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

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

Nursery pigs fed with feed contaminated by aflatoxin B1 (Aspergillus flavus) and anti-mycotoxin blend: Pathogenesis and negative impact on animal health and weight gain.

The present study aimed to evaluate whether a moderate dose of aflatoxin B1 in pigs' diet interferes with pigs' growth and health in the nursery phase and whether an anti-mycotoxin mixture minimizes the adverse effects of the toxin. One blend with Saccharomyces cerevisiae lysate, zeolite, silicon dioxide, propylene glycol, Carduus marianus extract, soy lecithin, and carbonate was used as an anti-mycotoxin. Four treatments, with six repetitions per treatment and three pigs/pen: Afla0-AntiMyc0 - negative control (without aflatoxin); Afla500-AntiMyc0 - positive control (500 ppb of aflatoxin); Afla0-AntiMyc1000 - 1000 mg/kg of anti-mycotoxin blend; Afla500-AntiMyc1000 - 500 ppb aflatoxin +1000 mg/kg of anti-mycotoxin blend. It was observed that pigs in the positive control (Afla500-AntiMyc0) had lower body weight and weight gain when compared to the other treatments during the experimental period. Also, pigs from Afla500-AntiMyc0 had lower feed intake between days 1-20 and 1 to 30 than Afla0-AntiMyc0. The pigs from Afla500-AntiMyc0 had higher levels of liver enzymes aspartate aminotransferase and alanine aminotransferase compared to other treatments. The pigs from Afla500-AntiMyc0 had higher villus height than the other treatments, while the folded size was smaller in this treatment. Crypts were deeper in the intestines of pigs in both treatments that consumed aflatoxin. In general, it is concluded that the intake of aflatoxin B1 by nursery pigs has negative impacts on the health and, consequently, the animals' growth performance; however, the addition of the contaminated feed with an anti-mycotoxin blend was able to protect the pigs, minimizing the adverse effects caused by the mycotoxin.

Exploring the effect of a microencapsulated citrus essential oil on in vitro fermentation kinetics of pig gut microbiota.

Essential oils (EOs) have emerged as a potential alternative to antibiotics in pig breeding due to their antimicrobial properties. Citrus EOs, a common by-product of the orange juice industry, can be an interesting alternative from a financial perspective due to their huge offer in the global market. Thus, the effect of a citrus EO, and specifically different formulations of Brazilian Orange Terpenes (BOT), on pig gut microbiota was evaluated by means of an in vitro fermentation model simulating different sections of the pig gut (stomach, ileum, and colon). Treatments consisted in: BOT in its unprotected form (BOT, 1.85 and 3.70 mg/mL), microencapsulated BOT (MBOT, 3.50 and 7.00 mg/mL), colistin (2 µg/mL), and a control. BOT and MBOT altered in a similar way the total bacterial 16S rRNA gene copies in the stomach only from 18 h of incubation onwards, and no metabolite production in terms of short-chain fatty acids (SCFAs) was detected. In ileal and colonic fermentations, BOT and MBOT affected ileal and colonic microbiota in terms of total bacterial 16S rRNA gene copies, reduced phylogenetic diversity, and altered composition (p < 0.05) as evidenced by the significant reduction of certain bacterial taxa. However, more pronounced effects were found for MBOT, indicating its higher antimicrobial effects compared to the unprotected BOT, and suggesting that the antibacterial efficiency of the unprotected BOT was probably enhanced by microencapsulation. Furthermore, MBOT stimulated lactate production in ileal fermentations and greatly stimulated overall SCFA production in colonic fermentations. This indicates that besides the shifts in ileal and colonic microbiota by the delivered EO (BOT), the wall material of microcapsules (chitosan/modified starch) might have worked as an additional carbon source with prebiotic functioning, stimulating growth and metabolic activity (SCFAs) of colonic bacteria.

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

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

Essential Oils Extracted from Organic Propolis Residues: An Exploratory Analysis of Their Antibacterial and Antioxidant Properties and Volatile Profile.

