<Emphasis Type="Italic">Fusarium</Emphasis> toxin-contaminated maize in diets of growing bulls: effects on performance,slaughtering characteristics,and transfer into physiological liquids

The present feeding study was carried out to examine the effects of Fusarium toxin-contaminated diets on performance and slaughtering characteristics and on the transfer of the Fusarium toxins zearalenone (ZEN), deoxynivalenol (DON) and their metabolites into physiological matrices. A total of 61 bulls (483?±?46 kg) were fed with graded proportions of Fusarium toxin-contaminated feed over a period of 10 weeks. The total mixed rations (TMR) consisted of 47 % grass silage, 20 % press pulp silage, and 33 % concentrate on dry matter (DM) basis. Increasing toxin concentrations were achieved by the exchange of control maize with Fusarium toxin-contaminated maize in the concentrates. Thus, dietary toxin concentrations between 0.08 and 0.69 mg ZEN and 0.36 and 8.31 mg DON per kg DM were covered by the four feeding groups. Based on increasing DM intake with increasing mycotoxin contaminations of the diet, the live weight gain and energy intake differed significantly between the groups. No effects were observed on slaughtering characteristics and organ weights. ZEN, α-zeralenol, β-zeralenol (β-ZEL), zeralanone, α-zearalanol, β-zearalanol, DON, and de-deepoxy-DON (de-DON) were simultaneously determined in urine, plasma, and liquor whereby quantifiable concentrations of ZEN, β-ZEL, DON, and de-DON were found in urine, of DON and de-DON in plasma, and solely of de-DON in liquor. Based on overall results it can be concluded that current EU-guidance values for critical concentrations of DON and ZEN can be regarded as safe levels also for growing bulls. Urine and blood toxin residue levels can be used to assess exposure of bulls.     

Interactions of zearalenone and its reduced metabolites α-zearalenol and β-zearalenol with serum albumins: species differences,binding sites,and thermodynamics

Zearalenone (ZEN) is a mycotoxin produced by Fusarium species. ZEN mainly appears in cereals and related foodstuffs, causing reproductive disorders in animals, due to its xenoestrogenic effects. The main reduced metabolites of ZEN are α-zearalenol (α-ZEL) and β-zearalenol (β-ZEL). Similarly to ZEN, ZELs can also activate estrogen receptors; moreover, α-ZEL is the most potent endocrine disruptor among these three compounds. Serum albumin is the most abundant plasma protein in the circulation; it affects the tissue distribution and elimination of several drugs and xenobiotics. Although ZEN binds to albumin with high affinity, albumin-binding of α-ZEL and β-ZEL has not been investigated. In this study, the complex formation of ZEN, α-ZEL, and β-ZEL with human (HSA), bovine (BSA), porcine (PSA), and rat serum albumins (RSA) was investigated by fluorescence spectroscopy, affinity chromatography, thermodynamic studies, and molecular modeling. Our main observations are as follows: (1) ZEN binds with higher affinity to albumins than α-ZEL and β-ZEL. (2) The low binding affinity of β-ZEL toward albumin may result from its different binding position or binding site. (3) The binding constants of the mycotoxin-albumin complexes significantly vary with the species. (4) From the thermodynamic point of view, the formation of ZEN-HSA and ZEN-RSA complexes are similar, while the formation of ZEN-BSA and ZEN-PSA complexes are markedly different. These results suggest that the toxicological relevance of ZEN-albumin and ZEL-albumin interactions may also be species-dependent.     

On the distribution and metabolism of Fusarium-toxins along the gastrointestinal tract of endotoxaemic pigs

