Improved spectrophotometric determination of cyclopiazonic acid in poultry feed and corn

An improved visible spectrophotometric method has been developed for cyclopiazonic acid in poultry feed and corn. The method is based on the reaction of cyclopiazonic acid with Ehrlich reagent and detection at 580 nm. Reaction conditions were optimized with respect to reaction and measurement times and acid and Ehrlich reagent concentrations. Calibration curves were linear from 1 to 20 micrograms cyclopiazonic acid in 3 mL Ehrlich reagent, with a lower detection limit of 0.08 mg/kg for 50 g samples of poultry feed and corn. Recoveries from 50 g samples of poultry feed spiked with cyclopiazonic ranging from 0.16 to 1.20 mg/kg averaged 93.8%. Moldy corn and poultry feed samples analyzed by this method contained between 1 and 4 mg/kg cyclopiazonic acid.     

Liquid chromatographic determination of nicarbazin in feed

A liquid chromatographic method was developed for the determination of nicarbazin (4,4'-dinitrocarbanilide.2-hydroxy-4,6-dimethylpyrimidine) in chicken feed. Ground feed was extracted with hot dimethylformamide, filtered, and then cleaned up on an alumina column. The nicarbazin was eluted from the column with ethanol and quantitated using a reverse phase C-18 column, with a methanol-water mobile phase and ultraviolet detection at 344 nm. Recoveries at a typical use level of 100 micrograms/g feed averaged 98% with a standard deviation of 3%. Samples fortified at levels as low as 0.1 micrograms/g were analyzed with 92% recovery. The detection limit is 1 ng, and the response is linear between 4 and 1000 ng. Feed additives in combination with nicarbazin do not interfere with recovery.     

Detection, accumulation, distribution, and depletion of furaltadone and nifursol residues in poultry muscle, liver, and gizzard

Nitrofurans were broadly used as an extremely effective veterinary antibiotic especially in pig and poultry production farms. Because of fears of the carcinogenic effects on humans, the nitrofurans were banned from use in livestock production in many countries, including the European Union. The present study examines the accumulation, distribution, and depletion of furaltadone and nifursol and of their tissue-bound metabolites [3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) and 3,5-dinitro-salicylic acid hydrazine (DNSAH), respectively, in poultry edible tissues (muscle, liver, and gizzards) following administration to chickens of therapeutic and subtherapeutic concentrations of both compounds. Nitrofurans determination was performed by high-performance liquid chromatography-diode array detection and liquid chromatography-tandem mass spectrometry, respectively, for feeds and for poultry tissues. Furaltadone and nifursol, in very low concentrations, were found in samples of muscle, liver, and chicken's gizzard collected from slaughtered animals after 5 weeks of treatment and no withdrawal time period. When a withdrawal time period of 3 weeks was respected, no detectable nitrofuran parent compounds was observed in all of the studied matrices. For AMOZ, concentrations of 270 μg/kg in meat, 80 μg/kg in liver, and 331 μg/kg in gizzard were determined after administration of a medicated feed with furaltadone (132 mg/kg), 3 weeks after withdrawal of treatment. For DNSAH, the concentration values obtained are much lower than those observed for AMOZ. For meat, liver, and gizzard, DNSAH concentrations of 2.5, 6.4, and 10.3 μg/kg, respectively, were determined, after administration of a medicated feed with nifursol (98 mg/kg), 3 weeks after withdrawal of treatment. The gizzard could be considered a selected matrix for nitrofuran residues evaluation in poultry, due to its capacity of retaining either nitrofuran parent compounds or metabolites in higher concentrations, regardless of the administered dose or of the respected withdrawal time period.     