The industrial processing of crude propolis generates residues. Essential oils (EOs) from propolis residues could be a potential source of natural bioactive compounds to replace antibiotics and synthetic antioxidants in pig production. In this study, we determined the antibacterial/antioxidant activity of EOs from crude organic propolis (EOP) and from propolis residues, moist residue (EOMR), and dried residue (EODR), and further elucidated their chemical composition. The EOs were extracted by hydrodistillation, and their volatile profile was tentatively identified by GC-MS. All EOs had an antibacterial effect on Escherichia coli and Lactobacillus plantarum as they caused disturbances on the growth kinetics of both bacteria. However, EODR had more selective antibacterial activity, as it caused a higher reduction in the maximal culture density (D) of E. coli (86.7%) than L. plantarum (46.9%). EODR exhibited mild antioxidant activity, whereas EOMR showed the highest antioxidant activity (ABTS = 0.90 µmol TE/mg, FRAP = 463.97 µmol Fe2+/mg) and phenolic content (58.41 mg GAE/g). Each EO had a different chemical composition, but α-pinene and ß-pinene were the major compounds detected in the samples. Interestingly, specific minor compounds were detected in a higher relative amount in EOMR and EODR as compared to EOP. Therefore, these minor compounds are most likely responsible for the biological properties of EODR and EOMR. Collectively, our findings suggest that the EOs from propolis residues could be resourcefully used as natural antibacterial/antioxidant additives in pig production.

Chemical Composition and Antibacterial and Antioxidant Activity of a Citrus Essential Oil and Its Fractions.

Essential oils (EOs) from Citrus are the main by-product of Citrus-processing industries. In addition to food/beverage and cosmetic applications, citrus EOs could also potentially be used as an alternative to antibiotics in food-producing animals. A commercial citrus EO-Brazilian Orange Terpenes (BOT)-was fractionated by vacuum fractional distillation to separate BOT into various fractions: F1, F2, F3, and F4. Next, the chemical composition and biological activities of BOT and its fractions were characterized. Results showed the three first fractions had a high relative amount of limonene (≥10.86), even higher than the whole BOT. Conversely, F4 presented a larger relative amount of BOT's minor compounds (carvone, cis-carveol, trans-carveol, cis-p-Mentha-2,8-dien-1-ol, and trans-p-Mentha-2,8-dien-1-ol) and a very low relative amount of limonene (0.08-0.13). Antibacterial activity results showed F4 was the only fraction exhibiting this activity, which was selective and higher activity on a pathogenic bacterium (E. coli) than on a beneficial bacterium (Lactobacillus sp.). However, F4 activity was lower than BOT. Similarly, F4 displayed the highest antioxidant activity among fractions (equivalent to BOT). These results indicated that probably those minor compounds that detected in F4 would be more involved in conferring the biological activities for this fraction and consequently for the whole BOT, instead of the major compound, limonene, playing this role exclusively.

Laying hens fed mycotoxin-contaminated feed produced by Fusarium fungi (T-2 toxin and fumonisin B1) and Saccharomyces cerevisiae lysate: Impacts on poultry health, productive efficiency, and egg quality.

