Hydrated sodium calcium aluminosilicate: a high affinity sorbent for aflatoxin.

Aluminas, silicas and aluminosilicates were evaluated for their ability to sorb radiolabeled aflatoxin B1 (AFB1) from aqueous solution (in vitro). Hydrated sodium calcium aluminosilicate (HSCAS) was selected for testing in vivo due to its high affinity for AFB1, because of its stable association with AFB1, and its GRAS (generally recognized as safe) status as an anticaking agent. The HSCAS, when added to the diet of Leghorn and broiler chicks at a level of .5%, significantly diminished the adverse effects of feeding 7.5 mg AFB1/kg of feed. Thus, this agent (and other aluminosilicate congeners) may prove effective in the preventive management of aflatoxicosis.

Determination of deoxynivalenol (vomitoxin) by high-performance liquid chromatography with electrochemical detection.

A rapid method for the analysis of deoxynivalenol (DON) was developed using high-performance liquid chromatography (HPLC) with reductive electrochemical detection (ED). Deoxynivalenol produced by Fusarium roseum growing on solid cornmeal and rice substrates and from naturally contaminated wheat was extracted and quantitated via ED. DON levels in wheat were verified by gas chromatography and structurally confirmed by mass spectrometry. DON was optimally resolved by HPLC employing a radially compressed octadecylsilane column and a mobile phase of deoxygenated methanol-40 mM borate buffer (35:65) at a flow-rate of 1.0 ml/min. Under these conditions DON exhibited an average retention time of 3.6 min. Reductive ED (-1.4 V) allowed a 12-fold increase in sensitivity and greater selectivity than classical UV absorption at 224 nm. A detection limit for DON of 25 pg/microliter was achieved under these conditions. The determination of DON in crude grain extracts was hindered by extractable interfering substances, whereas ED was more functional-group selective (i.e. reduction of the carbonyl moiety). ED permits a direct quantitation of DON from crude grain extracts and may facilitate the determination of this agent and associated metabolites in biological samples.

Effects of deoxynivalenol in a wheat ration fed to growing lambs.

A wheat diet containing deoxynivalenol (vomitoxin) at 15.6 mg/kg was fed to crossbred lambs for 28 days. Feed consumption, weight gain, and feed efficiency of deoxynivalenol-treated lambs did not differ (P less than 0.05) from those values of controls. Group differences were not noted for hematologic or serum biochemical variables, and gross or microscopic lesions were not observed in treated lambs.

Toxicity of ochratoxin A and vanadium to growing chicks.

The effects of ochratoxin A (OA) and vanadium (V), singly and in combination, were determined in male Leghorn chicks from 1 to 28 days of age. The chicks were fed a control diet containing the following additives: A) none; B) 2.5 mg OA/kg; C) 50 mg V/kg; D) 2.5 mg OA plus 50 mg V/kg. These data show that body weight gains were significantly reduced by OA and V singly, and a toxicity-enhancing synergism exists between OA and V, which caused a further reduction in performance. The OA-V combination caused a significant increase in the relative weights of the liver, kidney, gizzard, and proventriculus and a significant decrease in the relative weights of the bursa of Fabricius. This decrease in bursal weight was due to atrophy of lymphoid follicles as indicated by the increase in the histologic lesion score. Uric acid in serum was increased, and albumin, calcium, and phosphorus were decreased in the OA-V combination group. There were also small but significant changes in the hematological parameters measured. Distribution of OA to the liver and kidney was not altered, nor was the distribution of V to the liver, kidney, or muscle tissue. Histologic lesions in kidneys were mild but were more prevalent in chicks in the OA-V combination group.

Effects of ochratoxin A in the partially nephrectomized rat.

