potável protegeria os rins de camundongos de um desafio com o antibiótico nefrotóxico gentamicina. Foram utilizados 40 camundongos Swiss divididos em quatro grupos com dez animais cada. O produto homeopático foi fornecido por meio de água por 30 dias de forma preventiva e gentamicina por 10 dias para induzir insuficiência renal experimental. Os grupos foram os seguintes: TC-negativa (não foi fornecida gentamicina e homeopática), TC-positiva (não recebeu homeopatia, mas recebeu gentamicina 40 mg/kg), T2 (recebeu 0,002 ml do produto por animal/dia, e receberam gentamicina 40 mg/kg) e T4 (0,004 ml do produto por animal/dia e receberam gentamicina 40 mg/kg). Nos dias 12 e 20, amostras de sangue e tecido foram coletadas de cinco animais em cada grupo. Nenhuma lesão histopatológica foi encontrada em rins de camundongos. No entanto, os níveis de substâncias reativas ao ácido tiobarbitúrico, espécies reativas de oxigênio e relações nitrito/nitrato no rim do grupo CT-positivo foram maiores em comparação com os outros grupos. Já para a glutationa S-transferase, no 20º dia, os grupos que usaram o produto homeopático (T2 e T4) apresentaram atividades superiores ao TC positivo. Portanto, os resultados sugerem que o consumo profilático do produto homeopático hepatoprotetor pode diminuir a peroxidação lipídica, o estresse nitroso e o estresse oxidativo no nível renal quando doses consecutivas de gentamicina induzem insuficiência.

We determined whether the dietary supplementation with a commercial product (vegetable biocholine - VB) for Lacaune ewes at peak lactation would improve animal health and production as well as milk quality. We also determined the effects of VB as a feed additive. We used thirty Lacaune ewes at 30 days of lactation, allocated into three groups: T0 (control, without VB), T5 (5 g of VB/animal/day) and T10 (10 g of VB/animal/day). T10 sheep had greater milk yield during the experimental period. T10 ewes had also a significantly lower feed conversion ratio than the other groups. Total solids concentration of milk was higher in T10 than in the other groups on day 20. The somatic cell count in milk was lower in ewes that consumed VB in the highest dose (T10) than in the T0 and T5 in days 15 and 20. Lower levels of lipoperoxidation (LPO) and of reactive oxygen species (ROS) were observed in milk from ewes fed with the VB supplemented diet on day 20, associated with increased levels of total antioxidant capacity and superoxide dismutase activity in milk. In serum, we found that T5 and T10 animals had significantly higher levels of non-enzymatic antioxidants (non-protein thiols) associated with reduced LPO and ROS content. Higher levels of globulins were observed in T10 than in T0 sheep on day 20. These data suggest that VB dietary supplementation stimulates antioxidant responses and increases the concentration of globulins in a manner beneficial to sheep health. Milk production, feed conversion rate, and milk SCC were also improved., Antioxidant; Production; Lactation; Health animal; Agricultural biotechnology.

We evaluated performance, nutrient balance, nitrogen balance, and serum parameters in pigs in the nursery phase raised in environmental conditions above the thermoneutral zone that were fed with diets containing various levels of crude protein (CP). A total of 15 barrows (22.75 ± 1.58 kg) were housed in metabolism pens and distributed in a completely randomized design with three treatments: 15.5%, 18.3%, and 21.0% of CP, designated T15, T18, and T21, respectively. There was a gradual increase of temperature over the first three days from 25 to 29.1 ± 2.3 °C. From d18 to d20 of the experiment, pigs received the same diet (18% CP) and thermoneutral conditions were reestablished (22.9 ± 1.9 ºC). There were higher values of urinary, excreted, and absorbed nitrogen in T21, followed by T18 and T15. Blood urea levels were higher in treatments with higher protein levels. On d4 (adaptation period), cholesterol levels were higher in the T15 group than in the T21 group, antioxidant power of iron reduction values were lower in the T18 group than the T21 group. Advanced oxidation protein products (AOPP) on day 16 were higher in the T15 group, associated with the accumulation of heat stress and lower CP diets. Similar results were obtained for T18 with higher AOPP values on d16 than on d8 and d12. However, there were greater AOPP values in the T21 group on d20 (when the CP level was reduced to 18%) than on d8. Amino acid supplementation and reduction of CP in the diet to levels of 15.5% in piglets raised above the thermoneutral zone improved the use of CP. Furthermore, 15.5% of CP in the diet reduced the excretion of urinary nitrogen. The N-retention was not affected by dietary CP level, using the ideal protein concept. The use of 21%-CP in the diet efficiently avoided exacerbation of protein oxidation for pigs raised above thermoneutrality. Evaluamos el desempeño, balance de nutrientes, balance de nitrógeno y parámetros séricos en cerdos en fase de crianza criados en condiciones ambientales por encima de la zona termoneutra que fueron alimentados con dietas conteniendo varios niveles de proteína cruda (PC). Un total de 15 carretillas (22,75 ± 1,58 kg) se alojaron en corrales de metabolismo y se distribuyeron en un diseño completamente al azar con tres tratamientos: 15,5%, 18,3% y 21,0% de PC, designados T15, T18 y T21, respectivamente. Hubo un aumento gradual de la temperatura durante los primeros tres días de 25 a 29,1 ± 2,3 °C. Desde el d18 al d20 del experimento, los cerdos recibieron la misma dieta (18% PC) y se restablecieron las condiciones termoneutras (22,9 ± 1,9 ºC). Hubo valores más altos de nitrógeno urinario, excretado y absorbido en T21, seguido de T18 y T15. Los niveles de urea en sangre fueron más altos en tratamientos con niveles más altos de proteínas. En el d4 (período de adaptación), los niveles de colesterol fueron más altos en el grupo T15 que en el grupo T21, el poder antioxidante de los valores de reducción de hierro fueron más bajos en el grupo T18 que en el grupo T21. Los productos de proteína de oxidación avanzada (AOPP) el día 16 fueron más altos en el grupo T15, asociados con la acumulación de estrés por calor y dietas más bajas de PC. Se obtuvieron resultados similares para T18 con valores de AOPP más altos en d16 que en d8 y d12. Sin embargo, hubo mayores valores de AOPP en el grupo T21 el día 20 (cuando el nivel de PC se redujo al 18%) que el día 8. La suplementación con aminoácidos y la reducción de PC en la dieta a niveles del 15,5% en lechones criados por encima de la zona termoneutra mejoraron el uso de PC. Además, el 15,5% de la PC en la dieta redujo la excreción de nitrógeno urinario. La retención de N no se vio afectada por el nivel de PC en la dieta, utilizando el concepto de proteína ideal. El uso de 21% de PC en la dieta evitó eficazmente la exacerbación de la oxidación de proteínas en cerdos criados por encima de la termoneutralidad. Avaliamos o desempenho, o balanço de nutrientes, o balanço de nitrogênio e os parâmetros séricos em suínos na fase de creche criados em condições ambientais acima da zona termoneutra, alimentados com dietas contendo diferentes níveis de proteína bruta (PB). Um total de 15 suínos machos castrados (22,75 ± 1,58 kg) foram alojados em baias de metabolismo e distribuídos em um delineamento inteiramente casualizado com três tratamentos: 15,5%, 18,3% e 21,0% de PB, designados T15, T18 e T21, respectivamente. Houve um aumento gradual da temperatura nos primeiros três dias de 25 para 29,1 ± 2,3 °C. Do d18 ao d20 do experimento, os suínos receberam a mesma dieta (18% PB) e as condições termoneutras foram restabelecidas (22,9 ± 1,9 ºC). Houve maiores valores de nitrogênio urinário, excretado e absorvido em T21, seguido de T18 e T15. Os níveis de uréia no sangue foram maiores nos tratamentos com níveis mais elevados de proteína. No d4 (período de adaptação), os níveis de colesterol foram maiores no grupo T15 do que no grupo T21, o poder antioxidante do valor de redução do ferro foi menor no grupo T18 do que no grupo T21. Os produtos proteicos de oxidação avançada (AOPP) no dia 16 foram maiores no grupo T15, associados ao acúmulo de estresse térmico e dietas com menor PB. Resultados semelhantes foram obtidos para T18 com valores de AOPP mais altos em d16 do que em d8 e d12. No entanto, houve maiores valores de AOPP no grupo T21 no d20 (quando o nível de PB foi reduzido para 18%) do que no d8. A suplementação de aminoácidos e a redução da PB na dieta para níveis de 15,5% em leitões criados acima da zona termoneutra melhoraram o uso de PB. Além disso, 15,5% de PB na dieta reduziu a excreção de nitrogênio urinário. A retenção de N não foi afetada pelo nível de PB da dieta, usando o conceito de proteína ideal. O uso de 21%-PB na dieta evitou de forma eficiente a exacerbação da oxidação de proteínas em suínos criados acima da termoneutralidade.

