Desensitization of exposed root surfaces using a semilunar coronally positioned flap.

A definitive treatment is described for cervical dentinal sensitivity, featuring a semilunar coronally positioned flap to cover the denuded root surface. Many patients who complain of having sensitive teeth from exposed dentin secondary to gingival recession can be easily and conservatively treated with a semilunar flap procedure.

Effect of the appetite stimulant cyproheptadine on deoxynivalenol-induced reductions in feed consumption and weight gain in the mouse.

Deoxynivalenol (DON, vomitoxin), a Fusarium mycotoxin, is suspected of inducing its anorectic/feed refusal activity through a serotoninergic (5HT) mechanism, possible via 5HT2-receptors. In this study the efficiency of cyproheptadine (CYP), a serotonin antagonist and known appetite stimulant, to attenuate the adverse effect of DON was investigated in mice. CYP was administered in the feed for two days before animals began receiving the DON, which was also added to the feed. Both agents were administered concurrently thereafter for a 12-day period. Dosing levels included various combinations of the two compounds, ranging from 0-16 ppm DON and 0-20 ppm CYP.

The role of tryptophan in DON-induced feed rejection.

Deoxynivalenol (DON, vomitoxin) commonly produced by Fusarium fungi can alter feeding behaviour of pigs and other farm animals. The effects of dietary tryptophan (TRP, precursor of brain amine serotonin) in combination with DON were examined in mice to determine if TRP can modulate DON toxicity. Results indicated that brain TRP can be influenced by dietary TRP, but no evidence of TRP potentiating DON toxicity was observed. Higher TRP levels likely induced amino acid imbalance leading to weight gain suppression.

Response of growing swine to dietary exposure to pure fumonisin B1 during an eight-week period: growth and clinical parameters.

Consumption of corn or corn-based products contaminated with Fusarium moniliforme/fumonisins has been associated with a variety of animal and human diseases and is a major food/feed safety issue. This study focused on the clinical toxicity and performance parameters in growing swing exposed to low to moderate levels of pure fumonisin B1 (FB.) for 8 weeks. Male (castrated) and female pigs were fed diets containing 0,0.1,1.0, and 10 mg FB1/kg diet (ppm). Weight gains and feed consumption were measured weekly. Blood samples were collected throughout the study, and various clinical and hematological parameters were measured. Because fumonisins are potent inhibitors of sphingolipid biosynthesis, sphinganine and sphingosine concentrations were determined in the liver, lung, and kidney. Organ weights and carcass quality were measured at the end of the trial. In general, male pigs were more adversely affected by FB1 in the diet than females. The average daily gain for males decreased by 8% for pigs fed 1.0 ppm and by 11% at 10.0 ppm, when compared to the control (0 ppm). Males fed 0.1 ppm showed an erratic growth pattern during the first 5 weeks of the experiment. Feed consumption for the same animals was somewhat higher than that of the controls during each of the first 4 weeks but thereafter was 6-7% lower each week as compared to controls. Female pigs fed FB1-diets showed a general enhancement of feed consumption until week 4. Among clinical chemistry parameters, cholesterol increased in males for the 1.0 and 10.0 ppm diets as compared to controls after 2 weeks, while the levels in both sexes were elevated for the 1.0 ppm diet only by the end of the experiment. Serum liver enzyme concentrations were altered during week 2 only. Changes were observed in the weight of the pancreas and adrenals for male pigs fed FB1 diets as compared to controls. The free sphinganine to free sphingosine ratio (biomarker of exposure in FB1-consuming animals) increased in all three organs for the 10 ppm diet, regardless of sex. The study indicated that FB1 can cause different effects at each dose level, at concentrations as low as 0.1 ppm (showing erratic growth) followed by a reduced growth and biochemical abnormalities in blood (1.0 ppm) and sphingolipid alterations in tissues (10.0 ppm). Some of these effects occurred below the exposure level that caused alteration in sphingolipid metabolism.

Influence of low-level exposure to Fusarium mycotoxins on selected immunological and hematological parameters in young swine.