Mycotoxins represent substantial challenges to the farming industry. These include toxins produced by Fusarium fungi, particularly trichothecenes (toxin T-2) and fumonisin (FB1). In the present study, we determined the effects of addition on Saccharomyces cerevisiae lysate (SCL) added to feed contaminated with T-2 and FB1 in terms of health, productive efficiency, and egg laying quality. We used 60 Hy-line Brown laying hens, and divided them into five groups with four repetitions per group and three birds per repetition. There was one group with no contamination with toxin (NoC). The four other groups included combinations of mycotoxin (4 ppm T-2, and 20 ppm FB1): A contamination group was used as control (the C+ group), and another two contained 500 g/ton of SCL (Detoxa Plus®) (the C + D500 group) or 1000 g/ton of SCL (the C + D1000 group). Finally, one group received feed containing 500 g/ton of Detoxa Plus® and 1000 g/ton of Uniwall® MOS 25 (the C + D500 + U1000 group). The experimental period was 84 days, divided into three productive cycles of 28 days each. The NoC hens had greater egg production than the other groups. Hens that consumed feed with SCL had greater egg production than did the C+ group. The NoC hens produced eggs with greater weights than did the C hens; however, C + D1000 and C + D500 + U1000 birds produced greater egg weights than did the C+ group. The C+ group produced lower egg masses than did the NoC and C + D500 + U1000 groups. The feed intake (FI) was lower in hens that ingested mycotoxin. The use of SCL in feed minimized the negative effects of mycotoxin on feed conversion ratio (FI/dozen). Effects of treatment were detected for feed conversion ratio (kg/kg). The hens that consumed mycotoxin had lower shell resistance and thickness compared to those in the NoC group. The red color of egg yolk was greater in the control groups. There were fluctuations in levels of liver enzymes when birds consumed mycotoxin (sometimes reduced and sometimes increased); nevertheless, the cumulative effect increased the activity of alanine aminotransferase. The serum concentration of reactive oxygen species was greater in hens that ingested mycotoxin only on d 84 compared to the NoC group. Serum glutathione S-transferase activity was greater on d 56 in C + D500 and C + D1000 hens than in the others. We conclude that, in general, the consumption of mycotoxin impaired the performance and quality of the eggs of the hens; the addition of the S. cerevisiae lysate and the addition organic acids, yeast cell wall and mineral carrier minimized some of the negative effects caused by T-2 and FB1.

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

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

Lactobacillus spp. reduces morphological changes and oxidative stress induced by deoxynivalenol on the intestine and liver of broilers.

The mycotoxin deoxynivalenol (DON) contaminates animal feed worldwide, frequently resulting in poor performance and economic losses. Data concerning the effects on poultry health or focusing on intestinal toxicity or the response to oxidative stress are scarce. Also, there is a need for strategies to mitigate the negative effects of DON. This study aimed to investigate the effects of Lactobacillus spp. treatments on the intestine, liver and kidney of poultry fed a DON-contaminated diet. To achieve this aim, histological, morphometrical and histochemical assays were performed. The oxidative stress response was also analyzed by the tests: reduced glutathione, ferric reducing ability, reducing of 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid), nitro blue tetrazolium detection of superoxide anion, and thiobarbituric acid reactive substances. One-day-old broilers chickens (n 50) were submitted to the following treatments: control, DON (19.3 mg kg-1), viable Lactobacillus spp. + DON (VL + DON), heat-inactivated Lactobacillus spp. + DON (HIL + DON), Lactobacillus spp. culture supernatant + DON (LCS + DON). The animals received the contaminated diet for seven days. DON increased the intestinal and liver lesion score, while the Lactobacillus spp. treatments (LT) remained like the control. DON reduced the villi height and increased the crypt depths. The LT showed crypt depths similar to control, and higher villi: crypt ratio in duodenum and jejunum. In the ileum, the LT reduced the goblet cell count in relation to DON group. DON increased the number of intraepithelial lymphocytes (IEL) in jejunum and ileum, while the VL + DON treatment induced a significant decrease in IEL in comparison to DON. DON-diet induced an oxidative stress response in the intestine and liver, and also reduced the antioxidant capacity in these tissues, while LT treatments remained mostly similar to control. DON induced no change in redox balance in the kidney. The LT improved the intestinal health after DON acute exposure, reducing the oxidative stress damage mainly on jejunum and liver.

Pathogenetic effects of feed intake containing of fumonisin (Fusarium verticillioides) in early broiler chicks and consequences on weight gain.