The objective of this study was to determine whether a phytogenic blend (PB), formulated based on organic acids, tannins, curcumin, and essential oils, could replace the antimicrobials commonly used as growth promoters in the poultry industry without compromising zootechnical performance, health, or meat quality. In addition, our goal was to report the anti-aflatoxin effect of this phytogenic blend. Four treatments were used: TC, or control; T250, T500, and T1000, representing test doses of 250, 500, 1000 mg PB/kg of feed, respectively, or a 34-day experiment (initial and growth phases). On day 22 of the study and age of the birds, 500 ppb of aflatoxin was included in the diet to represent an intestinal challenge and to evaluate the growth-promoting effects of PB. In the initial phase (up to 21 days), there were no differences between groups in weight gain, feed intake, or feed conversion. After adding an aflatoxin-contaminated feed, doses of 250 and 500 mg/kg minimized the adverse effects on feed consumption and feed conversion caused by aflatoxin; but 1000 mg/kg did not differ between groups. In birds that consumed PB (T250, T500, and T1000) compared to the control, there were the following changes: 1) lower counts of heterophiles, lymphocytes, and monocytes; 2) lower lipid peroxidation and high non-protein thiols levels in breast meat; 3) lower bacteria counts in broiler litter; and 4) lower ALT levels. Greater intestinal villus/crypt ratios were observed at T250 and T500. The dose of 250 mg/kg reduced saturated fatty acids and increased unsaturated fatty acids. The chemical-physical composition of the meat did not differ between treatments. The findings suggest that the addition of a PB has a high potential to improve performance for chickens in the growing stage and minimize the adverse effects of aflatoxicosis.

The aim of this study was to determine whether infection by Clostridium perfringens negatively interferes with the oxidant/antioxidant status and activity of energy metabolism enzymes (creatine kinase (CK), adenylate kinase (AK) and pyruvate kinase (PK)), as well as the zootechnical performance of broilers. A completely randomized design with three treatments, with five replications by treatment, and 10 birds by repetitions was used: T1: non-infected group; T2: group infected with C. perfringens; T3: group infected with C. perfringens, and basal diet with performance enhancers (antibiotic and coccidiostatic). At 21 days of age, the birds were experimentally infected orally with 4.0 x 108 CFU/mL C. perfringens. At strategic moments during the experimental period, zootechnical data and blood samples were collected. There were no significant differences among treatments in terms of weight gain, feed intake, or feed conversion. At 20 days, heterophils counts were significantly lower in the T3 group birds than in the others. A significant reduction in the number of total leukocytes, because of the lower number of lymphocytes, was observed in groups T2 and T3 on day 34. Neutrophil and monocyte counts were significantly lower in group T3 than in group T1 on day 34. On day 42, there were significantly lower levels of total protein and globulin in birds in T3 than in T1. On day 27, PK activity was significantly higher in groups T2 and T3; similar behavior observed on day 34 when CK and PK activities in T2 and T3 were significantly higher than those of T1. AK activity was significantly higher in the serum of T3 birds than in those of the other groups at 34 days. At 41 days, CK and PK activities were significantly higher only in birds in group T3 compared to other groups. The levels of reactive oxygen species and lipoperoxidation, as well as the activity of glutathione S-transferase, were significantly lower in birds from group T3 compared to other groups at age 20 days. After a challenge with C. perfringens, there were fluctuations in the behavior of oxidative stress biomarkers; in particular, at 41 days of age, birds in T2 had elevated levels of lipoperoxidation, different from what was seen in birds in T3 that consumed feed with a performance enhancer. Preliminary results suggest that clostridiosis affects serum activity of enzymes in the phosphotransfer network, requiring compensatory enzymatic changes in an attempt to maintain energy homeostasis. In addition, the infection reduces the number of inflammatory cells and causes oxidant/antioxidant imbalance that may contribute to the pathophysiology of the disease. Además, la infección reduce el número de células inflamatorias y provoca un desequilibrio en el estado oxidante / antioxidante que puede contribuir a la fisiopatología de la enfermedad. El objetivo de este estudio fue determinar si la infección por Clostridium perfringens interfiere negativamente con el estado oxidante / antioxidante y la actividad de las enzimas del metabolismo energético (creatina quinasa (CK), adenilato quinasa (AK) y piruvato quinasa (PK)), así como comportamiento zootécnico de pollos de engorde. O delineamento experimental foi inteiramente casualizado com três tratamentos, com cinco repetições por tratamento e 10 pollos por repetições: T1: grupo no infectado; T2: grupo infectado por C. perfringens; T3: grupo infectado por C. perfringens y dieta basal con promotores del crecimiento (antibiótico y coccidiostático). A los 21 días de edad, las aves fueron infectadas experimentalmente por vía oral con 4.0 x 108 UFC/mL de C. perfringens. En momentos estratégicos del período experimental, fueron recolectaron datos zootécnicos y muestras de sangre. No hubo diferencias significativas entre los tratamientos en términos de aumento de peso, consumo de alimento y conversión alimenticia. A los 20 días, los recuentos de heterófilos fueron significativamente más bajos en las pollos del grupo T3 que en las otras pollos. Se observó una reducción significativa en el número de leucocitos totales, debido al menor número de linfocitos, en los grupos T2 y T3 el día 34. Los recuentos de neutrófilos y monocitos fueron significativamente más bajos en el grupo T3 que en el grupo T1 el día 34. El día 42, hubo niveles significativamente más bajos de proteína total y globulina en pollos de T3 en comparación con T1. El día 27, la actividad PK fue significativamente mayor en los grupos T2 y T3; comportamiento similar observado en el día 34 cuando las actividades de CK y PK en T2 y T3 fueron significativamente mayores en relación con T1. La actividad de AK fue significativamente mayor en el suero de pollos de engorde T3 en comparación con los otros grupos a los 34 días. A los 41 días, las actividades de CK y PK fueron significativamente más altas sólo en las aves del grupo T3 en comparación con los otros grupos. Los niveles de especies reactivas al oxígeno y la lipoperoxidación, así como la actividad de la glutatión S-transferasa, fueron significativamente menores en los pollos del grupo T3 en comparación con otros grupos a los 20 días de edad. Después de un desafío con C. perfringens, hubo fluctuaciones en el comportamiento de los biomarcadores de estrés oxidativo; en particular, a los 41 días de edad, los pollos de T2 mostraron altos niveles de lipoperoxidación, diferente a lo observado en pollos de T3 que consumieron alimento con promotores de crecimiento. Estos resultados preliminares sugieren que la clostridiosis afecta la actividad sérica de las enzimas en la red de fosfotransferencia, lo que requiere cambios enzimáticos compensatorios en un intento por mantener la homeostasis energética. O objetivo deste estudo foi determinar se a infecção por Clostridium perfringens interfere negativamente no status oxidante / antioxidante e na atividade das enzimas do metabolismo energético (creatina quinase (CK), adenilato quinase (AK) e piruvato quinase (PK)), bem como no desempenho zootécnico de frangos de corte. O delineamento experimental foi inteiramente casualizado com três tratamentos, com cinco repetições por tratamento e 10 aves por repetições: T1: grupo não infectado; T2: grupo infectado com C. perfringens; T3: grupo infectado com C. perfringens e dieta basal com promotores de crescimento (antibiótico e coccidiostático). Aos 21 dias de idade, as aves foram infectadas experimentalmente por via oral com 4,0 x 108 UFC/mL de C. perfringens. Em momentos estratégicos do período experimental, foram coletados dados zootécnicos e amostras de sangue. Não houve diferenças significativas entre os tratamentos em termos de ganho de peso, consumo de ração e conversão alimentar. Aos 20 dias, as contagens de heterófilos foram significativamente menores nas aves do grupo T3 do que nas outras. Foi observada uma redução significativa no número de leucócitos totais, devido ao menor número de linfócitos nos grupos T2 e T3 no dia 34. As contagens de neutrófilos e monócitos foram significativamente menores no grupo T3 do que no grupo T1 no dia 34. No dia 42, houve níveis significativamente menores de proteína total e globulina nas aves do T3 em relação ao T1. No dia 27, a atividade da PK foi significativamente maior nos grupos T2 e T3; comportamento semelhante observado no dia 34 quando as atividades de CK e PK em T2 e T3 foram significativamente maiores em relação ao T1. A atividade de AK foi significativamente maior no soro das aves do T3 em comparação aos outros grupos aos 34 dias. Aos 41 dias, as atividades de CK e PK foram significativamente maiores apenas nas aves do grupo T3 em comparação com os outros grupos. Os níveis de espécies reativas ao oxigênio e lipoperoxidação, bem como a atividade da glutationa S-transferase, foram significativamente menores nas aves do grupo T3 em relação com os outros grupos aos 20 dias de idade. Após um desafio com C. perfringens, houve flutuações no comportamento dos biomarcadores de estresse oxidativo; em particular, aos 41 dias de idade, as aves do T2 apresentaram níveis elevados de lipoperoxidação, diferente do que foi observado em aves do T3 que consumiram ração com promotores de crescimento. Estes resultados preliminares sugerem que a clostridiose afeta a atividade sérica de enzimas da rede de fosfotransferência, o que exige alterações enzimáticas compensatórias na tentativa de manter a homeostase energética. Além disso, a infecção reduz o número de células inflamatórias e causa desequilíbrio no status oxidante/antioxidante que pode contribuir para a fisiopatologia da doença.