浙江省重点地区猪粪中重金属含量及安全施用评估

【目的】 浙江的规模养殖集中度高,畜禽排泄物资源化利用率达96%以上,通过堆肥和制作有机肥还田是主要的消纳途径,因此粪便中的重金属含量影响土壤安全及作物的生长。饲料是猪粪重金属最主要的来源,因此,本研究检测了浙江省猪饲料和排泄物中的重金属,为作物施用的安全性评价提供基础数据。 【方法】 采集35家不同规模养殖场的仔猪、生长猪和育肥猪3个阶段饲喂的饲料和粪便样本共566个,采用干灰化法、高压消解法消解,用AAS、AFS、ICP-MS法分析了铁、铜、锰、锌、砷、铅、铬、镉和汞9种重金属含量。 【结果】 1) 粪便中8种重金属平均含量趋势为Zn > Cu > Mn > As > Cr > Pb > Cd > Hg,主要残留元素为Zn、Cu、Mn和As,按干物质计其平均值分别为2405、1288、528和28.2 mg/kg,变异系数 ( CV) 分别为83%、34%、6%和15%。猪粪中Cr、Pb、Cd元素含量较低,在0.29～3.52 mg/kg之间,Hg含量仅3.97 μg/kg。猪粪中重金属含量仔猪阶段趋高,育肥猪阶段趋低,各个阶段变异都较大,变异系数CV值介于21%～112%之间,显示各地的生猪饲养中微量元素添加还比较混乱。2) 饲料有机砷的使用存在明显区域差异,萧山地区占72.7%的饲料样本在各阶段猪饲料中均添加有机砷,衢州地区的占33.3%,两个地区各有1家养殖场未使用有机砷。猪粪中54.9%样本砷含量 > 15 mg/kg,16.9%样本砷含量 > 50 mg/kg,2.8%的粪便砷超过70 mg/kg。3) 按畜禽粪便安全使用准则进行判定,69.9%猪粪铜超标,16.9%猪粪砷超标。按有机肥料标准进行判定,猪粪中54.9%样本砷含量超标。按照饲料中金属元素在粪便中的浓缩率:Fe、Zn、Cu为7～10倍,锰约为3倍,当前饲养水平下,Fe、Zn、Cu元素存在过量添加。4) 以浙江全省旱地单位面积承载量估算,当前能消纳猪粪中Zn、Cu的排放,但当每公顷猪粪施用量超过2.27 t (干计) 时,局部地区重金属就有累积风险。 【结论】 应降低猪饲料中微量元素的添加量,禁止饲用有机砷制剂,才能有效降低猪粪施用所导致的重金属污染;按照目前监测猪粪肥重金属元素含量水平,有机肥的施用量用应控制在2.27 t/hm2 (干基) 以内,以防止对土壤造成污染。      

规模化养殖畜禽粪主要有害成分测定分析研究

随着我国规模化养殖业的快速发展,畜禽粪中重金属、兽药残留、盐分及有害菌等有害污染物的增加,构成影响农田土壤健康功能的限制因素,甚至带来生态环境风险。对我国7个省、市、自治区的典型规模化养殖畜禽粪的主要化学组成进行了测定。结果表明,规模化养殖畜禽粪中富含有机质、氮、磷、钾及钙、镁、铁、钠、锰、钴等矿质元素;砷、铜、锌、铬等重金属及盐分含量较高,55个猪粪、鸡粪样中,Cu、Zn、Cr、As含量变幅分别为10.7～1591mg/kg、71.3～8710.mg/kg、0～688.mg/kg、0.01～65.4.mg/kg。同时多有四环素类抗生素残留,32个猪粪样中,土霉素、四环素、金霉素平均含量分别为9.09.mg/kg、5.22.mg/kg、3.57.mg/kg;23个鸡粪样中,土霉素、四环素、金霉素平均含量分别为5.97.mg/kg2、.63.mg/kg1、.39.mg/kg。因此,规模化养殖畜禽粪对农田施用后存在潜在危害性,规模化养殖畜禽粪必须进行无害化处理,才能资源化利用。     

Rapid time-resolved fluoroimmunoassay for the screening of narasin and salinomycin residues in poultry and eggs

Anticoccidial drugs are extensively used in the poultry industry to control the infection of the single-cell protozoa of the genus Eimeria. The most commonly used coccidiostats in poultry are the polyether ionophores such as narasin and salinomycin. This paper presents a rapid and simple method for the screening of residues of these two coccidiostatic compounds in poultry and eggs. The method is based on time-resolved fluoroimmunoassay. Sample preparation of eggs consists only of one extraction and evaporation step, and a solid phase extraction step is needed only for the muscle sample preparation. Mean recoveries were 91.0% from muscle tissue and 81.1% from eggs for both narasin and salinomycin. The performance of the assay was evaluated only for narasin because salinomycin had a cross-reactivity of 100% in the assay, and the recoveries of the compounds were not significantly different (P >0.05). The limits of detection [mean + 3 x standard deviation (SD)] of narasin were 0.56 and 0.28 microg/kg, and the limits of quantification (mean + 9 x SD) were 1.80 and 0.57 microg/kg for muscle and eggs, respectively. The coefficients of variation (CV) of the interassay precision of the method, evaluated by five replicate analyses of muscle samples spiked with 2 microg/kg of narasin and egg samples spiked with 1 microg/kg of narasin, were 4.1 and 6.4%, respectively. The CVs of intra-assay precision tests, determined by 10 replicate analyses at the above-mentioned concentration levels, were 3.8 and 4.5%, respectively.     