Fusarium verticillioides is often responsible for contamination of poultry feed with the mycotoxin fumonisin. The objective of the study was to determine whether fumonisin-contaminated feed in the early phase of broiler chicks causes oxidative imbalances and interferes with weight gain. One-day-old male Cobb 500 broiler chicks (n = 80) were divided into four treatments of 20 birds each, all of which were fed basal feed until the 11th day of age. From day 12, some birds were challenged with fumonisin in the feed: Control (T0) continued receiving the basal ration; treatments T1, T2, and T3 were given feed experimentally contaminated with fumonisin at concentrations of 2.5 ppm, 5 ppm and 10 ppm, respectively. After the 5th (day 17) and 10th (day 21) days, ten birds from each treatment were euthanized for blood and tissue collection to measure histopathological, biochemical and oxidative stress markers. All animals were weighed individually at the beginning of the experiment (day 12), and at 17 and 21 days of age. Birds that ingested 10 ppm of fumonisin (T3) had lower (P < 0.05) weight gain compared to those in T0. At 21 days, the body weights of the T1, T2 and T3 chicks were 1.3%, 8.97% and 18.7% lower, respectively, than those of T0. No histological lesions in the livers were observed for any treatment; however, higher levels of reactive oxygen species (ROS: day 21) and lipoperoxidation (LPO: days 17 and 21) were observed, associated with lower liver activity of the enzymes superoxide dismutase (SOD: day 21), glutathione peroxidase (GPx: day 17 and 21) and glutathione S-transferase (GST: day 21) when birds consumed 5 or 10 ppm of fumonisin. In serum, LPO levels and SOD and GPx activities were lower for groups consuming high doses of fumonisin in the diet (T2 and T3); ROS levels and GST activity were higher in these birds. Birds that consumed fumonisin-containing diets had lower levels of alanine aminotransferase, total protein and albumin (T3); as well as lower serum glucose levels (days 17 and 21), uric acid and triglycerides (day 21) in T3 than in T0. At 21 days, there were smaller crypt sizes and intestinal villi in birds that consumed high levels of fumonisin. These results suggest that fumonisin (10 ppm) in chick diet causes hepatic oxidative stress and impairs intestinal health, consequently negatively affecting weight gain.

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

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

Fumonisin-(Fusarium verticillioides)-contaminated feed causes hepatic oxidative stress and negatively affects broiler performance in the early stage: Does supplementation with açai flour residues (Euterpe oleracea) minimize these problems?

Fusarium verticillioides is often responsible for contamination of poultry feed with the mycotoxin fumonisin. The aim of this study was to determine whether oxidative stress caused by intake of fumonisin-contaminated feed affects broiler performance at an early stage of development, as well as to test whether the addition of açai residue flour to contaminated feed would minimize these negative effects of redox metabolism. Birds were divided into four groups, with four repetitions of five animals each: control (TC) - birds that received basal feed; TCA treatment - basal feed supplemented with 2% açai flour; TF treatment - feed experimentally contaminated with fumonisin (10 ppm); TFA treatment - fumonisin-contaminated feed (10 ppm) and supplemented with açai fluor (2%). The experiment lasted 20 days, that is, the first 20 days of the chicks' lives. At the end of the experiment, the birds were weighed, and blood, intestine and liver samples were collected. The TCA and TFA had greater body weights and weight gain than did TF. Further, TCA and TFA had lower feed conversion than did TF. Açai flour intake (TCA and TFA) stimulated albumin synthesis and reduced serum AST activity. Nitrate/nitrite (NOx) levels were higher in serum of fumonisin-challenged (TF) birds than in groups; NOx levels were also higher in the livers of all test groups (TF, TCA and TFA) than in TC. Serum glutathione S-transferase (GST) activity was lower in fumonisin-consuming groups (TF and TFA); this was different from what occurred in the liver, that is, higher GST activity in TF and lower activity in TFA than in TC. Catalase activity (CAT) was also higher in the fumonisin-challenged groups (TF and TFA) and the groups supplemented with açai flour (TCA) than in TC. Serum reactive species (RS) and TBARS (lipid peroxidation) levels in the liver were lower in birds supplemented with açai flour and exposed to fumonisin. These data suggest that the addition of açai flour in the feed of early chickens improves animal performance and minimizes the effects of hepatic oxidative stress in birds fed fumonisin-contaminated feed.