The aim of this study was to determine whether the addition of a homeopathic product to dairy cow feed would improve health and production efficiency, as well as composition and quality of milk. The product tested here was produced to stimulate immunity and indirectly to prevent the colonization of the mammary gland by bacteria, thereby decreasing somatic cell counts and bacteria in cow’s milk. Fifty dairy cows were used, divided into two groups: Control (n = 25) and Treated (n = 25). For 90 consecutive days, 50 grams of homeopathic product (treated group) was added to the concentrate; 50 g of the product vehicle (limestone) was added to the concentrate of the control cows. We performed composition and quality analysis on the milk (days 1, 15, 30, 45, 60, 75, and 90) as well as blood collection to carry out hematological and metabolism analyses (days 1, 30, 60, and 90). We found that fat content in the milk cows consuming the homeopathic agent was greater than that of the control group. Because of this increased fat content, there was a tendency towards higher total solids content in treated cows. A treatment effect was found in terms of total bacterial count (TBC); that is, there were lower TBCs in milk of treated animals than in control animals. There was a trend of a treatment effect versus day for somatic cell count (SCC); that is, there were lower SCC in animals that consumed the homeopathic on days 15 and 30 of the experiment. There was no difference between groups in terms of milk production; however, there was a negative correlation between milk production and TBC or SCC in the milk of cows that consumed homeopathic product. Animals in the treated group had lower total leukocyte and lymphocyte counts than did the control group, as well as a tendency toward lower neutrophil counts in these animals. Several bacteria were isolated from the cows' milk during the experimental period, with no treatment effect. In particular, we isolated Corynebacterium spp., Staphylococcus aureus, Staphylococcus epidermidis and Staphylococcus hyicus in greater numbers of cows compared to other etiologic agents. Based on these results, we conclude that consumption of the homeopathic product by dairy cows had positive effects on milk quality. El objetivo de este estudio fue determinar si la adición de un producto homeopático a la alimentación de las vacas lecheras mejoraría la salud y la eficiencia de producción, así como la composición y calidad de la leche. El producto probado aquí fue producido para estimular la inmunidad e indirectamente para prevenir la colonización de la glándula mamaria por bacterias, disminuyendo así el recuento de células somáticas y bacterias en la leche de vaca. Se utilizaron cincuenta vacas lecheras, divididas en dos grupos: Control (n = 25) y Tratadas (n = 25). Durante 90 días consecutivos, se añadieron al concentrado 50 gramos de producto homeopático (grupo tratado); Se añadieron 50 g del vehículo producto (piedra caliza) al concentrado de las vacas de control. Realizamos análisis de composición y calidad de la leche (días 1, 15, 30, 45, 60, 75 y 90) así como extracción de sangre para realizar análisis hematológicos y metabólicos (días 1, 30, 60 y 90). Encontramos que el contenido de grasa en las vacas lecheras que consumían el agente homeopático era mayor que el del grupo de control. Debido a este mayor contenido de grasa, hubo una tendencia hacia un mayor contenido de sólidos totales en las vacas tratadas. Se encontró un efecto del tratamiento en términos de recuento bacteriano total (RBT); es decir, hubo RBT más bajos en la leche de los animales tratados que en los animales de control. Hubo una tendencia de un efecto del tratamiento frente al día para el recuento de células somáticas (RCS); es decir, hubo menor RCS en los animales que consumieron el homeopático los días 15 y 30 del experimento. No hubo diferencia entre los grupos en términos de producción de leche; sin embargo, hubo una correlación negativa entre la producción de leche y RBT o RCS en la leche de las vacas que consumieron productos homeopáticos. Los animales del grupo tratado tenían recuentos de leucocitos y linfocitos totales más bajos que el grupo de control, así como una tendencia hacia recuentos de neutrófilos más bajos en estos animales. Se aislaron varias bacterias de la leche de vaca durante el período experimental, sin efecto del tratamiento. En particular, aislamos Corynebacterium spp., Staphylococcus aureus, Staphylococcus epidermidis y Staphylococcus hyicus en un mayor número de vacas en comparación con otros agentes etiológicos. Con base en estos resultados, concluimos que el consumo del producto homeopático por las vacas lecheras tuvo efectos positivos sobre la calidad de la leche. O objetivo deste estudo foi determinar se a adição de um produto homeopático à ração para vacas leiteiras melhoraria a saúde e a eficiência da produção, bem como a composição e qualidade do leite. O produto testado aqui foi produzido para estimular a imunidade e indiretamente para prevenir a colonização da glândula mamária por bactérias, diminuindo assim a contagem de células somáticas e bactérias no leite de vaca. Foram utilizadas 50 vacas leiteiras, divididas em dois grupos: Controle (n = 25) e Tratada (n = 25). Por 90 dias consecutivos, 50 gramas do produto homeopático (grupo tratado) foram adicionados ao concentrado; 50 g do veículo produto (calcário) foram adicionados ao concentrado das vacas controle. Foram realizadas análises de composição e qualidade do leite (dias 1, 15, 30, 45, 60, 75 e 90), bem como coleta de sangue para realização de análises hematológicas e metabólicas (dias 1, 30, 60 e 90). Descobrimos que o teor de gordura nas vacas leiteiras que consumiam o agente homeopático era maior do que no grupo controle. Devido ao aumento do teor de gordura, houve uma tendência de maior teor de sólidos totais nas vacas tratadas. Um efeito do tratamento foi encontrado em termos de contagem bacteriana total (CBT); ou seja, houve menor CBT no leite dos animais tratados do que nos animais controle. Houve uma tendência de efeito do tratamento versus dia para contagem de células somáticas (CCS); ou seja, houve menor CCS nos animais que consumiram o homeopático nos dias 15 e 30 do experimento. Não houve diferença entre os grupos em termos de produção de leite; entretanto, houve uma correlação negativa entre a produção de leite e CBT ou CCS no leite de vacas que consumiram produtos homeopáticos. Os animais do grupo tratado apresentaram menor contagem total de leucócitos e linfócitos do que o grupo controle, bem como tendência para menor contagem de neutrófilos nesses animais. Várias bactérias foram isoladas do leite de vaca durante o período experimental, sem efeito do tratamento. Em particular, isolamos Corynebacterium spp., Staphylococcus aureus, Staphylococcus epidermidis e Staphylococcus hyicus em maior número de vacas em comparação com outros agentes etiológicos. Com base nesses resultados, concluímos que o consumo do produto homeopático por vacas leiteiras teve efeitos positivos na qualidade do leite.