The effects of low dietary concentrations of Fusarium mycotoxins (deoxynivalenol (DON), 15-acetyl-DON, and zearalenone) on growth, immunological, and hematological parameters were determined in young pigs during a 28-day feeding experiment. Clean and naturally contaminated corn were incorporated into basal diets formulated to contain 0.00, 0.75, 1.50, and 3.00 mg DON/kg diet. A pair-fed control animal was used for comparison with each animal receiving the highest level of contamination (diet 4). Skin temperature, measured during the first week of the experiment, decreased linearly as the dietary mycotoxin concentration increased. Several other linear effects were observed: depressed feed intake throughout the experiment, reduction in thyroid size (absolute/relative), and changes in the appearance of the esophageal region of the stomach (thicker and higher degree of folding with increasing toxin concentration). Serum T4 (thyroxine) levels increased quadratically after 7 and 28 days of exposure compared to control animals. This change coincided with an increase in albumin levels, a decrease in alpha-globulin levels, and an overall increase in albumin/globulin ratio as the level of contamination increased. After immunization with sheep red blood cells (SRBC), animals fed contaminated diets showed a delayed response in peak titers. At the end of the experiment an increase in the segmented neutrophil count was observed. The following observations were made for animals consuming diet 4 as compared to the pair-fed controls: lower skin temperature, better feed efficiency, more corrugated stomachs, reduced alpha-globulin levels, and lower antibody titers to SRBC.(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of level of deoxynivalenol in the diet of dairy cows on feed intake, milk production, and its composition.

Eighteen primiparous Holstein cows were used in a 10-wk lactation study, preceded by a 2-wk covariate period, to determine the effect of concentration of deoxynivalenol in the diet on cow performance and transfer of deoxynivalenol and its metabolite, deepoxydeoxynivalenol, to milk. Diets were formulated to contain deoxynivalenol at 0, 6, and 12 mg/kg of concentrate DM, and daily intake of deoxynivalenol was .59, 42, and 104 mg, respectively. Increasing deoxynivalenol in the diet did not affect intake of concentrate or forage. Total milk output was not affected; however, milk fat responded quadratically; cows given deoxynivalenol at 6 mg/kg of concentrate DM had the lowest milk fat content and fat output. Overall energetic efficiency was not influenced because reduced energy output in milk was compensated by increased BW gains. No transfer of deoxynivalenol or deepoxydeoxynivalenol to milk was observed; concentrations were below detectable limits (1 microgram/ml) using HPLC-mass spectroscopy. We concluded that diets containing deoxynivalenol up to 6 mg/kg of dietary DM did not reduce feed intake of cows in this study and that deoxynivalenol or deepoxydeoxynivalenol was not transferred to milk. Further studies are required to confirm the apparent lack of effect of deoxynivalenol on milk production.

Rapid colorimetric bioassay for screening of Fusarium mycotoxins.

The cytotoxicity of Fusarium mycotoxins was evaluated using a trichothecene sensitive cell line (BHK-21, baby hamster kidney cells) in combination with the MTT-cleavage test as an end-point measurement. Cells tended to be more sensitive to the type A trichothecenes with midpoint cytotoxicity values ranging from 1.6 ng/ml for T-2 toxin to 60 ng/ml for scirpentriol. The cytotoxicity value for deoxynivalenol (type B) was 112 ng/ml. The inherent disadvantage of the MTT-assay (formation of insoluble formazan) was overcome by using the analog MTS and measuring the water-soluble formazan directly in the culture media. The MTS-midpoint cytotoxicity values for T-2 toxin and deoxynivalenol (2.1 and 141 ng/ml, respectively), although slightly higher, showed a good correspondence to the MTT-test. Both the MTT- and MTS-cleavage tests are useful for evaluating the cytotoxicity of Fusarium mycotoxins. The replacement of MTT by MTS substantially reduced the number of sample processing steps and the length of time required to complete the cytotoxicity assay.

Investigations in the use of mice exposed to mycotoxins as a model for growing pigs.

A series of experiments was conducted to determine the feasibility of using mice to screen for possible dietary mycotoxin interactions before testing them with swine. Selected mycotoxins, deoxynivalenol (DON) and T-2 toxin, were fed to young mice, alone and in combination. The severity of effects on body weights caused by DON (0-20 mg DON/kg diet) was more pronounced in a dose-related manner when the animals were exposed to contaminated diets starting at 21 d of age than at 28 d (Experiment 1) as reflected in the analysis of variance. The relative variance among diets after 7 d was twice as great for the younger than for the older mice. In both age groups, the weight gain response was linear, similar to that seen in growing swine. In Experiment 2, a significant (p < .05) diet type x DON interaction for food consumption evident after 7 d, indicated that the effect of DON depended on the type of diet (freeze-dried vs. regular mash). There was no difference in food efficiency between diet type, but a strong dose-dependent effect due to DON was observed. When DON and T-2 toxin were fed together to young mice, a significant (p < .001) linear decrease in weight gain and food consumption was observed after 7 d on the contaminated diet as the toxin concentration increased.

Effect of deoxynivalenol on neurotransmitters in discrete regions of swine brain.