The aim of this study was to investigate the potential modulatory effect of E. coli lipopolysaccharides (LPS) on residues of deoxynivalenol (DON), de-epoxy-deoxynivalenol (DOM-1), zearalenone (ZEN) and its metabolites α-zearalenol (α-ZEL), β-zearalenol (β-ZEL), zearalanone (ZAN), α-zearalanol (α-ZAL) and β-zearalanol (β-ZAL) after pre- or post-hepatic administration along the gastrointestinal axis. Fifteen barrows were exposed to a naturally mycotoxin contaminated diet (4.59 mg DON/kg feed and 0.22 mg ZEN/kg feed) and equipped with jugular (ju) and portal (po) catheters. On sampling day (day 29), the barrows were infused with LPS or a control fluid (LPS, 7.5 µg/kg body weight; control, 0.9% NaCl) either pre- or post-hepatically, resulting in three infusion groups: CON_ju-CON_po, CON_ju-LPS_po and LPS_ju-CON_po. At 195 min relative to infusion start (210 min post-feeding), pigs were sacrificed and content of stomach and small intestine (proximal, medial and distal part) as well as faeces were collected. In all LPS-infused animals, higher amounts of dry matter were recovered irrespective of LPS entry site suggesting a reduced gastric emptying and a decreased gastrointestinal motility under endotoxaemic conditions. DON metabolism in the gastrointestinal tract (GIT) remained unaltered by treatments and included an increase in the proportion of DOM-1 along the GIT, particularly from distal small intestine to faeces. Variables describing ZEN metabolism suggest a stimulated biliary release of ZEN and its metabolites in LPS-infused groups, particularly in the LPS_ju-CON_po group. In conclusion, the GIT metabolism of ZEN was markedly influenced in endotoxaemic pigs whereby a jugular induction of an acute phase reaction was more effective than portal LPS infusion hinting at a strong hepatic first-pass effect.     

Influence of organically or conventionally produced wheat on health,performance and mycotoxin residues in tissues and bile of growing pigs

From 1999?–?2001 three different varieties of wheat [Contur (susceptible to Fusarium), Batis and Petrus (less susceptible to Fusarium)] were cultivated under organic and conventional conditions in order to determine mycotoxin burden. Soil quality, preceding crop and weather conditions were comparable in the different production systems. The wheat batches were analysed for moulds, and the contents of zearalenone (ZEN) and deoxynivalenol (DON). Feeding trials were carried out with growing pigs (n?=?96; average initial live weight 22.2 ±?1.5?kg [mean?±?SD]) to examine a possible influence on the animal performance and on mycotoxin residues. The data recorded were clinical conditions, performance, biochemical and hematological data. Residues of ZEN, α- and β-zearalenol (ZEL) and of DON were determined in bile, liver and muscle after slaughtering. Conventionally cultivated wheat was more frequently contaminated with Fusarium and contained more frequently ZEN and DON in higher concentrations than the organically produced wheat. Hematological and biochemical parameters of pigs fed with organically cultivated diets were not different from those of conventionally fed pigs. Pigs fed with organically produced wheat showed a slightly higher daily weight gain, but a lower carcass yield than the conventionally fed animals. The highest residues of DON and total-ZEN (ZEN + α-ZEL + β-ZEL) were found in bile. Bile samples of organically fed pigs contained lower concentrations of total-ZEN than those of conventionally fed pigs. Altogether, these data suggest that wheat from an organic farming does not have higher mycotoxin-contamination than wheat from the conventional farming system.     

Urinary deoxynivalenol (DON) and zearalenone (ZEA) as biomarkers of DON and ZEA exposure of pigs

Four diets contaminated with 1.1 to 5.0 mg/kg deoxynivalenol (DON) and 0.4 to 2.4 mg/kg zearalenone (ZEA) were fed to four groups of six growing Large White pigs. Urine samples were collected after 3 to 4 days and again after 6 to 7 days on the diets. On each sampling day, half of the animals were sampled in the morning, after an 8-h fast, and the other half were sampled in the afternoon, after 7 h of ad libitum access to feed. The urinary concentrations of DON, DON-glucuronide, DON-3-sulphate, de-epoxy-DON, as well as of ZEA, ZEA-14-glucuronide, α-zearalenol and α-zearalenol-14-glucuronide, analysed using LC-MS/MS, were used to calculate urinary DON and ZEA equivalent concentrations (DONe and ZEAe). The urinary concentration of DONe (P?<?0.001), but not of ZEAe (P?=?0.31), was lower in the fasted than that in the fed animals. The urinary DONe/creatinine and ZEAe/creatinine ratios were highly correlated with DON and ZEA intake per kg body weight the day preceding sampling (r?=?0.76 and 0.77; P?<?0.001). The correlations between DON intake during the 7 h preceding urine sampling in the afternoon and urinary DONe/creatinine ratio (r?=?0.88) as well as between mean ZEA intake during 3 days preceding urine sampling and urinary ZEAe/creatinine ratio (r?=?0.84) were even higher, reflecting the plasma elimination half-time of several hours for DON and of more than 3 days for ZEA. ZEAe analysed in enzymatically hydrolysed urine using an ELISA kit was highly correlated with the LC-MS/MS data (r?=?0.94). The urinary DONe and ZEAe to creatinine ratios, analysed in pooled urine samples of several pigs fed the same diet, can be used to estimate their exposure to DON and ZEA.     

