Addition of Curcumin in Dairy Sheep Diet in the Control of Subclinical Mastitis

Background: Curcumin, a polyphenol derived from rhizome of Curcuma longa, has been successfully used in animals and humans due to its antibacterial and anti-inflammatory properties. Recently, a study demonstrated that curcumin ointment reduced pain, mammary tension and erythema in humans with mastitis 72 h after the treatment. Mastitis in sheep is an important problem due to restrictions associated to the use of antibiotics, affecting milk production and becoming a public health issue. Thus, the aim of this study was to verify whether the addition of curcumin in the diet of six female sheep with subclinical mastitis could have curative efficacy and benefits to animal health.Cases: Six Lacaune sheep with subclinical mastitis that persisted for more than thirty days after the parturition were used. The animals received feed containing the curcumin (60 mg/animal/day) during 10 days. On days 0 and 10 of the experiment, total blood was collected to perform the hemogram and serum biochemistry (triglycerides, cholesterol, glucose, total protein (TP), albumin, globulin, aspartate aminotransferase (AST) and urea), as well as individual milk samples for centesimal composition, somatic cell count (SCC) and total bacterial count (TBC). Total leukocytes and erythrocytes, as well as hemoglobin content, were obtained using the hematological counter CELM-CC-550. Blood smears were stained by theRomanowsky method and used for leukocytes differentiation. After ten days of treatment, five sheep were negative in the California Mastitis Test (CMT). Total erythrocytes increased, while total leukocytes decreased after the treatment. Seric levels of AST, triglycerides, cholesterol and globulins decreased, while seric levels of albumin and urea increased after tendays of treatment. No differences were observed regarding milk production and composition, as well as for SCC and TBC on days 0 and 10 post treatment.Discussion: The use curcumin to prevent or to delay the occurrence of subclinical mastitis in ewes was first tested. In an experiment of lipopolysaccharide-induced mastitis in rats, researchers identified receptors type Toll 4 which are related with the induction of the synthesis of pro-inflammatory cytokines, as tumor necrosis factor alpha, interleukin-6 and 1β, molecules involved in the migration of leukocytes to the site of inflammation. According to literature, curcumin was able to reduce theactivity of myeloperoxidase in the mammary gland, an enzyme used as indicator of neutrophil infiltration, demonstrating the potent anti-inflammatory effect of curcumin. This effect can be associated to the release of interleukin-8, a cytokine that promotes the migration of neutrophils, lymphocytes and eosinophils into the site of infection, i.e., the mammary gland. It is important to highlight that neutrophils, due to its capacity to release inflammatory mediators and to delay apoptosis, has the potential to maintain a permanent inflammatory process, effects that were prevented or reduced by curcumin, specially the apoptotic and cytotoxic effects. It is important to emphasize the direct effect of curcumin on bacteria present in the mammary gland, since it exerts potent bactericidal effect. In summary, the use of curcumin in sheep with subclinical mastitis exerted beneficial effects in the dose of 60 mg/sheep/day, since it showed 83.3% of curative efficacy in the test CMT. This effect of curcumin can be direct or indirect, since the curcumin provided in the diet showed anti-inflammatory properties byreducing oxidation reactions and cell damage. Overall, the addition of curcumin in sheep diet improved the hematological and biochemical variables after only ten days of treatment. Future studies should be performed to clarify and determine whether the curcumin provided in diet can be an alternative to treat clinical or subclinical mastitis.Keywords: curcumin, mammary infection, immunity, biochemistry parameters, ovine

Experimental Infection by Brucella ovis: Changes in NTPDase, 5'-Nucleotidase and Acetylcholinesterase Associated Cerebral Oxidative Stres