The effect of deoxynivalenol (DON, vomitoxin) on brain amine levels was investigated in swine. DON, a trichothecene mycotoxin, causes suppression of feed intake (anorexia) in susceptible species. Following acute administration of DON to pigs (0.25 mg/kg, IV), concentrations of endogenous catecholamines norepinephrine (NE), dopamine (DA), 3,4-dihydroxyphenyl-acetic acid (DOPAC), and homovanillic acid (HVA), and the indoleamines, 5-hydroxytryptamine (5HT, serotonin) and 5-hydroxyindoleacetic acid (5HIAA) were determined in five brain regions, periodically during the 24 h post-dosing. Analysis was carried out by high performance liquid chromatography, using electrochemical detection. Effects of DON in the swine brain were transmitter, time and region-specific. It was observed that levels of the major transmitters (NE, DA and 5HT) were statistically different from controls in the hypothalamus (Hypo), frontal cortex (FCX) and cerebellum (Cb) up to 8 h post-dosing. Overall, DON administration elevated NE and depressed DA concentrations in these regions, and levels of 5HT which increased initially in Hypo (1 h), had dropped significantly below controls in both Hypo and FCX at 8 h. These alterations, however, were not indicative of known neurochemical changes associated with chemical-induced anorexia. Instead, this data suggested that the neurochemical effects of acute DON exposure might be due to peripheral toxicological events (i.e., vomiting), which overwhelmed its more subtle feed refusal activity.

Evaluation of potential interactions involving trichothecene mycotoxins using the chick embryotoxicity bioassay.

The Chick Embryotoxicity Screening Test (CHEST) was used to examine possible interactions between different trichothecenes. Combinations of deoxynivalenol (DON), 15-acetyldeoxynivalenol (15-ADON) and HT-2 toxin were tested at various concentrations and the effects were determined based on percent mortality. The combined toxicity of any two trichothecenes was found to be additive. Discrepancies between observed and expected mortality values seldom differed by more than 10%, well within sampling error for the binomial distribution. Several limitations of the CHEST assay are discussed.

Testing a short-term feeding trial to assess compositional and histopathological changes in hearts of rats fed vegetable oils.

Male, female and castrated rats, three wk of age, were fed a low-fat diet for 14 wk followed by high-fat diets (20% by weight) for one wk containing graded levels of erucic acid from 1 to 50%, to evaluate the effect of short-term feeding and interaction of male sex hormones on formation of heart lesions. Some rats within each group were returned to the low-fat diet for one wk after the test period. For comparison, one group of three-wk-old male rats was fed the high fat 50% erucic acid diet for 15 wk. Erucic acid depressed growth rate and food consumption and increased cardiac lipidosis and triglycerides proportional to the erucic acid content of the diet. There were no sex differences, and the effects disappeared once rats were returned to the low-fat diet for one week. There was a significance (P less than 0.05) in the incidence of myocardial necrosis among male rats fed increased levels of erucic acid for one week, but the response was not linear to the increase in dietary erucic acid. Furthermore, the response was much less than in males fed the 50% erucic acid diet continually for 15 weeks. These results suggest that the short-term model is not a suitable substitute for the long-term feeding trial to test the cardiopathogenicity of a vegetable oil. The significantly lower incidence in myocardial lesions in female and castrated male rats compared with male rats suggests involvement of sex hormones.(ABSTRACT TRUNCATED AT 250 WORDS)

Optimization of chick embryotoxicity bioassay for testing toxicity potential of fungal metabolites.

The optimization of a simple, sensitive procedure using a chick embryotoxicity screening test (CHEST) bioassay for detection of toxic compounds is presented. Dosing protocols of eggs, using several mycotoxins (aflatoxin B1, deoxynivalenol, T-2 toxin) and appropriate controls, were evaluated for embryonic sensitivity, overall practicality of the procedure, and consistency of results. It was found that both type of carrier solvent and volume injected could significantly affect overall embryonic mortality. The chick embryo was most sensitive to the effects of toxins and solvents after 1 or 2 days of incubation; a rapid decrease in response was observed as the age of the embryo at dosing increased. Following administration of the toxins just below the shell membrane by way of a small hole (less than 0.5 mm diameter) punched in the shell, a good dose-response (% mortality) could be obtained regardless of the site of injection (except directly into the yolk), although dosing via the air sac position resulted in a slightly better statistical outcome. Although some variations in calculated LD50 values were found among repeated assays, statistical analyses showed that the differences were not due to dosing protocol but to the variations in embryo sensitivities among batches of eggs. Thus, if standard reference toxins for comparison are run concurrently, the CHEST assay can prove to be a very satisfactory model, as well as having considerable flexibility to be adapted to the needs and resources of many laboratories.