Occurrence of deoxynivalenol and zearalenone in brewing barley grains from Brazil

Barley (Hordeum vulgare L.) is an important cereal crop for food and represents one of the main ingredients in beer production. Considering the importance of barley and its derived products, the knowledge about the mycotoxin contamination in the barley production is essential in order to assess its safety. In this study, the levels of deoxynivalenol (DON) and zearalenone (ZEN) in brewing barley were determined using a LC-MS/MS method. A survey was conducted in 2015 to estimate the mycotoxin levels in these products (n?=?76) from four crop regions in Brazil. The results showed high levels of DON and ZEN in the analyzed samples, with contamination levels of 94 and 73.6%, respectively. The mean levels of DON and ZEN ranged from 1700 to 7500 μg/kg and from 300 to 630 μg/kg, respectively. Barley samples from regions 1 and 2 presented higher levels of ZEN and DON, respectively, and those from region 4 presented lower levels of both. Co-occurrence of DON and ZEN was seen in the majority of the barley grain samples, and the mycotoxin content was above the maximum levels established by the Brazilian and European regulations.     

Feasibility of 3D UV-C treatment to reduce fungal growth and mycotoxin loads on maize and wheat kernels

Fungal disease of grain crops is a concern for the agricultural industry, resulting in economic losses. Aside from severe yield losses, mycotoxigenic fungi such as Penicillium and Fusarium can produce harmful mycotoxins, including deoxynivalenol (DON), zearalenone (ZEN), and ochratoxin A (OTA). This proof-of-concept study explored the feasibility and effects of ultraviolet (UV) C light at 253.7 nm to reduce fungal and mycotoxin loads on model surfaces as well as on maize and wheat kernels using benchtop 2D and 3D illumination strategies. Reduction of Penicillium verrucosum (98.6%) and Fusarium graminearum (88.8%) on agar was achieved using a UV-C dose of 100 mJ cm^?2. Naturally occurring fungal growth resembling P. verrucosum on maize was reduced by 79% after exposure to 5000 mJ cm^?2. Similarly, fungal growth resembling F. graminearum on maize was reduced by 60% with 1000 mJ cm^?2. On wheat, significant reduction of fungal growth was not observed. Maximal reduction of DON (97.3%), ZEN (75.4%), and OTA (91.2%) on filter paper was obtained using 15,000 mJ cm^?2. The overall reduction of DON (30%; 14%), ZEN (52%; 42%), and OTA (17%; 6%) on maize and wheat, respectively, was lower than on filter paper. Moisture and crude protein content as well as percent germination of maize kernels were not affected by UV-C treatment up to 5000 mJ cm^?2. This study has shown that 3D UV-C treatment is a feasible option for reducing Fusarium and Penicillium growth on maize kernels and, at higher doses, decreasing ZEN by ~?50%.     

An angiotensin-converting enzyme-inhibitory metabolite with partial structure of microginin in a cyanobacterium <Emphasis Type="Italic">Anabaena fertilissima</Emphasis> CCC597, producing fibrinolytic protease

Whole cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) of Anabaena fertilissima CCC597 revealed the presence of a microginin (M _r?=?565 Da), which was purified using ODS C18 solid-phase extraction followed by quaternary methyl ammonium (QMA)-ion exchange and reverse-phase liquid chromatography. Its partial structure was deduced upon LC-MS/MS as β-amino-α-hydroxy-decanoic acid-Ala-MeLeu-Tyr. Microginin-565 inhibited rabbit angiotensin-converting enzyme activity at an apparent IC₅₀?=?9.8 μM.     