Background: Changes in purinergic and cholinergic signaling have been demonstrated in various pathologies associated with inflammation; however, the changes in brucellosis caused by the Gram-negative coccobacillus Brucella ovis are not known. B. ovis is generally asymptomatic in sheep. Hepatosplenomegaly has been described in B. ovis, a non-zoonotic species, characterized by an extravascular inflammatory response. Purinergic system enzymes are closely involved with the modulation of the immune system, pro- and anti-inflammatory events. The objective of this study was to investigate the role of ectonucleotidases and cholinesterase’s in the brains of mice experimentally infected with B. ovis.Materials, Methods & Results: Forty-eight animals were divided into two groups: control (n = 24) and infected (n = 24). In group infected, 100 µL containing 1.3 x 107 UFC B. ovis /mL via intraperitoneal was used in inoculation. The brains were collected from the animals on days 7, 15, 30 and 60 post-infection (PI). We measured levels of TBARS (substances reactive to thiobarbituric acid) and ROS (reactive oxygen species) in the brain. The activity of NTPDase (using ATP and ADP as substrate) and 5'-nucleotidase (using AMP as substrate) were evaluated in brain in addition to histopathological analysis. No histopathological lesions were observed in the control group nor the infected group at days 7, 15, and 30 PI. However,multifocal areas with moderate microgliosis and inflammatory infiltrates in the cerebral cortex were observed at day 60 PI in the infected animals. B. ovis DNA was detected in brain. During the course of infection, B. ovis caused greater lipid peroxidation in the brains of infected animals than in the control group at day 60PI. No significant results were observed at 7, 15 or day 30 PI. Similarly, there was significantly more reactive oxygen species at day 60 PI in brains of infected animals than in the control group. NTPDase activity (using ATP and AMP as substrate) was lower at days 7 and 15 PI in infected animals than in control. However, during the course of infection there was an increase in NTPDase activity at day 60 PI in the infected group. The infected animals showed a decrease of 5´-nucleotidase (AMP as substrate) activity at days 7 and 30 PI. On the other hand, 5´-nucleotidase activity was greater on day 60 PI in the experimental group than in the control. The results suggest that nucleotide hydrolysis was low in the acute phase (up to day 30 PI) due to the decrease of NTPDase and 5´-nucleotidase activities. After day 60 PI, there was a reversal in enzyme activities, probably with concomitant increase of extracellular nucleotides. AChE activity in brain on days 30 and 60 PI compared to control.Discussion: Among the functions of NTPDase are inhibition of platelet aggregation, vascular homeostasis, modulation of inflammation and immune response, all via its regulation of extracellular concentrations of ATP, a pro-inflammatory molecule. E-NTPDase plays an important role in controlling lymphocyte function, including antigen recognition and activation of cytotoxic T cell effector functions, as well as the generation of signals. The enzyme E-5´-nucleotidase also exerts non-enzymatic functions, including induction of intracellular signaling and mediation of cell-cell adhesion and cell-matrix and migration. Levels of acetylcholine are regulated by cholinesterase enzymes that are present in cholinergic and noncholinergic tissues, as the acetylcholinesterase (AChE) is a membrane-bound enzyme, primarily found in the brain and cholinergic neurons, where it participates in the structural regulation of postsynaptic differentiation. The results demonstrated that the chronicity of infection by B. ovis causes oxidative damage and inflammation in the brain, as well as modulation of ectonucleotidases and AChE activities

The Protective Effects of an Adsorbent against Oxidative Stress in Quails Fed Aflatoxin-Contaminated Diet

Background: Contamination of crops with aflatoxin is considered a serious global threat to food safety, since potent carcinogenic, teratogenic, mutagenic and immunosuppressive effects of aflatoxins are well recognized. Recently, the use of adsorbents has been linked with protective effects against oxidative stress in several diseases. Thus, the aim of this study was to assess the occurrence of oxidative stress in quails (Coturnix coturnix) fed with aflatoxin-contaminated diet, as well as the protective effect of an adsorbent.Materials, Methods & Results: Twenty-eight quails were divided into four groups (n = 7): diet without additives (control; the group A), diet and adsorbent containing aluminosilicates (the group B), aflatoxin-contaminated diet (200 ppb) (the group C), and aflatoxin-contaminated diet (200 ppb) and adsorbent containing aluminosilicates (the group D). The composition of the adsorbent containing aluminosilicates was 0.3% based on yeast cell wall, silymarin, and bentonite. The animals received feed and water ad libitum during 20 days. At the end of the experimental period, total blood was collected by cardiac puncture in tubes without anticoagulant to obtain serum (centrifuged at 3500 g during 10 min) for later determination of biochemical parameters. The liver was placed in a solution of Tris–HCl 10 mM, pH 7.4 for TBARS (Thiobarbituric acid reactive substances), ROS (Reactive oxygen species), SOD (Superoxide dismutase) and CAT (Catalase) analysis. The hepatic tissue was gently homogenized in a glass potter in specific buffer, homogenated, and centrifuged at 10.000 g at 4ºC for 10 min to yield a supernatant (S1) used for analyses. Homogenate aliquots were stored at -80°C until utilization. Fragments of liver and intestine (5 cm) were collected for histopathological analyses. Between days 15 to 20 of the experiment, group C quails showed clinical signs, such as apathy, creepy feathers and reduced feed intake. At day 20 of experiment, macroscopically, the liver of quails belonging to the group C showed greenish yellow color differently from the other groups. Microscopically, no alterations were observed in the liver of animals in groups A and B. Severe diffuse microvacuolar degeneration (hydropic) of hepatocytes and small foci of necrosis in the liver were observed in the group C, as observed in the group D, but in a more moderate degree to microvacuolar degeneration. Seric total protein, albumin, globulin and uric acid levels decreased in the group C and D. The levels of alanine aminotransferase (ALT) increased in the group C, and the treatment with adsorbent was able to avoid this increment. Seric and hepatic reactive oxygen species and TBARS increased in the group C, and the treatment with adsorbent reduced theses parameters in the group D. Catalase (CAT) activity decreased, while ALA-D increased in the group C. The treatment with adsorbent was able to prevent CAT activity decrease, but it did not prevent the increase in ALA-D activity.Discussion: Aflatoxins are considered one of the most important problems in poultry production causing high economic losses to producers. In this study, the use of adsorbent showed a protective effect to hepatic tissue, minimizing histopathological lesions, as well as by preventing lipid peroxidation and exacerbated production of free radicals. Based on this data, aflatoxin intoxication causes hepatic oxidative stress that contributes directly to disease pathogenesis, and the addition of an adsorbent containing 0.3% based on bentonite, yeast cell wall and silymarin may be considered a new approach to prevent cellular and hepatic damage caused by aflatoxins