Unraveling the selective antibacterial activity and chemical composition of citrus essential oils.

Post-weaning diarrhea (PWD) is an often disease affecting piglets. It is caused mainly by enterotoxigenic Escherichia coli (ETEC) colonization in pig gut. Antibiotics has been used to prevent, combat and control PWD and its negative impact on the productivity of pig breeding sector. Nonetheless, antibiotics due to their wide antibacterial spectrum also can reach beneficial gut bacteria, such as Lactobacillus. Lately, essential oils (EOs) have emerged as a potential alternative to using antibiotics in animal breeding because of their effect on bacterial growth. Commonly, citrus EOs are by-products of food industry and the availability of these EOs in the worldwide market is huge. Thus, six commercials citrus EOs were evaluated on ETEC strains, as model of pathogenic bacteria, and on Lactobacillus species, as models of beneficial bacteria. In overall, citrus EOs exhibited a selective antibacterial activity with higher effect on pathogenic bacteria (ETECs) than beneficial bacteria (Lactobacillus). Brazilian orange terpenes (BOT) oil presented the highest selective performance and caused higher disturbances on the normal growth kinetic of ETEC than on Lactobacillus rhamnosus. The action was dose-dependent on the maximal culture density (A) and the lag phase duration (λ) of the ETEC. The highest sub-inhibitory concentration (0.925 mg/mL) extended the λ duration to ETEC eight times (14.6 h) and reduced A in 55.9%. For L. rhamnosus, the λ duration was only extended 1.6 times. Despite the fact that limonene was detected as the major compound, the selective antibacterial activity of the citrus EOs could not be exclusively attributed to limonene since the presence of minor compounds could be implicated in conferring this feature.

Effects of fed mycotoxin contaminated diets supplemented with spray-dried porcine plasma on cholinergic response and behavior in piglets.

The aim of this study was to evaluate the effect of spray-dried porcine plasma (SDPP) supplementation on cholinesterase enzymes and its relationship with animal behavior of weaning piglets exposed to mycotoxin contaminated diets. To achieve these objectives, two experimental design approaches were used. Male piglets (7.15±0.61kg) 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 desired contamination of 210 µg/kg aflatoxins and 6.690 µg/kg fumonisins; group MYC+SDPP received 253 µg/kg aflatoxins, 6930 µg/kg fumonisins and 6% SDPP. The animals treated with mycotoxin co-contaminated diets showed an increase in AChE and BChE activities in peripheral system (MYC) when compared to control (CTL). Furthermore, supplementation with SDPP (MYC+SDPP group) prevented the mycotoxin-related reduction of AChE in blood and brain. Behavioral tests showed that sleeping and resting behaviors were more often observed in the MYC group; this group also fed fewer times when compared to the other groups, characterizing the deleterious effect of mycotoxins. Taken together, the data suggest changes in AChE and BChE activities may indicate alterations in cholinergic neurotransmission and consequently in the behavior of piglets.

Melaleuca alternifolia essential oil abrogates hepatic oxidative damage in silver catfish (Rhamdia quelen) fed with an aflatoxin-contaminated diet.