Made available in DSpace on 2020-12-12T01:24:53Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-10-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa e Inovação do Estado de Santa Catarina Brazil is the number one exporter of chicken meat, and this industry maintains constant microbiological vigilance. The objective of this study was to characterize the pathogenicity, antimicrobial resistance (AMR) and the profile of biofilm production of Escherchia coli strains isolated from raw refrigerated cuts of chicken meat sold in retail markets of the four largest poultry companies in Brazil. We collected 150 samples of chicken meat, in order to isolate E. coli and performed susceptibility tests (to amoxicillin associated with clavulanic acid, ceftiofur, enrofloxacin, gentamicin, and trimethoprim + sulfamethoxazole). In addition, the disc approximation test to detect extended spectrum beta-lactamases enzymes (ESBLs) producers was performed. E. coli ability to form biofilm was checked using polystyrene microplates. We also searched for ESBLs genes (blaCTY-M2, blaSHV-1, blaTEM-1, blaCTX-M2, blaOXA-1, blaPSE-1 and AmpC) and adhesion genes (sfa/foc, afa/draB, iha, hrla, fimC, tsh, papC, mat, cr1, felA, fimH and papG) in ESBL-E. coli producers and in those E. coli classified as strongly biofilm formers, respectively. The overall percentage of E. coli isolation was 58.66%, with brand A having the highest percentage (70%), followed by brands D, B and C (60, 53.3 and 50%, respectively). The highest resistance profile was observed for beta-lactams (39.5%), followed by sulfonamide associated to trimethoprim (36.9%) and polymyxin (33.4%). Of the isolates obtained, 77% were non-susceptible to at least one antimicrobial. Brand A showed the highest overall percentage of resistance with 95.23%, followed by brands C (80%), B (75%) and D (69.44%). Overall, 73.86% of the isolates were non susceptible to at least one antibiotic and 36.3% were multiresistants. A total of 17.04% of E. coli strains were identified as ESBLs producers and 70.44% were able to form biofilms (moderate-to-strong). The blaTEM-1 gene was the most prevalent (73.33%), followed by blaSHV-1 (46.66%) and blaCMY-2 (6%). Of the 31 strongly biofilm-forming strains, 26 (83.87%), 24 (77.41%) and 20 (64.51%) expressed fimC, papG and crl genes, respectively. Taken together, our results show that Brazilian chicken meat can be contaminated with E. coli that are non-susceptible to multiple antibiotics, able to form biofilm and showing a diverse repertoire of adhesins linked to pathogenicity depending on the brand evaluated. Graduate Program in Animal Science Universidade do Estado de Santa Catarina (UDESC-Oeste) Universidade Federal do Rio Grande do Sul (UFRGS) Laboratory of Applied Statistics Department of Mathematics São Paulo State University (Unesp) School of Engineering Laboratory of Applied Statistics Department of Mathematics São Paulo State University (Unesp) School of Engineering

This study aimed to assess the antimicrobial activity of various honeys against strains of gram-negative and -positive bacteria, as well as to determine the physicochemical parameters of these honeys. Seven honeys from various species of stingless bees were evaluated. The physical-chemical parameters evaluated were pH, moisture, water activity, acidity, ash, electrical conductivity and color. Antimicrobial activity was determined using disc diffusion agar tests and minimum inhibitory concentrations. We found that there was a relationship between the physical-chemical parameters and the antimicrobial activity. The minimum inhibitory concentration of 25% honey was able to inhibit the growth of both gram-positive and -negative bacteria; the greatest efficacy was verified for the species of bees Melipona mondury, M. quadrifasciata, Scaptotrigona bipunctata and Tetragona clavipes. Regarding synergism, Escherichia coli maintained its sensitivity profile in relation to all studied honeys combined with antimicrobials. An important factor to consider is the concentration of honey capable of sensitizing the microorganism, as it has been shown to be dependent on the species of the stingless bee. Nevertheless, all honeys showed antimicrobial activity in various methods of analysis. These data suggest that honey is a promising alternative to sensitize resistant microorganisms, for the health of humans and animals alike. Este estudio tuvo como objetivo evaluar la actividad antimicrobiana de diversas mieles frente a cepas de bacterias gram negativas y positivas, así como determinar los parámetros físico-químicos de estas mieles. Se evaluaron siete mieles de varias especies de abejas sin aguijón. Los parámetros físico-químicos evaluados fueron pH, humedad, actividad del agua, acidez, ceniza, conductividad eléctrica y color. La actividad antimicrobiana se determinó mediante pruebas de agar de difusión en disco y concentraciones inhibitorias mínimas. Encontramos que existe una relación entre los parámetros físico-químicos y la actividad antimicrobiana. La concentración mínima inhibitoria del 25% de miel fue capaz de inhibir el crecimiento de bacterias tanto grampositivas como negativas; la mayor eficacia se verificó para las especies de abejas Melipona mondury, M. quadrifasciata, Scaptotrigona bipunctata y Tetragona clavipes. En cuanto al sinergismo, Escherichia coli mantuvo su perfil de sensibilidad en relación a todas las mieles estudiadas combinadas con antimicrobianos. Un factor importante a considerar es la concentración de miel capaz de sensibilizar al microorganismo, ya que se ha demostrado que depende de la especie de abeja sin aguijón. Sin embargo, todas las mieles mostraron actividad antimicrobiana en varios métodos de análisis. Estos datos sugieren que la miel es una alternativa prometedora para sensibilizar a los microorganismos resistentes, tanto para la salud de los seres humanos como de los animales. Este estudo teve como objetivo avaliar a atividade antimicrobiana de mel de abelhas frente a cepas de bactérias gram-negativas e positivas, bem como determinar os parâmetros físico-químicos desse mel. Foram avaliadas sete amostras mel de espécies de abelhas sem ferrão. Os parâmetros físico-químicos avaliados foram pH, umidade, atividade de água, acidez, cinzas, condutividade elétrica e cor. A atividade antimicrobiana foi determinada por meio de testes de ágar de difusão em disco e concentrações inibitórias mínimas. Verificamos que existe uma relação entre os parâmetros físico-químicos e a atividade antimicrobiana. A concentração inibitória mínima de 25% de mel foi capaz de inibir o crescimento de bactérias gram-positivas e-negativas; a maior eficácia foi verificada para as espécies de abelhas Melipona mondury, M. quadrifasciata, Scaptotrigona bipunctata e Tetragona clavipes. Em relação ao sinergismo, Escherichia coli manteve seu perfil de sensibilidade em relação a todos os méis estudados associados a antimicrobianos. Um fator importante a se considerar é a concentração de mel capaz de sensibilizar o microrganismo, pois tem se mostrado dependente da espécie de abelha sem ferrão. No entanto, todas as amostras de mel apresentaram atividade antimicrobiana em diferentes métodos de análise. Esses dados sugerem que o mel é uma alternativa promissora para sensibilizar microrganismos resistentes, para a saúde de humanos e animais.