The effects of ochratoxin A (OA), a nephrotoxic mycotoxin, were investigated in partially nephrectomized (PN) rats (approximately 70% reduction in renal mass) following compensatory hypertrophy of the renal remnant. Renal function stabilized 27 d after surgery. PN rats compensated for the initial loss of renal function except for glomerular filtration rate (GFR, inulin clearance); this remained significantly impaired. Sham-operated (SO) rats cleared inulin and p-aminohippurate (PAH) at rates of 3.84 and 7.49 ml/min, respectively, while compensated PN rats cleared inulin at 2.51 and PAH at 8.84 ml/min. Daily administration of low levels of OA produced decreased urine osmolality and body weight with a modest increase in urinary protein of PN versus SO rats. OA-treated rats cleared inulin, creatinine, and PAH at rates significantly lower than nontreated controls: 0.89 and 1.96 ml/min for inulin, 0.35 and 0.56 ml/min for creatinine, and 2.29 and 6.23 ml/min for PAH. Histopathological findings indicated a considerable increase in renal tubular necrosis and subcellular damage (i.e., loss of cytoplasmic ground substance, vacuolization, degeneration of mitochondria, and reorganization of endoplasmic reticulum) in PN animals versus controls, concurrent with alteration in renal function. These results verify that the nephrotoxic action of OA is elicited mainly in renal proximal tubules and is enhanced in the PN rat.

Vanadium: chemistry and the kidney.

Vanadium (V), a metallic element of the first transition series, is widely distributed in the environment. Although an essential trace element in higher animals, chronic exposure to V is of concern because of its increased concentration near industrial operations, its occurrence in the ash of combustion products of petroleum and coal, and its subsequent biomagnification in the environment. V is found in trace amounts in both terrestrial and aquatic animals and in solution can form inorganic orthovanadate oxyanions that, if absorbed, are eliminated primarily by the kidneys. Because V is often found concentrated in renal tissue to the largest extent in the body, the kidneys may represent a major site of action. Moreover, V in the vanadate configuration increases the urinary excretion of solutes and water in the rat, and inhibits renal organic ion accumulation and renal Na+, K+-ATPase in vitro and in vivo. Furthermore, as a nutritionally required element, V may play a regulatory role in salt and water excretion by modification of the Na+ pump in the kidney.

Toxicity of ochratoxin A and tannic acid to growing chicks.

The effects of ochratoxin A (OA) and tannic acid (TA) on growing chicks were determined. One-day-old male broiler chicks were fed a diet containing the following additives for 26 days: A) none; B) 3.0 ppm OA; C) 1.5% TA; D) 3.0 ppm OA plus 1.5% TA. When compared to the controls, body weights and feed efficiencies were significantly depressed in the OA and TA groups. There was a further depression in body weights and a dramatic depression of feed efficiency in the OA-TA combination group. Pigmentation, as measured by visual shank scores, was reduced in chicks fed OA singly or in combination with TA but was not affected by feeding TA singly. There were no consistent treatment differences in the relative weights of the kidney, gizzard, proventriculus, liver, bursa, or pancreas, although there was a trend toward an elevated relative kidney weight in the groups receiving OA. Serum uric acid levels were significantly elevated in the OA and the OA-TA combination group indicating impaired renal excretory function attributable to OA. Total serum protein levels were significantly depressed in the groups receiving OA, and serum calcium levels were depressed in all treatment groups. Serum phosphorus levels were decreased in the OA and OA-TA groups but were only decreased significantly in the OA group. There were no consistent treatment differences in the hematology and other blood chemistry and mineral values.

High pressure liquid chromatographic determination of an O-methyl,methyl ester derivative of ochratoxin A.

The mycotoxin ochratoxin A (OA) was derivatized to an O-methyl,methyl ester (Me2) with diazomethane and then determined by high pressure liquid chromatography (HPLC). Both OA and OA-Me2 were chromatographed by reverse phase HPLC with a mobile phase of acetonitrile-water (60 + 40). An increase in retention time of 309 s was observed with OA-Me2 which was detectable at 254 nm at levels as low as 3 ng. Recovery of OA as OA-Me2 from chicken kidney homogenates and human plasma was quantitative following simple extraction and cleanup procedures, reaction with diazomethane, and HPLC analysis. The novel method described should prove useful for measuring and confirming OA in tissues and in further studies on the biological fate of this mycotoxin.

High pressure liquid chromatographic determination of zearalenone in chicken tissues.