Mycotoxins are secondary metabolites produced by varieties of fungi that contaminate food and feed resources and are capable of inducing a wide range of toxicity. This problem is extensively aggravated due to the increasing replacement of fish meal by plant-derived proteins. Among the mycotoxins, aflatoxins have received a great deal of attention owing to their great prevalence in plant feedstuffs and to the detrimental effects on animals. The objective of this study was to evaluate whether dietary supplementation with tea tree (Melaleuca alternifolia) oil (TTO) would avoid or minimize the negative impacts on silver catfish (Rhamdia quelen) fed with aflatoxins-contaminated diets. Four treatments were tested: control (fish fed with a control diet); AFB (fish fed with a mycotoxin-contaminated diet - 1893 µg kg-1 of AFB1 and 52.2 µg kg-1 AFB2); TTO (fish fed with a control diet + 1 mL kg-1 of TTO), and TTO + AFB (fish fed with a mycotoxin contaminated diet - 2324 µg kg-1 of AFB1 and 43.5 µg kg-1 AFB2 + 1 mL kg-1 of TTO). Diets were tested in three replications and analyzed at days 5 and 10 of dietary intake. Significantly reduced antioxidant enzymes (SOD, GPx, and GST) and increased lipid peroxidation (LOOH) and protein carbonyl (PC) content in plasma and liver, with 16.6% mortality occurrence, were observed in the group fed aflatoxin-contaminated diet. Furthermore, aflatoxins also significantly increased plasmatic and hepatic ROS levels and decreased hepatic antioxidant capacity against peroxyl radical (ACAP) levels. Plasma cortisol levels were not altered by aflatoxicosis, but the intoxication induced hepatose. Notwithstanding, addition of TTO to the groups receiving aflatoxins showed a protective effect, avoiding the increase of ROS, LOOH, and PC levels in plasma and liver. Moreover, TTO treatment ameliorated the aflatoxin-associated liver damage. Thus, TTO supplementation at concentration of 1 mL kg-1 in feed may be used in fish to increase antioxidant status and reduce the negative effects caused by aflatoxins toxicity.

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.

Creatine kinase and ATPase activities in piglets fed a fungal mycotoxin co-contaminated diet: Consequences in the pathogenesis of subclinical intoxication.

Creatine kinase (CK) activity, through the creatine-kinase-phosphocreatine (CK/PCr) system, provides a temporal and spatial energy buffer to maintain cellular energetic homeostasis, being responsible to provide adenosine triphosphate (ATP) to the proper function of ATPases enzymes, such as the sodium-potassium (Na+, K+-ATPase) and hydrogen (H+-ATPase) pumps. Thus, the aim of this study was to evaluate the involvement of CK/PCr system in the impairment of energetic homeostasis in piglets fed with a diet co-contaminated with mycotoxins, as well as the effects on ATPases enzymes. Animals were randomly divided in two groups (eight repetitions with two animals each): CON (basal diet) and MYC (mycotoxin diet; 9300 µg/kg of aflatoxins and 8000 µg/kg of fumonisins) which were feed during 15 days. Piglets that received a diet containing 300 µg/kg of aflatoxins and 8000 µg/kg of fumonisins (MYC group) presented lower body weight on days 10 and 15 of experiment when compared to control (CON group). Serum CK activity was lower on days 5, 10 and 15 of experiment in the MYC group. The same occurred for serum Na+, K+-ATPase and H+-ATPase activities on days 10 and 15 when compared to CON group. Moreover, serum calcium levels were superior on day 15 of experiment in the MYC group, while serum potassium and sodium levels were lower in comparison to CON group. Based on these evidences, a diet co-contaminated by aflatoxins and fumonisins inhibits serum CK activity, impairing the energetic homeostasis. This inhibition alters the activities of ATPases (Na+, K+-ATPase and H+-ATPase), contributing to the imbalance of Na+, K+ and Ca+ ionic levels. In summary, the cascade of alterations contributes directly to disease pathogenesis of piglets intoxicated by mycotoxins.

Evaluation of the selective antibacterial activity of Eucalyptus globulus and Pimenta pseudocaryophyllus essential oils individually and in combination on Enterococcus faecalis and Lactobacillus rhamnosus.