Soybeans in natura can be used in the feed of confined animals as a source of protein and energy. We measured consumption, performance, in vivo biometry, and metabolic profiles lambs fed whole grain soybean in natura. We used Ile de France breed lambs, weaned and confined in a completely randomized design with six repetitions and three treatments: Control, soybean grain (SG) 6.25%, and SG 14% (based on dry matter) in feed. The control and SG 6.25% groups showed higher average daily gain (0.296 kg/day) and total weight gain (17.57 kg) than did the SG 14% group. There were no differences among groups in terms of consumption of dry matter, protein, neutral detergent fiber, acidic detergent fiber, or feed conversion. The inclusion of soybean grain was associated with linear increases in consumption of ethereal extract, and in percentage of live weight. There were no differences among groups in terms of concentrations of studied metabolites. Regarding in vivo biometrics, the control treatment and SG 6.25% groups were superior in terms of conformation and body conditions. Inclusion of soybean whole grain in natura in the feed of confined lambs may be reduced consumption of dry matter and average daily weight gain. La soja in natura se puede utilizar en la alimentación de animales confinados como fuente de proteínas y energía. Fueron mensurados el consumo, rendimiento productivo, biometría in vivo y perfiles metabólicos de corderos alimentados con soja integral in natura. Se utilizaron corderos de raza Ile de France, destetados y confinados en un diseño completamente al azar con seis repeticiones y tres tratamientos: Control, grano de soja (SG) 6.25% y SG 14% (basado en materia seca) en el pienso. Los grupos de control y SG 6.25% mostraron una ganancia diaria promedio (0,296 kg / día) y un aumento de peso total (17,57 kg) más altos que el grupo SG 14%. No hubo diferencias entre los grupos en términos de consumo de materia seca, proteína, fibra en detergente neutro, fibra en detergente ácido y conversión alimentaria. La inclusión de grano de soja se asoció con incrementos lineales en el consumo de extracto etéreo y en porcentaje del peso vivo. No hubo diferencias entre los grupos en términos de concentraciones de metabolitos estudiados. Con respecto a la biometría in vivo, el tratamiento de control y los grupos de SG 6.25% fueron superiores en términos de conformación y condiciones corporales. La inclusión de granos integrales de soja in natura en la alimentación de corderos confinados puede reducir el consumo de materia seca y la ganancia de peso diaria promedio. A utilização do grão de soja in natura na dieta de ovinos pode ser uma alternativa na nutrição de animais confinados, como fonte de proteína e energia. Avaliar os níveis da inclusão de grão de soja inteiro in natura em dietas de cordeiros sobre o consumo, desempenho, biometria in vivo, perfil metabólico. Foram utilizados 18 cordeiros da raça Ile de France desmamados e confinados em delineamento inteiramente casualizado com seis repetições e três tratamentos: Controle, GS 6,25% e GS 14% de inclusão de grão de soja na dieta (com base na MS). O grupo Controle e GS 6,25% apresentaram maior ganho médio diário (0,296 kg/dia) e ganho de peso total (17,570 kg) em comparação ao GS 14%. Não foram observadas diferenças no consumo de matéria seca, proteína, FDN, FDA expressos em kg/dia e na conversão alimentar entre os tratamentos. A inclusão do grão de soja levou a aumento linear no consumo de extrato etéreo (EE), em kg/dia, em porcentagem do peso vivo. Não houve diferença entre os grupos quanto a concentração dos metabólitos estudados. Quanto a biometria in vivo o tratamento controle e GS 6,25% foram superiores para a conformação e condição corporal. A inclusão do grão de soja inteiro in natura em dietas para cordeiros em confinamento deve ser feita com cautela, pois pode provocar redução no consumo de matéria seca e no ganho médio diário.

This study compared the effects of unchopped and chopped hay on milk production and composition, feed digestibility, and physiology of Lacaune sheep. Eighteen ewes were stratified by parity (two or three), days of lactation (60 ± 3.7 days), and milk production (1.04 L/ewe/day), and were randomly assigned to 1 of 2 treatments: 1) Unchopped or 2) Chopped Tifton 85 hay. Corn silage and concentrate were also provided. Chopped hay ewes gave lower (P≤0.01) dry matter intake and greater (P=0.02) crude protein digestibility. No effects of treatment were detected (P≥0.38) for the digestibility of dry matter, or of neutral and acid detergent fibers. Chopped hay ewes had greater (P≤0.01) milk production (d 7 and d 12), lactation persistence, and feed efficiency. Ewes eating chopped hay had greater (P=0.03) protein concentration in their milk. However, there were no effects of treatment × day and treatments (P≥0.16) for concentrations of fat, lactose, minerals, deffated dry extract, or density. Ewes eating chopped hay presented lower (P

Resumo: Este estudo foi planejado para avaliar se a suplementação de leitões com plasma sanguíneo suíno desidratado (PSSP) no dia do nascimento tem efeito positivo sobre o metabolismo proteico e ganho de peso (GP) em leitões, bem como no desempenho na fase de creche. Selecionamos 30 leitegadas de marrãs, das quais, no mínimo um casal de leitões para cada tratamento/leitegada foi selecionado. Os tratamentos foram: TA = controle sem PSSP, TB = administração oral de 10 g de PSSP e TC = 20 g de PSSP. No nascimento, os leitões foram pesados, identificados e distribuídos aleatoriamente. Os pesos corporais também foram avaliados nos dias 7, 14 e 19. Foram coletadas amostras de sangue para medir o nível de uréia, proteína total (PT), albumina (AB) e globulina (GB). No desmame, 70 leitões de cada tratamento foram distribuídos em cinco grupos (réplicas) de 14 leitões (sete machos e sete fêmeas). Os dados de pesos (animais e consumo de ração) foram coletados no pós-desmame e também a cada troca de dieta (dias 14, 28 e 42 da fase de creche). Durante as duas primeiras semanas na maternidade, não houve efeitos dos tratamentos para todas as variáveis avaliadas. Na terceira semana, os leitões de TC apresentaram níveis séricos mais altos para as variáveis PT e GB, mas o GT foi menor nos leitões comparado ao controle. Não houve efeito dos tratamentos no GT durante todo o período lactente. Não houve efeitos residuais no desempenho da fase de creche; no entanto, os leitões tratados tiveram menos necessidade de medicamentos. A suplementação com PSSP não influenciou o desempenho zootécnico nas duas primeiras semanas ou durante todo o período lactente. O grupo TC apresentou níveis mais altos de globulina do que o grupo TA. Na fase de creche, não houve efeitos no desempenho; no entanto, houve uma maior incidência de animais doentes do grupo TA. Abstract: This research was designed to evaluate whether supplementation of piglets with spray dried porcine plasma (SDPP) on the day of birth, have positive effect on protein metabolism and weight gain (WG) in suckling piglets, as well as on performance at the nursery phase. We selected 30 litters from gilts of at least one pair of piglets for each treatment, TA= control without SDPP, TB= oral administration of 10 g of SDPP, and TC= 20 g of SDPP. At birth, the piglets were weighed, identified and randomly distributed. Body weights were also measured on days 7, 14 and 19. Blood samples were collected to measure the level of urea, total protein (TP), albumin (AB) and globulin (GB). At weaning, 70 piglets from each treatment were distributed in five groups (replicates) of 14 piglets (seven males and seven females). Weights (animals and feed intake) were taken at post-weaning and at each feed change (days 14, 28 and 42 of raising). During the first two weeks of lactation, there were no effects of the treatments for the all evaluated variables. In the third week, TC piglets had higher serum levels of TP and GB, but lower WG than did control piglets. There was no effect of treatments on WG during the entire suckling period. There were no residual effects on nursery phase performance; however, treated piglets had less need for medications. SDPP supplementation did not influence zootechnical performance in the first two weeks or during the entire suckling period. The TC group had higher globulin levels than did the TA group. In the weaning phase, there were no effects on performance; however, there was a higher incidence of diseased animals in the TA group. Resumen: Este estudio fue diseñado para evaluar si la suplementación de lechones con plasma porcino secado por atomización (PPSA) el día del nacimiento, tiene un efecto positivo sobre el metabolismo de las proteínas y el aumento de peso (AP) en lechones lactantes, así como el rendimiento en la fase de cría. Seleccionamos 30 camadas de marranas al menos un par de lechones para cada tratamiento: TA = control sin PPSA, TB = administración oral de 10 g de PPSA y TC = 20 g de PPSA. Al nacer, los lechones fueron pesados, identificados y distribuidos al azar. Los pesos corporales también se midieron los días 7, 14 y 19. Se recogieron muestras de sangre para medir el nivel de urea, proteína total (PT), albúmina (AB) y globulina (GB). Al destete, se distribuyeron 70 lechones de cada tratamiento en cinco grupos (réplicas) de 14 lechones (siete machos y siete hembras). Los pesos (animales y consumo de alimento) se tomaron después del destete y en cada cambio de alimento (días 14, 28 y 42 de crianza). Durante las primeras dos semanas de lactancia, no hubo efectos de los tratamientos para todas las variables evaluadas. En la tercera semana, los lechones TC tenían niveles séricos más altos de TP y GB, pero un AP más bajo que los lechones de control. No hubo efecto de los tratamientos sobre el AP durante todo el período de lactancia. No hubo efectos residuales en el rendimiento de la fase de vivero; sin embargo, los lechones tratados tenían menos necesidad de medicamentos. La suplementación con SDPP no influyó en el rendimiento zootécnico en las primeras dos semanas o durante todo el período de lactancia. El grupo TC tenía niveles de globulina más altos que el grupo TA. En la fase de destete, no hubo efectos en el rendimiento; sin embargo, hubo una mayor incidencia de animales enfermos en el grupo TA. Made available in DSpace on 2021-08-12T12:03:08Z (GMT). No. of bitstreams: 1 final9493.pdf: 385083 bytes, checksum: 5f62c3a2e375ab475c2eb93e9997c4f9 (MD5) Previous issue date: 2020