A method is reported for the extraction and analysis of zearalenone in chicken fat, heart muscle, and kidney tissue by using high pressure liquid chromatography (HPLC). Zearalenone is extracted with acetonitrile, cleaned up with hexane, and extracted further with ethyl acetate. Zearalenone is determined by HPLC using a reverse phase radial compression separation system, an ultraviolet absorbance detector, and a mobile phase of acetonitrile-water (60 + 40) (v/v). Recoveries of zearalenone added at levels from 50 to 200 ng/g are in the range 82.6-95.1%.

Veterinary food hygiene in the year 2000: circumstances and needs.

A series of surveys was conducted among veterinarians engaged in food hygiene, public health, and/or preventive medicine practice. The objective was to identify the circumstances and needs in veterinary food hygiene for the year 2000. Circumstances were defined as the external factors that influence veterinary food hygiene (eg, perspectives, general technologic advances in society, social expectations). Needs were defined as the internal resources that are essential to the practice of veterinary food hygiene (eg, goals, evaluations, personnel, communications, technology). The results of the surveys could be useful in the design of innovations for veterinary food hygiene programs.

High pressure liquid chromatographic determination of aflatoxins by using radial compression separation.

A rapid method for the determination of aflatoxins was developed using high pressure liquid chromatography and a radial compression separation system. A standard solution of aflatoxins B1, B2, G1, G2, and M1 was analyzed solution of aflatoxins B1, B2, G1, G2, and M1 was analyzed at flow rates of 2.0 and 6.0 mL/min. Retention times, peak heights, and peak areas were reproducible over a 3-day period. Coefficients of variation for aflatoxin B1 at 2.0 and 6.0 mL/min were, respectively, 1.04 and 0.87% (retention time); 2.9 and 4.7% (peak height); and 8.2 and 4.7% (peak area). At 6.0 mL/min there was an approximate 25% loss in sensitivity but a greater than 50% reduction in retention time. Separation of all the aflatoxins was excellent using a dual flow rate of 2.0 mL/min with a change to 8.0 mL/min at 15 min post-injection. The applicability of the radial compression separation system for the rapid determination of aflatoxins in human tissues was also tested. Spiked samples of liver, serum, and urine-showed good resolution of all aflatoxin peaks at the higher flow rates.

Vanadium-induced inhibition of renal Na+, K+-adenosinetriphosphatase in the chicken after chronic dietary exposure.

Recent work has shown that V accumulates in the kidney and is a potent inhibitor of Na+, K+-adenosinetriphosphatase (ATPase) in vitro. Thus, as a nutritionally required element, V may regulate cation transport. The effect of chronic intake of the metal on Na+, K+-ATPase in vivo has not been reported. In this study laying strain chickens were fed calcium orthovanadate for 15 mo from d 1 of age at levels of 0, 25, 50, and 100 ppm in the diet. Whole tissue homogenates and 13,000 X g fractions were analyzed for ATPase activities. Concentrations of V producing 50% inhibition of Na+, K+-ATPase activity ranged from 1.0 X 10(-5) M in liver to 1.8 X 10(-6) M in kidney, which was the most sensitive tissue tested in vitro. Mg2+ -ATPase was more resistant to V than Na+, K+-ATPase. Studies in vivo suggested a V-dependent inhibition of renal Na+, K+-ATPase. Correlation of enzyme specific activity and levels of V in kidneys suggested V-ATPase mediated alteration in renal function.

Inhibition of pancreatic carboxypeptidase A: A possible mechanism of interaction between penicillic acid and ochratoxin A.

Penicillic acid and ochratoxin A are environmentally important toxic fungal metabolites (mycotoxins) that are synergistic in combination. The effects of penicillic acid on the pancreatic enzyme, carboxypeptidase A were investigated in vitro and in vivo. A broad range of inhibition in vitro of the enzyme by PA was demonstrated with a half-maximal inhibitory concentration equal to 1.1 x 10(-4) M PA. Inhibition of carboxypeptidase A was time and temperature dependent, and resulted in decreased conversion of parent ochratoxin A to the non-toxic metabolite, ochratoxin alpha. Studies in vivo demonstrated a penicillic acid-dependent inhibition of pancreatic carboxypeptidase A activity in the mouse and the chicken following multiple oral exposure. It is postulated that the mode of toxic interaction of the two mycotoxins may be due, in part, to impaired detoxification of ochratoxin A through penicillic acid depletion of carboxypeptidase A activity.