Effects of feeding white Leghorn hens diets that contain deoxynivalenol (vomitoxin)-contaminated wheat.

A short-term (10 weeks, Experiment 1) and a long-term experiment (24 weeks, Experiment 2) were done to determine effects of incorporating either white winter wheat, naturally contaminated with 1 mg deoxynivalenol (DON)/kg, or spring wheats, containing up to 6.5 mg DON/kg, into the diets of White Leghorn hens. Based on chemical analysis, the diets in Experiment 1 contained less than .05 to .7 mg DON/kg, while those in Experiment 2 contained from .2 to 4.9 mg/kg. Incorporation of winter or spring wheat in the experimental diets had no effect (P greater than .05) on feed intake and efficiency, egg production and yield, the number of soft shell and cracked eggs observed in the laying house, body weight at the completion of the experimental period, fertility, hatchability of fertile eggs, and the proportion of malformed embryos and pips. In addition, presence of DON-contaminated wheat did not influence (P greater than .05) the organ weight to body weight ratio for a randomly selected sample of hens necropsied at the completion of each experiment. There was little evidence of lesions in the oral cavity, esophagus, proventriculus and gizzard, hemorrhaging in the viscera or skeletal muscles, or of changes in the appearance of spleen, heart, and kidney. However, the livers from DON hens were fatty in appearance. Furthermore, vomiting (emesis), diarrhea, or changes in behaviour were not apparent and mortality, normally very low, was not increased during either experiment. Inverse linear relationships were obtained in Experiment 1 between dietary DON concentrations and egg weight (P less than .05), shell weight and thickness (P less than .01), and percent shell (P less than .05). Although egg and shell variables measured in Experiment 2 were not significantly influenced (P greater than .05) by DON treatment, trends towards lower values with higher dietary DON levels were evident. Egg specific gravity, nondestructive deformation, and quasistatic compression fracture strength of the egg's shell were not influenced (P greater than .05) by dietary DON levels. The results from these experiments indicate that laying hens can tolerate diets containing up to 5 mg DON/kg from white winter or spring wheat for extended periods of time without serious adverse effects on health and productivity.

Cardiac lipid changes in rats fed oils enriched in saturates and their apparent relationship to focal heart lesions.

Weanling male Sprague-Dawley rats were fed diets containing 20% by weight corn, soybean or low erucic acid rapeseed oils or mixtures of the latter two with cocoa butter or triolein for 1, 2, 3 or 4 weeks. These diets previously had been fed to the same strain of rats for 16 weeks, and a reduction in the incidence of focal heart lesions had been observed with the addition of cocoa butter, but not triolein. The cardiac lipid classes and the fatty acid and alkenyl ethers of the cardiac phospholipids were analyzed to determine if changes could be attributed to the observed cardiopathological response, and at what time. Cardiac lipid classes changed during post-weaning development, but only triacylglycerol was diet-related. A number of fatty acid changes were observed in the cardiac phospholipids which reflected the relative concentration of saturates, monounsaturates, linoleic acid and linolenic acid in the diet, but only the changes in saturates and the C22 polyunsaturated fatty acids from the linolenic acid family appeared to be related to the incidence of focal heart lesions. Arachidonic acid and the total C22 polyunsaturated fatty acids remained fairly constant throughout the feeding trial. Cardiac diphosphatidylglycerol was least affected by dietary manipulation, while nervonic acid increased in cardiac sphingomyelin when small amounts of erucic acid were present in the diet. Fatty acid changes were essentially completed after one week on the experimental diets, whereas changes in the alkenyl ethers took two to three weeks.

Quantitating heart lipids: comparison of results obtained using the Iatroscan method with those from phosphorus and gas chromatographic techniques.

The precision and accuracy of the Iatroscan method was evaluated by comparing the results obtained with established phosphorus and gas chromatographic techniques. A complete lipid class analysis of rat heart lipids was chosen in order to evaluate the performance of the Iatroscan method for biological samples which contained both neutral lipids and phospholipids. A partial scan and repeat development with chloroform/methanol/water (68.5:29:2.5) was introduced to achieve consistently good separations of the phospholipids on the Chromarods in the Iatroscan method. The results showed that the precision of the Iatroscan method for some lipid classes was comparable to that of phosphorus or gas chromatographic techniques, while for other lipid classes it was lower. Compared to the data obtained using the phosphorus method, the Iatroscan data were generally similar, while the gas chromatographic method generally gave lower values. These findings, together with the advantages of time required for analysis, size of sample, and universality of detection, suggest that the Iatroscan is a valuable complementary method for complex lipid analyses.