The objective of this study was to determine whether inoculating honey from Apis mellifera into broiler chick eggs (in ovo nutrition) during incubation would improve hatchability and performance. Initially, four hundred eggs were incubated; at 12 days of incubation, ovoscopy was performed to select the fertile eggs and divide them into three groups: control, saline and honey groups. On the 17th day of incubation we inoculated 0.2 mL of each solution directly into the amniotic cavity. After hatching, the chicks were housed in an experimental house until 28 days of age, when two birds per experimental unit were sacrificed to evaluate carcass yield. The hatchability was not affected by inoculation of saline or honey (p>0.05). However, mortality was higher in the honey and saline groups than in the control group (p0.05). These results show that the in ovo nutrition with bee honey resulted in lower production costs, that is, the birds consumed less food (feed) and had the same weight gain as other treatments. O objetivo deste estudo foi determinar se a inoculação de mel de Apis mellifera em ovos de frangos de corte (nutrição in ovo) durante a incubação melhoraria a eclodibilidade e o desempenho. Inicialmente, quatrocentos ovos foram incubados; aos 12 dias de incubação, foi realizada ovoscopia para selecionar os ovos férteis e dividi-los em três grupos: controle, solução salina e mel. No 17º dia de incubação, inoculamos 0,2 mL de cada solução diretamente na cavidade amniótica. Após a eclosão, os pintos foram alojados em um galpão experimental até os 28 dias de idade, quando duas aves por unidade experimental foram submetidas a euthanasia para avaliar o rendimento de carcaça. A eclodibilidade não foi afetada pela inoculação de soro fisiológico ou mel (p>0,05). No entanto, a mortalidade foi maior nos grupos mel e soro fisiológico do que no grupo controle (p0,05). Esses resultados mostram que a nutrição in ovo com mel de abelha resultou em menores custos de produção, ou seja, as aves consumiram menos alimento (ração) e tiveram o mesmo ganho de peso que os demais tratamentos.

Fish represents an important source of proteins and essential micronutrients for human health, and Nile tilapia (Oreochromis niloticus) is one of the most widespread aquaculture species grown worldwide to meet the growing demand for fish. In this sense, the use of supplemented diets it is important and an interesting approach to improve the growth performance, as the use of vegetable choline (VC). A total of 160 fingerlings Nile tilapia (7.10 ± 0.45 g; 6.13 ± 0.59 cm) were randomly separated into four groups in quadruplicate with 10 fish per aquarium (60 L) as follows: the group T0 (basal diet without VC supplementation), T400 (basal diet supplemented with 400 mg VC/kg of feed), (basal diet supplemented with 800 mg VC/kg of feed), and T1200 (basal diet supplemented with 1200 mg VC/kg of feed), being fed with rate of 6% of total biomass for 60 consecutive days in two equal portions and offered to fish twice a day at 9:00 am and 17:00 h. Thus, the aim of this study was to evaluate whether phosphoryl transfer network, as well as the improvement of antioxidant/oxidant status, can be considered pathways involved in the improvement of growth performance in Nile tilapia (7.10 ± 0.45 g) fed 0.0, 400, 800, and 1200 mg VC/kg of feed for 60 days. VC-supplemented diets were able to increase fish growth and feed intake after 60 days post-feeding compared to the control group. Also, fish from groups supplemented with 800 and 1200 mg VC/kg of feed presented an increase on hepatic pyruvate kinase activity compared to the control group. On the other hand, fish supplemented with 1200 mg VC/kg of feed presented an increase on hepatic glucose levels, while the hepatic lactate levels decreased compared to the control group. Additionally, VC supplementation improves the zootechnical performance. Finally, fish from groups supplemented with 800 and 1200 mg VC/kg of feed presented lower hepatic lactate dehydrogenase and lipid peroxidation levels, while the antioxidant capacity against peroxyl radicals levels were higher compared to the control group. Based on these evidences, VC supplementation can be an interesting alternative to increase growth performance of Nile tilapia fingerlings especially at levels of 800 and 1200 mg VC/kg of feed, which were able to improve the hepatic energy metabolism and antioxidant/oxidant status and subsequently increased the fish growth performance.

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.

The aim of this study was to determine whether the addition of curcumin (free and encapsulated) to chick feed would minimize the negative effects on health and performance caused by daily intake of fumonisin. We used 50 birds, divided into five treatments: CP, basal diet with 600 mg/kg of fumonisin, with antibiotic and coccidiostatic agent; CU, 600 mg/kg of fumonisin and 50 mg/kg of curcumin; NC5, feed with 600 mg/kg of fumonisin and 5 mg of nano-curcumin/kg of feed; NC10, feed with 600 mg/kg of fumonisin and 10 mg of nano-curcumin/kg of feed; and CN, fumonisin-free diet, with antibiotic and coccidiostatic. We measured weights, weight gain, and serum biochemistry, as well as antioxidant and oxidant activities. Lower body weight and weight gain were observed in chicks that received feed with fumonisin; curcumin did not minimize this negative effect. Lower glucose and triglyceride levels were also observed in the NC10 group, while the highest cholesterol levels were observed in all groups of birds that consumed fumonisin compared to the CN group. Uric acid levels were significantly lower in CP than in CN. Levels of liver enzymes were higher in CP than in CN. The highest levels of thiobarbituric acid reactive substances were found in CP and CU, whereas ROS was higher in CU compared to CN. Superoxide dismutase activity was significantly lower in CP, while glutathione S-transferase activity was higher in the CP group. Catalase activity was lower in groups of birds that consumed fumonisin compared to CN. Taken together, these findings suggest that intake of curcumin-loaded nanocapsules (10 mg/kg) had hepaprotective and antioxidant effects in chicks artificially intoxicated with fumonisin, minimizing the negative effects caused by this mycotoxin.