Mycotoxins in organic and conventional cereals and cereal products grown and marketed in Croatia

In this study, the levels of aflatoxin B₁ (AFB₁), ochratoxin A (OTA), zearalenone (ZEN), deoxynivalenol (DON) and fumonisins (FUM) in unprocessed cereals (n = 189) and cereal-based products (n = 61) were determined using validated ELISA methods. All samples originated from either conventional or organic production corresponded to the 2015 harvest in Croatia. Based on the mean mycotoxin concentrations, the risk for the consumer to exceed the tolerable daily intake (TDI) for these toxins by the consumption of both types of cereals and cereal-based products was assessed. Mycotoxin contamination of organic cereals and organic cereal-based products was not significantly different (p > 0.05). Given that the exposure assessment resulted in a small fraction of the TDI (maximum: DON, 12% of TDI), the levels of the investigated mycotoxins in both types of cereals and cereal-based products from the 2015 harvest did not pose a human health hazard.     

Modeling population dynamics of yeast-like symbionts (Ascomycota: Pyrenomycetes: Clavicipitaceae) of the planthopper <Emphasis Type="Italic">Delphacodes kuscheli</Emphasis> (Hemiptera: Delphacidae)

Delphacodes kuscheli establish mutualistic relationship with yeast-like symbionts (YLS) that live in the fat body and are necessary for host survival and reproduction. We estimated for a host of age t, its body weight, W_(t), and the number of YLS per host, YLS_(t). The host body weight was calculated as: W_(t)?=?Lm/[1+ e ^(d–kt)], (Lm?=?the maximum observed weight, and d and k are constants), and the fat body was considered a fixed proportion of W_(t). We calculated the number of YLS per unit host body mass: α_(t)?=?YLS_(t)/W_(t). We also calculated the number of YLS per host, cYLS_(t), and analyzed the pattern of variation in both sexes adapting the expression of the logistic model: cYLS_(t)?=?KN_oe^rt/K+(e^rt -1)N_o, (N_o?=?initial number of YLS, r?=?intrinsic per capita rate of natural increase, and K?=?variable carrying capacity). In females the carrying capacity varied according to a constant proportion of the host’s weight: K_(t)?=?αW_(t). In males α_(t) was considered a decreasing function of the host age: K_(t)?=?α_(t)W_(t). The coefficients No, α, and r were subjected to parameterization. We found that the patterns of W_(t) and YLS_(t) of D. kuscheli were similar to other planthoppers. In females YLS increased up to the adult stage and then remained almost constant, varying similarly to individual weight. In males YLS increased up to the 5th instar nymph as the individual weight did, but the number of YLS decreased in the adult stage and the correlation was not so good. The calculated number of YLS per host matches reasonably well with the number estimated experimentally both in females and males. This is the first study that quantified and modeled the dynamics of YLS endosymbionts in a Neotropical planthopper pest. The models will be used in future studies for better understand the experimental reduction of YLS in young nymphal stages.     

Population genetics of <Emphasis Type="Italic">Cedrela fissilis</Emphasis> (Meliaceae) from an ecotone in central Brazil

Cedrela fissilis is an endangered timber species associated with seasonal forests throughout South America. We investigated a population of C. fissilis (PAN) located toward central Brazil to uncover insights on how an ecotone may have shaped the evolutionary history of this species at the local scale. PAN consisted of 18 mother trees and their 283 offspring (18 families), which were genotyped with ten microsatellite loci. We supplemented our dataset with equivalent microsatellite data from 175 specimens representing the east and west lineages of C. fissilis. An array of complementary methods assessed PAN for genetic diversity, population structure, and mating system. In PAN, the gene pool of the east lineage combined with a third (previously unidentified) lineage to form an admixture population. PAN is under inbreeding (Ho?=?0.80 and 0.74, uHe?=?0.85 and 0.82, Ap?=?1.1 and 7.1, F?=?0.06 and 0.10, for mother trees and offspring, respectively). Mother trees were predominantly outcrossing (t_m?=?0.95), with some selfing (1???t_m?=?0.05), and crossing between related individuals (t_m–t_s?=?0.07); they received pollen from few donors (N_ep?=?9). Restricted gene flow within PAN gave rise to a strong population structure, which split the 18 families into six groups. Some mother trees were reproductively isolated. Conservation perspectives are discussed.     