Essential oils (EOs), as substitutes for antibiotics in animal diets, should have selective antibacterial activity between pathogenic and beneficial bacteria from the animal gut. Thus, this study evaluated the selective antibacterial activity of Eucalyptus globulus (EG) and Pimenta pseudocaryophyllus (PP) EOs on Enterococcus faecalis as a surrogate model of pathogenic bacterium and on Lactobacillus rhamnosus as a beneficial bacterium model. The EOs antibacterial activity was evaluated by determination of minimal inhibitory concentrations (MICs), minimal bactericidal concentration (MBCs), and fractional inhibitory concentration (FIC) indices. The time-kill and sequential exposure assays were also performed, but using only the EG oil, which was the best selective EO, since it had a MIC lower on E. faecalis (7.4 mg/mL) than on L. rhamnosus (14.8 mg/mL). FIC index values showed that the combination of the two EOs had an indifferent effect (1.25 and 2.03) on E. faecalis and an additive effect (1.00) on L. rhamnosus. The time-kill assay showed that EG oil was able to kill E. faecalis within 15 min of treatment (∼5 log reduction) and caused a reduction ∼3 log of L. rhamnosus viability. The sequential exposure assay showed that EG oil (at MIC/2) produced higher reduction on E. faecalis viability (∼3 log) than on L. rhamnosus (∼2 log) as well. Therefore, L. rhamnosus presented higher tolerance to the antibacterial activity of EG oil than E. faecalis did.

Changes of adenosinergic system in piglets fed a diet co-contaminated by mycotoxin and their effects on the regulation of adenosine.

The effects of diets co-contaminated with 300 µg/kg of aflatoxins and 8000 µg/kg of fumonisins on adenosinergic system of the pigs weaned at 15 days of age were studied. Piglets were inspected daily, and body weight measurement and blood collections were performed at every five days. Piglets intoxicated by mycotoxins presented lower weight gain (p < 0.001) in comparison to control. Intoxicated piglets also showed a reduction in the serum levels of zinc and adenosine and in adenosine deaminase (ADA) activity (p < 0.001). Positive correlations between zinc levels and ADA activity (p < 0.001) and between adenosine levels and ADA activity (p < 0.05) were observed. Ternary plot shows the influence of zinc levels on ADA activity and on adenosine levels, suggesting that low zinc levels, caused by subclinical mycotoxin intoxication, can cause immunomodulatory effects in piglets. We conclude that piglets intoxicated by fumonisins and aflatoxins have low ADA activity and adenosine levels in serum. This can be directly related to zinc reduction, which is a cofactor for ADA. The co-contamination by these mycotoxins in piglet feed impairs growth and immune defenses of the animals, adversely affecting animal health and production. Therefore, changes in the purinergic pathway may affect the pathogenesis of the disease.

Effects of supplementation with spray-dried porcine plasma on blood variables on piglets feed with diet contaminated by mycotoxins.

The aim of this study was to evaluate the supplementation spray-dried porcine plasma (SDPP) effects on body weight, glucose levels and hematological and immunology variables for piglets fed with diet contaminated with low or high aflatoxin and fumonisin levels. Thus, 24 piglets were allotted in four groups, and the group A received a control diet; the group B received a diet with 6% of SDPP; the group C 300 µg/kg of aflatoxins and 8000 µg/kg of fumonisin; the group D 300 µg/kg of aflatoxins, 8000 µg/kg of fumonisin and 6% of SDPP, respectively. A significant increase on hemoglobin concentration and hematocrit levels was observed between C and A groups, result similar also in group D to hemoglobin concentration. A significant reduction on leukocytes and monocytes levels was observed in the group C compared to group A. Most important finding was that SDPP prevented reduction on leukocytes, but not in the monocytes levels in animals of group D. Moreover, the animals of group C presented lower weight, in the same way presented lower on glucose levels. In the other hand, animals in the group C had higher levels of nitrate/nitrite (NOx) and C-reactive protein (CRP) compared other groups. Based on these evidences, the present study showed ingestion of diet contaminated with elevated aflatoxins and fumonisins levels may negatively affected the white cell count and the weight, increasing the seric biomarkers related to inflammatory response, and consequently impairs the immune system. Additionally, SDPP supplementation for piglets prevents the reduction on leukocytes levels and the negative effects associated with weight gain, as well as minimizing the inflammatory response, demonstrating that SDPP can modulate the inflammatory processes and consequently may improve the immune system due active proteins presents in SDPP.