Spray-dried porcine plasma added to diets contaminated with aflatoxins and fumonisins shows beneficial effects to piglet health

This study was aimed to analyze the effects of spray-dried porcine plasma (SDPP) on the health of post weaning piglets challenged with diets contaminated with aflatoxins and fumonisins. Fifty-six male piglets (7.15 ± 0.61 kg) were allocated in four groups: CTL group received a regular diet; SDPP group received a regular diet and 6% SDPP; MYC group received a diet containing 300 μg/kg aflatoxins and 8,000 μg/kg fumonisins; group MYC+SDPP received 300 μg/kg aflatoxins, 8,000 μg/kg fumonisins and 6% SDPP. The animals that received the experimental diet containing mycotoxins (MYC group) had lower weight gain at the end of the experiment compared to the other treatments. Animals receiving SDPP showed decreased urea levels throughout the experiment (P<0.05). Animals from MYC group presented an increased on reactive oxygen species (ROS) and thiobarbituric acid reactive substances (TBARS) levels and decreased catalase activity (P<0.05). In contrast, SDPP prevented the increase of ROS and TBARS and stimulated superoxide dismutase activity (P<0.05). In conclusion, diet contaminated with mycotoxins (group MYC) caused subclinical intoxication in the piglets, as observed by the increase on free radical’s production and lipid peroxidation. Conversely, SDPP presented a protective effect, minimizing the effects of oxidative stress caused by aflatoxins and fumonisins ingestion

Safety profile, antimicrobial and antibiofilm activities of a nanostructured lipid carrier containing oil and butter from Astrocaryum vulgare: in vitro studies

Ethnopharmacological relevance: Tucumã (Astrocaryum vulgare)is a fruit native to the Amazon region. Extracts from the peel and pulp are thought of as promising treatments for bacterial infections. The primary constituents of Tucumã oil and butter possess unsaturated carbon chains that are susceptible to oxidation by light or heat. The oils have high volatility and low aqueous solubility that limits their use without a vehicle. Nanotechnology refers to techniques to solve these problems. Nanostructured lipid carriers (NLC), for example, protect fixed oils degradation by heat or light, as well as from oxidation and evaporation, ensuring greater stability and function, thereby prolonging the useful life of the final product. Study objectives: The objective of this study was to evaluate the hemolytic, cytotoxic, antimicrobial and antibiofilm properties of an NLC containing Tucumã butter and oil soasto improve the solubility and photosensitivity of the compounds, generating better pharmacological efficacy. Materials and methods: The NLC was assessed for stability for 60 days. The cytotoxicity of nanoparticles in peripheral blood mononucleated cells was determined in culture using assays for cell viability, DNA damage, oxidative metabolism and damage to human erythrocytes. Antimicrobial activity was determined using the broth microdilution technique and antibiofilm activity according to standardized protocols. Results: The Tucumã NLC remained stable throughout the evaluated period, with pH between 5.22–5.35, monodisperse distribution (PDI&lt;0.3) and average particle size of 170.7 ± 3nm. Cytotoxicity studies revealed that the NLC is safe and modulates inflammatory processes, demonstrated by increased cell viability and nitric oxide levels. There was low hemolytic activity of the NLC against human erythrocytes almost concentrations tested. Conclusion: Taken together, the data suggest that NLC containing Tucumã oil and butter showed antimicrobial and antibiofilm activity against organisms that cause morbidity and mortality in humans. They may be alternative solutions to public health problems related to bacterial infections