This study evaluated the effect of different amounts of marine microalgae (Schizochytrium limacinum) via milk on performance, serum constituents and antioxidant indicators of dairy calves. Twenty-four Holstein calves were distributed to one of four treatments (six replicates) according to a completely randomized design: a control treatment (without microalgae), or microalgae supplementation at 6, 12 and 18 grams of microalgae per day supplied via milk. Two blood samples were taken, at 21 and 42 days of the experiment, to measure activities of antioxidant enzymes: glutathione S-transferase (GST), catalase (CAT), superoxide dismutase (SOD) as well as levels of reactive oxygen species (ROS). The supplementation of increased amount of microalgae did not influence dry matter intake (DMI) and performance (P>0.05). At the 21 days of life, there was a linear increase in serum protein (P = 0.03), globulin (P = 0.006) and in SOD activity (P = 0.05). Linear reduction in ROS (P = 0.001) and a quadratic effect for heavy immunoglobulin G (IgG) (P = 0.02) were observed as the microalgae amounts increased. Close to weaning, at 42 days, serum globulin (P = 0.003), GST (P = 0.002) and SOD (P = 0.02) linearly increased with the inclusion of microalgae. Conversely, enzymatic activity of CAT showed a quadratic effect (P = 0.05). Supplementation of microalgae via milk did not influence intake and performance dairy calves. However, the use of microalgae as a feed additive to dairy calves at amount of 6 g/d is recommended due to its stimulation of antioxidant enzymes and reduction on serum levels of free radicals. Este estudio evaluó el efecto de diferentes cantidades de microalgas marinas (Schizochytrium limacinum) a través de la leche sobre el rendimiento, los componentes del suero y los indicadores antioxidantes de los terneros lecheros. Veinticuatro terneros Holstein se distribuyeron a uno de cuatro tratamientos (seis repeticiones) de acuerdo con un diseño completamente al azar: un tratamiento de control (sin microalgas) o suplementos de microalgas a 6, 12 y 18 gramos de microalgas por día suministrados a través de la leche. Se tomaron dos muestras de sangre, a los 21 y 42 días del experimento, para medir las actividades de las enzimas antioxidantes: glutatión S-transferasa (GST), catalasa (CAT), superóxido dismutasa (SOD), así como los niveles de especies reactivas de oxígeno (ROS). La suplementación de una mayor cantidad de microalgas no influyó en la ingesta de materia seca (DMI) y el rendimiento (P> 0.05). A los 21 días de vida, hubo un aumento lineal en la proteína sérica (P = 0.03), la globulina (P = 0.006) y en la actividad de SOD (P = 0.05). Se observó una reducción lineal en ROS (P = 0.001) y un efecto cuadrático para la inmunoglobulina G pesada (IgG) (P = 0.02) a medida que aumentaron las cantidades de microalgas. Cerca del destete, a los 42 días, la globulina sérica (P = 0.003), GST (P = 0.002) y SOD (P = 0.02) aumentaron linealmente con la inclusión de microalgas. Por el contrario, la actividad enzimática de CAT mostró un efecto cuadrático (P = 0.05). La suplementación de microalgas a través de la leche no influyó en la ingesta y el rendimiento de los terneros lecheros. Sin embargo, se recomienda el uso de microalgas como aditivo alimenticio para terneros lecheros en una cantidad de 6 g por día debido a su estimulación de enzimas antioxidantes y la reducción de los niveles séricos de radicales libres. Objetivou-se avaliar se a adição de diferentes níveis de microalgas marinhas (Schizochytrium limacinum) via leite tem efeitos positivos sobre saúde e desempenho de bezerras leiteiras. Foram utilizadas 24 bezerras da raça holandesa, distribuídas em delineamento inteiramente casualizado: sem adição de microalgas, 6 gramas de microalga por dia, 12 gramas de microalga por dia e 18 gramas de microalga por dia fornecidas via leite. Foram realizadas duas coletas de sangue, aos 24 e 42 dias de vida, para avaliar enzimas antioxidantes: glutationa S-transferase (GST), catalase (CAT), superóxido dismutase (SOD) e espécies reativas ao oxigênio (ERO). A suplementação de quantidades crescentes de microalgas não alterou a ingestão de matéria seca (IMS) e desempenho (P>0,05). Aos 21 dias de vida, houve aumento linear para proteína (P = 0,03), globulina (P = 0,006) e SOD (P = 0,05). Observou-se redução linear para ERO (P = 0,001) e quadrática para imunoglobulina de cadeia pesada (IgG) (P = 0,02) quando as doses de microalgas aumentaram. Próximo ao desmame, aos 42 dias, a inclusão de microalgas aumentou de forma linear os níveis de globulina sérica (P = 0,003), GST (P = 0,002) e SOD (P = 0,02). Por outro lado, atividade enzimática da CAT apresentou efeito quadrático (P = 0,05). A suplementação com microalgas via leite não altera a ingestão e desempenho de bezerras leiteiras. No entanto, o uso como aditivo alimentar na dose de 6 g ao dia é recomendada devido à estimulação das enzimas antioxidantes e redução do níveis séricos de radicais livres.

Purinergic signaling is linked to neurodegenerative and proinflammatory damage during pathological conditions such as hypoxia, but involvement of this pathway in brain damage in fish exposed to environmental hypoxia remains unknown, and we propose dietary supplementation with caffeine in order to improve the immune response. Therefore, the aim of the study was to evaluate whether the enzymatic purinergic signaling pathway is associated with inflammatory brain damage in Nile tilapia (Oreochromis niloticus) exposed to environmental hypoxia and whether dietary supplementation with caffeine (5% and 8%) can prevent these changes in purinergic signaling. Animals were randomly divided into six groups (A–F, n = 6 per group, in triplicate), as follows: groups A–C were submitted to normoxia, while groups D–F were submitted to hypoxia. Groups A and D received the basal diet, while groups B and D and groups C and F received a diet containing 5% and 8% caffeine, respectively, and fed with their respective diets for 21 days. After 21 days, aeration was disconnected (groups D–F) and the dissolved oxygen levels were maintained as follows: group A (6.55 ± 0.23 mg/L), group B (6.51 ± 0.24 mg/L), group C (6.58 ± 0.22 mg/L), group D (1.23 ± 0.11 mg/L), group E (1.20 ± 0.15 mg/L), and group F (1.18 ± 0.13 mg/L). Cerebral triphosphate diphosphohydrolase (NTPDase) using adenosine triphosphate (ATP) as a substrate and 5′-nucleotidase activities decreased in fish exposed to 72 h of hypoxia compared with the normoxia group, while adenosine deaminase (ADA) activity and levels of nitric oxide (NOx) metabolites were higher. Dietary supplementation with 5% and 8% caffeine prevented all alterations elicited by hypoxia, with the exception of ADA activity in the case of 5% caffeine. Based on this evidence, our findings reveal that nucleotide/nucleoside hydrolysis is modified in the brains of fish exposed to 72 h of hypoxia, contributing to inflammatory damage, which apparently is mediated by excessive ATP content in the extracellular medium and by excessive NOx production. Also, the use of a diet containing 5% and 8% caffeine prevented these alterations (except 5% of dietary caffeine on ADA activity) and can be considered an interesting approach to preventing the impairment of immune and inflammatory responses elicited by hypoxia, principally the inclusion of 8% caffeine.

Adenosine triphosphate (ATP) and adenosine (Ado) are the main molecules of the purinergic signalling involved in toxicological mechanisms induced by pesticides. These molecules participate in the regulation of immune and inflammatory responses due to their interaction with purinereceptors in the extracellular medium. In the literature, the ATP and Ado are related with pro-inflammatory and immunomodulatory effects, respectively. Thus, the aim of this study was to evaluate whether the purinergic signalling can be considered a potential target for trichlorfon-induced inflammation and impairment of the immune response. Seric and splenic triphosphate diphosphohydrolase (NTPDase) activities using ATP as substrate were reduced at 22 mg/L after 48 h of exposure compared to the control group, while NTPDase activity using ADP as substrate was lower only in the serum of silver catfish at the same period. On the other hand, seric and splenic adenosine deaminase (ADA) activities and metabolites of nitric oxide (NOx) levels were increased at 22 mg/L after 48 h of exposure compared to the control group. The enzymatic activity of the purinergic signalling did not return to control levels after 48 h of recovery period in trichlorfon-free water. Based on these evidences, we concluded that trichlorfon interferes with the purinergic signalling, causing impairment of the immune and inflammatory responses by reducing ATP hydrolyses, possible leading to its accumulation in the extracellular environment. In addition, there is an increase in Ado desamination and possible reduction of it in the extracellular medium, leading to a self-sustained pro-inflammatory deleterious cycle. In summary, the purinergic signalling can be considered a potential target for trichlorfon-induced inflammation and impairment of the immune response in freshwater silver catfish at exposure to 22 mg/L.