Plasma kinetics and matrix residues of deoxynivalenol (DON) and zearalenone (ZEN) are altered in endotoxaemic pigs independent of LPS entry site

This study aimed to investigate a potential modulatory effect of E. coli lipopolysaccharide (LPS) on the kinetics of deoxynivalenol (DON) and zearalenone (ZEN) after pre- or post-hepatic LPS administration to unravel the putative role of the liver. Fifteen barrows were fed a diet containing mycotoxin-contaminated maize (4.59 mg DON/kg feed, 0.22 mg ZEN/kg feed) for 29 days and equipped with pre-hepatic catheters (portal vein, “po”) and post-hepatic catheters (jugular vein, “ju”), facilitating simultaneous infusion of LPS (“LPS group”, 7.5 μg/kg body weight) or 0.9% sterile NaCl solution (control, “CON group”, equivolumar to LPS group) and blood sampling. This resulted in three infusion groups, depending on infusion site: CON_ju-CON_po, CON_ju-LPS_po, and LPS_ju-CON_po. On day 29, pigs were fed their morning ration (700 g/pig) (?15 min), and blood samples were collected at regular intervals relative to infusion start. At 195 min, pigs were sacrificed and bile, urine, liquor, and liver samples collected. DON concentrations in jugular and portal blood decreased in both LPS-infused groups, whereas the ZEN concentrations increased, regardless of the treatment site. In liver tissue, a decrease of both toxin concentrations was observed in endotoxaemic pigs as well as a drop in hepatic conjugation, regardless of LPS entry site. In contrast to our hypothesis, DON and ZEN were not differently altered depending on the LPS-entry site. Neither the absorption nor the accumulation of DON and ZEN in different tissues differed significantly between animals which were infused with LPS via either the jugular or portal vein.     

High genetic diversity in an endangered medicinal plant, <Emphasis Type="Italic">Saussurea involucrata</Emphasis> (<Emphasis Type="Italic">Saussurea</Emphasis>, Asteraceae), in western Tianshan Mountains,China

Saussurea involucrata (Asteraceae) is a medicinal and second-degree national priority endangered plant that is mainly distributed in the high latitude region of the western Tianshan Mountains. The population is fragmented and isolated, and extensive human impact merits a suitable and specific conservation strategy, which can be compiled based on the genetic diversity, population structure, and demographic history. Phylogeographic studies were conducted on a total of five natural populations and 150 individuals were sampled. Data from three cpDNA intergenic spacer regions (trnL-F, matK, and ndhF-rpl32) and nrDNA ITS sequences showed that twelve haplotypes in cpDNA and five haplotypes in nrDNA indicated high genetic diversity among populations sampled (H _T?=?0.820 and 0.756) and within populations sampled (H _S?=?0.792 and 0.721). Additionally, the high genetic diversity did not mirror genetic structure in either cpDNA (F _ST?=?0.03153, G _ST?>?N _ST, p?<?0.05) or nrDNA (F _ST?=?0.03666, meaningless G _ST and N _ST). Two groups (north and south) were determined for a SAMOVA analysis. Based on this analysis, the demographic history was conducted with a Bayesian Skyline Plot and Isolation with Migration analysis, which showed sustainable and stable extension without a marked bottleneck. Divergence time was indicated at c. 6.25 Mya (90%HPD: 15.30–0.22 Mya) in the Miocene, which is consistent with the formation of the Kelasu section of Tianshan. The southern populations in the Bayanbulak and Gonglu regions require additional attention and transplanting would be an effective way to restore rare cpDNA haplotypes, increase effective population size, and migration rate. Our results suggested that in situ conservation of S. involucrata in western Tianshan should be the main strategy for protection and recovery of the species.     