Ingestion of vomitoxin (deoxynivalenol)-contaminated wheat by nonlactating dairy cows.

Our objective was to determine if there were serious deleterious effects of wheat naturally contaminated with vomitoxin (deoxynivalenol) on nonlactating dairy cows. Comparisons were between two Quebec spring wheat sources contaminated with Fusarium graminearum in a feeding trial involving 10 nonlactating Holstein dairy cattle offered good quality hay for ad libitum intake supplemented with wheat-oats concentrate (1 kg concentrate/100 kg body weight). Initially, for 3 wk all cows were fed hay plus wheat-oats containing 1.5 mg vomitoxin/kg. Each week thereafter two more cows, chosen at random, were switched to a second wheat-oats concentrate containing higher mycotoxin concentration (6.4 mg vomitoxin/kg). After the last two cows had been on the latter diet for 6 wk, all cows were returned to the previous wheat-oats concentrate containing the lower mycotoxin concentration (1.5 mg vomitoxin/kg). No unusual symptoms of illness that might be attributed to the mycotoxin-contaminated wheat were observed. In body weight gains, the data reflected a linear growth pattern with no apparent effect of changing from low to high vomitoxin content in the concentrate. Generally, consumption of the wheat-oats ration containing 6.4 mg vomitoxin/kg was lower than the 1.5 mg vomitoxin/kg concentrate. The progressive, linear decrease of hay consumption was not affected by source of contaminated wheat-oats diet.

The tolerance of White Leghorn and broiler chicks, and turkey poults to diets that contained deoxynivalenol (vomitoxin)-contaminated wheat.

Three experiments were done to determine the effects of giving White Leghorn chickens, broiler chickens, and turkey poults diets that contained white winter or spring wheat contaminated with 4-deoxynivalenol (DON). Diets that contained .35 and .7 mg DON/kg from winter wheat did not (P greater than .05) influence feed intake, body weight gain, feed/gain ratio, and organ weight/body weight ratios of Leghorn and broiler chickens, and poults when fed between 7 and 21 days in Experiment 1. Dietary DON levels up to 4.6 mg/kg increased the feed intakes (P less than .01) and daily body weight gains (P less than .05) of Leghorn chickens between 7 and 35 days of age in Experiment 2 but had little effect (P greater than .05) on these variables for broiler chickens between 7 and 52 days of age in Experiment 3. Within Experiments 2 and 3, feed/gain ratios and organ weight/body weight ratios were similar among the Leghorn and broiler chickens, respectively. The dressing percent and chilled carcass weights of eviscerated broiler chickens were not affected (P greater than .05) by DON in the diets of Experiment 3. Mortality was low (less than 3.3%) for the three experiments and necropsy examination indicated that the birds died from several causes unrelated to the dietary treatment. There was no evidence of emesis during the experiments or of overt changes in the appearance of the oral cavity, heart, liver, spleen, kidney, proventriculus, gizzard, and intestines of a random sample of birds killed by carbon dioxide asphyxiation and necropsied. Differences occurred between the analyzed and calculated DON contents of the experimental diets; possible reasons for these differences are discussed. The results of these experiments indicate that young chickens and turkey poults can tolerate diets that contain DON up to at least 5 mg/kg from white winter or spring wheat.

Feeding trials with vomitoxin (deoxynivalenol)-contaminated wheat: effects on swine, poultry, and dairy cattle.

Nutritional and toxicologic feeding trials with 3 species of farm animals demonstrated that decreased feed consumption and reduced weight gains in pigs are the main effects of ingestion of a diet with low vomitoxin (deoxynivalenol; DON) content, eg, 2 mg of DON/kg of feed, ie, 2 ppm. The feeding trials indicated that swine can ingest up to 2 mg of DON/kg of feed without serious adverse effects. Poultry can tolerate at least 5 mg of DON/kg feed. In fact, at concentrations up to 5 mg of DON/kg feed, some beneficial effects on poultry were observed. In dairy cattle, feed consumption decreased slightly when a wheat-oats diet containing 6 mg of DON/kg was fed at the rate of 1% of body weight/day, with hay offered ad libitum. In surveys of Canadian grains carried out during the past 3 years, the DON content (maximum of 8.5 mg/kg) in eastern Canadian wheats probably was not high enough to account for reports of feed refusal, vomiting, and reproductive problems in livestock operations. This conclusion is based partly on the fact that even at the highest concentrations of DON found in wheat, formulated diets comprise, at maximum, about 70% to 80% wheat. Consequently, the actual DON content of diets fed to farm animals would be much lower.