The aim of this study was to evaluate whether a phytogenic feed additive (PFA) based on essential oils such as carvacrol, thymol and cinnamic aldehyde, could be considered a replacement for antimicrobials used as growth promoters in broiler chickens, as well as to investigate its effect on total bacterial count, biochemical profiles, meat quality and meat fatty acid profile. A total of 240 broiler chicks were randomly distributed into 4 groups with 4 replicates of 15 animals each, as follow: T1 (basal diet only; the control group), T2 (basal diet supplemented with zinc bacitracin), T3 (basal diet with 0.5% of the PFA), T4 (basal diet with 1.0% of the PFA). The addition of 0.5% of the PFA improved live body weight of supplemented birds compared to the control group at 35 and 42 days of age, while the total bacterial count in the environment was reduced when 1.0% of the PFA was used. In addition, intestinal villi height and crypt depth suffered variations during the entire experiment in birds treated with both concentrations of the PFA and zinc bacitracin. Total erythrocyte counts were higher on days 14, 28 and 42 in both treated groups (PFA) compared to the control group, as well as hemoglobin content on days 28 and 42. On the other hand, leukocyte counts were lower on days 14, 28 and 42 due to reduced lymphocyte counts in both PFA treated groups compared to the control group. Serum levels of alanine aminotransferase (ALT) were lower in broilers fed with either concentration of PFA on day 14 of life, and the same was observed regarding aspartate aminotransferase (AST) in broiler treated with 0.5% of the PFA. Also, total protein and globulin levels were lower on days 14 and 28 in groups fed with phytogenic compared to the control group. Regarding meat quality, breast meat showed higher red intensity and shear force in groups fed with both concentrations of phytogenic compared to the control group, while weight loss by cooking was lower. Finally, 1.0% of phytogenic showed lower docosadienoic acid (C22:2) content in breast meat. In conclusion, results showed that the use of PFAs based on carvacrol and thymol may be considered an interesting alternative to increase broilers performance, replacing the use of antimicrobials as growth promoters, as well as an interesting alternative to reduce the total bacterial count in the environment of broiler chickens. Moreover, the diet containing phytogenic also showed hepaprotective effects but deserves attention regarding possible alterations on the immune response.

The objective of this study was to evaluate if there were alterations in hematological variables of fish after transportation and exhibition at an agricultural fair. We first collected blood samples from 12 fish of the species Prochilodus lineatus (curimbata) at the Goio-En research institute; then, these animals were transported (60 km) to the agricultural fair, where they stayed for 3 days in glass aquariums exposed to the visitors of the event. On the return of the fish to the institute, we collected blood again. We performed hematological analyses, measuring hematocrit, hemoglobin concentration, and total erythrocyte count in the harvested material. We also determined mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration. Total leukocyte counts and leukocyte differential performed on blood smears. There were no differences in hematocrit, hemoglobin, mean corpuscular hemoglobin, leukocyte, lymphocyte, neutrophil, and monocyte concentration (P > 0.05) before and after the fair. However, the number of erythrocytes and thrombocytes were lower after the fish show (P

Several evidences have related the biochemical and pharmacological properties of thymol, but the possible neurotoxic effects of this compound remain unknown and not evaluated. Thus, the purpose of this study was to evaluate whether intake of thymol in different doses (10, 20 and 40 mg/kg) induce neurotoxicity and behavioral alterations using mice as experimental model, as well as the involvement of blood-brain barrier (BBB) and brain neurotransmitters in these alterations. Thymol (20 and 40 mg/kg) significantly decrease latency time to inhibitory avoidance task when compared to control group, indicating a memory loss after 30 days of oral treatment. Also, thymol (20 and 40 mg/kg) induced a significant increase on BBB permeability to Evan’s blue dye when compared to control group, which is an indicative of BBB breakdown. Moreover, a significant increase of brain acetylcholinesterase (AChE) was observed in mice treated with 40 mg/kg of thymol, while the activity of sodium-potassium pump (Na+, K+-ATPase) was inhibited in mice treated with 20 and 40 mg/kg thymol when compared to control group. Finally, mice that received 20 and 40 mg/kg thymol showed a significant increase on cerebral reactive oxygen species (ROS) levels and cerebral xanthine oxidase (XO) activity compared to control group. Based on these evidences, the rupture of BBB can be considered an important pathway linked in thymol-induced memory loss. Also, the augmentation of brain ROS levels elicited by increase on XO activity may be a via involved in the damage to BBB, and an oxidative pathway that impairs the activity of brain neurotransmitters, as AChE and Na+, K+-ATPase. In summary, the dose of 10 mg/kg thymol can be safe and without neurotoxic effects in a period of 30 days of intake.

Thymol is a phytochemical component present in many plants used as food additive in order to promote animal growth due to its several biological properties. However, possible side effects of thymol remain poorly known limited to few reports. In this sense, we evaluated the enzymes of the purinergic signaling such as, ectonucleoside triphosphate diphosphohydrolase (NTPDase), 5'-nucleotidase and adenosine deaminase (ADA), that play an important role on toxicity induced by excessive adenosine triphosphate (ATP) content in the extracellular environment. Thus, the aim of this study was to evaluate whether purinergic signaling could be considered a potential target of thymol-induced inflammation, and the toxicity in tissues and immune cells of mice after thymol administration. NTPDase activity (ATP as substrate) in serum, spleen, and splenic lymphocytes was lower after 30 days of oral treatment at doses of 10, 20, and 40 mg/kg of thymol, while ADA activity was stimulated at 20 and 40 mg/kg. No differences were observed between groups regarding NTPDase (ADP as substrate) and 5'-nucleotidase activities in all evaluated tissues. Based on these evidences, adenine nucleotide hydrolysis is modified in serum, spleen, and splenic lymphocytes of mice treated with thymol, contributing to inflammation and toxicity by a reduction on ATP hydrolyses and its possible accumulation in the extracellular medium and increased Ado desamination and its possible reduction in the extracellular environment, leading to a self-sustained pro-inflammatory deleterious cycle. In summary, all tested thymol concentrations induced inflammation and toxicity in tissues and immune cells of treated mice.

Rangelia vitalii is the etiologic agent of canine rangeliosis, a severe piroplasmosis that affects domestic dogs in Brazil, Uruguay and Argentina. While R. vitalii is one of the most pathogenic tick-borne pathogens for dogs in the world, its tick vector has remained unknown. The present study evaluated the vector competence of Rhipicephalus sanguineus sensu lato (both tropical and temperate species), Amblyomma aureolatum, Amblyomma ovale, Amblyomma tigrinum, and Amblyomma sculptum for R. vitalii. These six tick species were selected for the study because they comprise the main tick species infesting dogs within the distribution area of canine rangeliosis in South America. Acquisition feeding of the above six tick species was performed on domestic dogs showing clinical signs of canine rangeliosis, after being experimentally infected through intravenous inoculation or infestation with R. vitalii-infected ticks. Thereafter, engorged ticks were evaluated for transstadial and transovarial passages of R. vitalii through molecular analysis after molting or oviposition and egg hatching. The resultant ticks were evaluated for their competence to transmit R. vitalii to susceptible dogs. Among the six tick species, only A. aureolatum was able to acquire and perpetuate R. vitalii by transstadial and transovarial passages, as demonstrated by >5% infection rates of ticks after hatching or molting. When exposed to transmission feeding, only A. aureolatum ticks were competent to transmit R. vitalii to dogs, which became severely ill, and the results confirmed by molecular methods and blood smear examination to have acquired rangeliosis. Results of the present study, coupled with epidemiological data, indicate that A. aureolatum is a natural vector of R. vitalii. Our results also indicate that R. vitalii is the first Piroplasmorida agent to be transovarially transmitted in Amblyomma ticks.