Effective to census population size ratios in two Near Threatened Mediterranean amphibians: <Emphasis Type="Italic">Pleurodeles waltl</Emphasis> and <Emphasis Type="Italic">Pelobates cultripes</Emphasis>

Efforts to mitigate amphibian declines are hindered by a lack of information about basic aspects of their biology and demography. The effective to census population size ratio (N _e /N _c ) is one of the most important parameters for the management of wildlife populations because it combines information on population abundance and genetic diversity and helps predict population viability in the long term. Few studies have calculated this ratio in amphibians, which sometimes show low ratios, associated with a higher extinction risk. Here we integrate field-based (capture-mark-recapture studies, egg string counts) and molecular approaches (estimation of the effective number of breeders (N _b ) and the effective population size (N _e ) based on genotypes from larval cohorts and candidate parents) to produce the first estimates of the N _e /N _c and N _b /N _c ratios in two amphibians, the Iberian ribbed newt Pleurodeles waltl and the western spadefoot Pelobates cultripes. Additionally, we investigate sex-biased dispersal in both species based on direct (field observations) and indirect (genetic) evidence. Both species showed similar ratios, slightly lower in Pleurodeles (0.21–0.24) than in Pelobates (0.25–0.30). Observed displacement rates were low in both species (P. waltl?=?0.51%; P. cultripes?=?1.23%). We found no evidence for sex-biased dispersal in P. cultripes, but both direct and indirect evidences suggest a tendency for female-biased dispersal in P. waltl. We discuss differences in the genetic estimates of N _e and N _b provided by three inference methods and the implications of our findings for the management of these species, characteristic of Mediterranean wetlands in the Iberian Peninsula and listed as Near Threatened.     

Expressions of some antioxidant genes in SH-SY5Y cells treated with β-lapachone,morphine and electromagnetic field

β-Lapachone (β-Lap), morphine (Mor), and electromagnetic field (EMF) generate reactive oxygen species. The goal of the present study was to examine the effects of Mor and EMF, in combination with β-Lap on the cell growth inhibition and expression of several antioxidant genes. The 0.50 mT intensity of 50 Hz EMF and two exposure conditions (“15 min field-on/15 min field-off” and “30 min field-on continuously”) on SH-SY5Y cells were used. The effects of Mor and EMF, in combination with β-Lap on cell growth inhibition and the expression levels of several antioxidant genes (NQO1, NQO2, SOD1, SOD2, CAT, GSTO1, GSTM2, GSTM3, GSTP1, MGST1, MGST3) in SH-SY5Y cells were measured. The relative mRNA levels were calculated according to the \({{\text{2}}^ - }^{{\Delta \Delta {{\text{C}}_{\text{t}}}}}\). Whereas NQO1 mRNA level decreased in the “15 min field-on/15 min field-off” condition, the expression level of NQO2 was increased. Both NQO1 and NQO2 expressions increased in Mor treated cells. IC₅₀ values of β-Lap in combination with Mor, EMF, and “Mor?+?EMF” were higher than cells treated only with β-Lap. The NQO1 expression level in the cells treated with β-Lap was higher than the other treatments, indicating that β-Lap induces the expression of NQO1. Moreover, multiple linear regression analysis indicated that NQO1 mRNA levels were associated positively with β-Lap and negatively with EMF. At least in part, the mRNA levels of NQO1 were associated with IC₅₀ values of β-Lap in designed treatments. There is a negative association between mRNA levels of NQO1 and IC₅₀ values of β-Lap but not NQO2.     