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

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

Atividade in vitro da secreção de Lucilia cuprina frente à Leishmania amazonensis, Ttrypanosoma cruzi e linhagens celulares / In vitro activity of larval secretions from Lucilia cuprina against Leishmania amazonensis, Trypanosoma cruzi and cell lines

As pesquisas com produtos naturais representam um valioso recurso, pois são fonte de compostos biologicamente ativos que podem ter atividade contra uma variedade de patógenos que afetam a saúde humana e animal. Os dípteros da espécie Lucilia cuprina, pertencente à família Calliphoridae, conhecidos como moscas varejeiras, são cosmopolitas e têm importante função na natureza devido ao desenvolvimento de suas larvas em matéria orgânica em decomposição. Alguns estudos chegaram à conclusão de que a produção de enzimas digestivas secretadas pelas larvas são importantes para o estabelecimento e  sobrevivência desses insetos no hospedeiro e no ambiente  devido a sua  secreção ter ação bactericida, antiparasitária e antifúngica e ao hábito alimentar necrobiontófago, esses dipteros têm sido utilizados em bioterapia e pesquisas contra vários patógenos, entretanto, são poucos os estudos com foco na secreção desta espécie. Portanto, o objetivo deste trabalho foi verificar a atividade da secreção larval in vitro de L. cuprina, extraída de larvas L3 em diferentes concentrações por um período de 24 horas, frente à Leishmania amazonensis, Trypanosoma cruzi, linhagem tumoral de melanoma B16-F10 e HFF-1 de fibroblasto humano. Evidenciou-se uma redução na viabilidade de promastigotas de L. amazonensis nos tratamentos com secreção larval de 0,25%, 1% e 2% (viabilidade reduzida para 43,33%, 60% e 37,33% respectivamente). No ensaio com T. cruzi as concentrações 0,25% e 0,5% de secreção larval foi ativa mostrando diminuição de viabilidade de epimastigotas para 35,28% e 43,07%. A secreção larval mostrou capacidade citotóxica para as células tumorais. A concentração de 2% foi significativa em todos os ensaios, MTT, Vermelho Neutro (NR) e Crystal Violeta (VC), com redução de viabilidade para 79,3%, 85% e 85,6%, respectivamente. O tratamento a 1% mostrou diferença significativa tanto para NR (viabilidade de 74%) como VC (viabilidade de 83,6%). A concentração de 0,5% no NR diminuiu viabilidade para 80,6%. Nos ensaios com linhagem não tumoral a secreção mostrou baixa toxicidade, demonstrando segurança. O ensaio fluorimétrico realizado para quantificação de dsDNA (Picogreen) indicou um aumento de 14% à 25% de DNA extracelular. Os resultados fluorimétricos com o marcador 2,7’-diclorofluoresceína (DCFH-DA) mostraram um aumento de 20% e 22% de espécies reativas nas concentrações de 0,25 e 2%, sendo indicativo de um possível mecanismo de morte celular por radicais livres. Os resultados revelam que a secreção larval de L. cuprina, em diferentes concentrações, demonstrou atividade antiprotozoária e citotóxica na linhagem tumoral

Resveratrol and resveratrol-hydroxypropyl-β-cyclodextrin complex recovered the changes of creatine kinase and Na+, K+-ATPase activities found in the spleen from streptozotocin-induced diabetic rats

Type 1 diabetes (T1D) is the result of the selective destruction of the pancreatic β-cells by T cells of the immune system. Although spleen is a secondary lymphoid organ, it is also involved in the T1D pathogenesis. However, the alterations in a variety of cellular processes of this disease need to be further understood. We aimed to analyze the benefits of resveratrol, and its complexed form on diabetic complications in the spleen of rats. To this end, we investigated important enzymes of phosphoryl transfer network, and Na+, K+-ATPase activity. Wistar rats were divided into non-diabetic groups: Control, Ethanol, Resveratrol, Hydroxypropyl-β-cyclodextrin, Resveratrol-hydroxypropyl-β-cyclodextrin, and diabetic groups with the same treatments. Diabetes was induced by a single dose of 60 mg/kg of streptozocin intraperitoneally, and treatments by intragastric gavage once daily for 60 days. Hyperglycemia reduced creatine kinase activity, which was reversed by the administration of resveratrol. Na+, K+-ATPase activity was greatly affected, but it was reversed by resveratrol and resveratrol-hydroxypropyl-β-cyclodextrin. This suggest an energetic imbalance in the spleen of diabetic rats, and in case this also occurs in the diabetic patients, it is possible that resveratrol supplementation could be beneficial to the better functioning of the spleen in diabetic patients