Evaluation of <Emphasis Type="Italic">Galleria mellonella</Emphasis> larvae as an in vivo model for assessing the relative toxicity of food preservative agents

Larvae of Galleria mellonella are widely used for evaluating the virulence of microbial pathogens and for measuring the efficacy of anti-microbial agents and produce results comparable to those that can be obtained using mammals. In this work, the suitability of using G. mellonella larvae to measure the relative toxicity of a variety of food preservatives was evaluated. The response of larvae to eight commonly used food preservatives (potassium nitrate, potassium nitrite, potassium sorbate, sodium benzoate, sodium nitrate, sodium chloride, sodium nitrite and sodium acetate) administered by feeding or by intra-haemocoel injection was measured. A significant correlation between the LD₅₀ (R ²?=?0.8766, p?=?0.0006) and LD₈₀ (R ²?=?0.7629, p?=?0.0046) values obtained due to oral or intra-haemocoel administration of compounds was established. The response of HEp-2 cells to the food preservatives was determined, and a significant correlation (R ²?=?0.7217, p?=?0.0076) between the LD₅₀ values of the compounds administered by feeding in larvae with the IC₅₀ values of the compounds in HEp-2 cells was established. A strong correlation between the LD₅₀ values of the eight food preservatives in G. mellonella larvae and rats (R ²?=?0.6506, p?=?0.0156) was demonstrated. The results presented here indicate that G. mellonella larvae may be used as a model to evaluate the relative toxicity of food preservatives, and the results show a strong positive correlation to those obtained using established cell culture and mammalian models.     

Effects of deoxynivalenol (DON) and its microbial biotransformation product deepoxy-deoxynivalenol (DOM-1) on a trout,pig, mouse,and human cell line

Deoxynivalenol (DON), a trichothecene produced by various Fusarium species, is one of the most prevalent food- and feed-associated mycotoxins. The effects of DON and deepoxy-deoxynivalenol (DOM-1) were assessed in five different cell lines from different tissues and species starting from the first line of defense, the trout gill (RTgill-W1) and pig intestinal cells (IPEC-1 and IPEC-J2) over immune cells, as second line of defense (mouse macrophages RAW 264.7) to human liver cells (HepG2). Viability was assessed with a WST-1 assay, except for RTgill-W1, where a neutral red (NR) and sulforhodamine B (SRB) assay was performed. Additionally, more sensitive parameters, such as interleukin-, nitric oxide (NO)-, and albumin-release were determined. Viability was affected by DON at concentrations starting at 10 μmol/L (RTgill-W1), 0.9 μmol/L (IPEC-1), 3.5 μmol/L (IPEC-J2), and 0.9 μmol/L (HepG2), whereas DOM-1 did not have such an effect. Additionally, NO was decreased (0.84 μmol/L DON), whereas interleukin (IL)-6 was increased (0.42 μmol/L DON) in lipopolysaccharide (LPS)-stimulated DON-, but not DOM-1-treated RAW cells. Tumor necrosis factor (TNF)-α release, however, was not affected. Interestingly, albumin secretion of HepG2 cells was decreased by both DON and DOM-1 but at a much higher concentration for DOM-1 (228 versus 0.9 μmol/L for DON). 98.9% of DOM-1 was retrieved by liquid chromatography tandem mass spectrometry at the end of the experiment, proving its stability. In this study, IL-6 was the most sensitive parameter, followed by NO and albumin release and viability for HepG2 and IPEC-1.     

The potential effects of antioxidant feed additives in mitigating the adverse effects of corn naturally contaminated with <Emphasis Type="Italic">Fusarium</Emphasis> mycotoxins on antioxidant systems in the intestinal mucosa,plasma, and liver in weaned pigs

Seventy-two piglets (6.0 kg BW) were randomly distributed within six different dietary treatments to evaluate the effect of deoxynivalenol (DON) and the potential of four antioxidant feed additives in mitigating the adverse effects of DON on growth performances and oxidative status. Dietary treatments were as follows: control diet 0.8 mg/kg DON; contaminated diet (DON-contaminated diet) 3.1 mg/kg DON; and four contaminated diets, each supplemented with a different antioxidant feed additive, DON + vitamins, DON + organic selenium (Se)/glutathione (GSH), DON + quercetin, and DON + COMB (vitamins + Se/GSH + quercetin from the other treatments). Although DON was the main mycotoxin in the contaminated diet, this diet also contained 1.8 mg/kg of zearalenone (ZEN). The “mycotoxin” effects therefore included the combined effect of these two mycotoxins, DON, and ZEN. The DON-ZEN ingestion did not affect growth performances, average daily gain (ADG), average daily feed intake (ADFI), and feed efficiency (G:F ratio), but partially induced oxidative stress in weaned pigs as shown by increased malondialdehyde (MDA) content in the plasma and superoxide dismutase (SOD) activity in liver (P?<?0.05). However, no change in the activity of other antioxidant enzymes or GSH concentrations was observed in plasma and liver of piglets fed the DON-contaminated diet (P?>?0.05). Supplementation with individual antioxidant feed additive had a limited effect in weaned pigs fed DON-ZEN-contaminated diets. Combination of antioxidants (vitamins A, C, and E, quercetin, and organic Se/GSH) reduced plasma and liver MDA content and SOD activity in liver (P?<?0.05) of piglets fed DON-ZEN-contaminated diets. Furthermore, this combination also reduced MDA content in the ileum (P?<?0.05), although activity of glutathione peroxidases (GPx), SOD or catalase (CAT) in the ileum was not affected by DON-ZEN contamination or antioxidant supplements. In conclusion, DON-ZEN contamination induced oxidative stress in weaned pigs and combination of antioxidant feed additives restored partially the oxidative status. Further studies will be necessary to assess whether the effects of antioxidant feed additives on oxidative status are specific when feed is contaminated with DON-ZEN.     

Immunochemical Localization of GABA<Subscript>A</Subscript> Receptor Subunits in the Freshwater Polyp <Emphasis Type="Italic">Hydra vulgaris</Emphasis> (Cnidaria,Hydrozoa)

γ-aminobutyric acid (GABA) receptors, responding to GABA positive allosteric modulators, are present in the freshwater polyp Hydra vulgaris (Cnidaria, Hydrozoa), one of the most primitive metazoans to develop a nervous system. We examined the occurrence and distribution of GABA_A receptor subunits in Hydra tissues by western blot and immunohistochemistry. Antibodies against different GABA_A receptor subunits were used in Hydra membrane preparations. Unique protein bands, inhibited by the specific peptide, appeared at 35, 60, ～50 and ～52 kDa in membranes incubated with α3, β1, γ3 or δ antibodies, respectively. Immunohistochemical screening of whole mount Hydra preparations revealed diffuse immunoreactivity to α3, β1 or γ3 antibodies in tentacles, hypostome, and upper part of the gastric region; immunoreactive fibers were also present in the lower peduncle. By contrast, δ antibodies revealed a strong labeling in the lower gastric region and peduncle, as well as in tentacles. Double labeling showed colocalization of α3/β1, α3/γ3 and α3/δ immunoreactivity in granules or cells in tentacles and gastric region. In the peduncle, colocalization of both α3/β1 and α3/γ3 immunoreactivity was found in fibers running horizontally above the foot. These data indicate that specific GABA_A receptor subunits are present and differentially distributed in Hydra body regions. Subunit colocalization suggests that Hydra GABA receptors are heterologous multimers, possibly sub-serving different physiological activities.     

Synthesis and Protective Activity of β-Glycosides of <Emphasis Type="Italic">N</Emphasis>-Acetylmuramyl-<Emphasis Type="Italic">L</Emphasis>-Alanyl-<Emphasis Type="Italic">D</Emphasis>-Isoglutamine with Alkylalicyclic and Arylaliphatic Aglycons

The following glycosides of N-acetylmuramyl-L-alanyl-D-isoglutamine (MDP) were synthesized: β-4-tert-butylcyclohexyl MDP, β-2-(adamant-1-yl)ethyl MDP, β-2,2-diphenylethyl MDP, and β-2-(p-biphenyl) ethyl MDP. The starting peracetylated β-N-acetylglucosaminides were prepared by the oxazoline method. They were converted into 4,6-O-isopropylidene-N-acetyl-D-muramic acids, which were coupled with L-Ala-D-Glu(NH₂)OBn. The target glycopeptides were obtained after their deprotection. The stimulation of the anti-infection resistance of mice against Staphylococcus aureus by the MDP glycosides was studied.