High performance liquid chromatographic determination of clocapramine in feed and plasma

A high performance liquid chromatographic method is described for the determination of clocapramine in animal feed and plasma. Samples are made alkaline and then extracted with chloroform containing opipramol as internal standard. For plasma samples, the organic phase is evaporated to dryness under a stream of nitrogen, and the residue is dissolved in dichloromethane-methanol. Extracts are chromatographed on silica gel with dichloromethane-methanol-ammonia (100 + 10 + 0.25) as eluant, and quantitated using an internal standard. Within-day precision for plasma extracts (n = 15) was 3.39, 5.7, and 4.13% at 5, 10, and 15 mg clocapramine/L plasma, respectively, and day-to-day precision was 4.6, 6.8, and 4.4% at the same levels. The detection limit was 0,5 mg/L. Recovery from feed over the concentration range 2-6 g/kg was greater than 96%.     

Thin-layer chromatography/bioautography method for detection of monensin in poultry tissues

Monensin is an effective anticoccidial agent widely used in the poultry industry. Because of concerns over its toxicity, a sensitive, reliable, fast, and simple method for residue detection in poultry tissues is desirable from both a diagnostic and a regulatory view. Many methods of detection are excluded due to this ionophore's complex chemical and biological nature. A common method used for its detection is thin-layer chromatography/bioautography (TLC/B), although this is usually only semiquantitative. A new TLC/B method for monensin detection in poultry tissues was developed and is reported here. Recovery from cardiac muscle, skeletal muscle, and liver and kidney tissues is in the range of 93-97%. A detection limit of 250 micrograms/kg with 99% reliability was achieved with this method. Lower limits (to 10 micrograms/kg) were detectable, but with lower reliability (60%). Quantitative analysis is not possible on samples from fatty tissue.     

Deposition and depletion of five anticoccidials in eggs

Anticoccidials are compounds that are widely used as feed additives to prevent and treat coccidiosis. They are licensed for use in a prescribed concentration and during a certain time interval for broilers and pullets but not for laying hens. It was shown in the past that carry-over at the feeding mill is found to be the main reason for the presence of residues in eggs. An animal experiment was set up to investigate the effect of carry-over at the feeding mill on the presence of residues of anticoccidials in eggs. For the compounds diclazuril, robenidine, halofuginone and nicarbazin in combination with narasin, two concentration levels were tested: the maximum allowed concentration for broilers (100%) and a concentration corresponding to 5% carry-over during feed preparation. Also dimetridazole was included in the experiment but only at one concentration level. Eggs were sampled during treatment (14 days) and for a period of 30 days after withdrawal of the anticoccidial-containing feed. Residues were determined, and deposition and depletion curves were generated. Analyses were performed by ELISA and LC-MS/MS. For all compounds, substantial residues could be found in the 5% groups, which points out the risk of carry-over at the feeding mill. The distribution of the residues between egg yolk and white was determined by analyzing both fractions.     

不同水平阿散酸粪肥对水稻生长发育及砷积累的影响

【目的】 饲料添加剂阿散酸是一种有机砷制剂,在动物肠道内吸收较少,绝大部分以原形随粪便排出体外。阿散酸中的砷可以随施用粪肥进入农田,对作物生长发育产生不利影响,并影响人类健康。本文研究了阿散酸进入土壤后,其所含砷在作物体内的迁移和积累,为粪肥合理施用提供依据。 【方法】 以水稻进行了盆栽模拟试验,供试土壤为褐土。将含有一定量阿散酸的动物粪便与供试土混匀,制成含阿散酸分别为30、75和150 mg/kg的试验用土样,按照每公顷22000 kg施用量,以无阿散酸污染的动物粪便为对照。在水稻分蘖期和开花期取样,分为根、茎、叶,成熟期样品分为根、茎、叶、稻壳和糙米,测定砷的含量和积累量。 【结果】 1) 与对照相比,30～150 mg/kg阿散酸各处理水稻根系长度、单株根数和株高无显著差异,150 mg/kg处理无效分蘖显著增加 (P < 0.05),75 mg/kg和150 mg/kg处理稻米产量显著降低 ( P < 0.05);2) 与对照相比,阿散酸各处理水稻分蘖期、开花期和成熟期根、茎和叶中砷含量显著增加 ( P < 0.05);与30 mg/kg阿散酸处理相比,150 mg/kg处理分蘖期、开花期和成熟期水稻根中砷含量显著增加了40.51%、46.25% 和53.1% ( P < 0.05),开花期和成熟期茎和叶中砷含量显著增加了56.1%、30.9% 和86.7% ( P < 0.05)、61.75% ( P < 0.05);阿散酸各处理水稻根中砷含量均在开花期达到最高,成熟期下降,阿散酸含量 ≤ 75 mg/kg时,水稻茎和叶中砷含量在开花期达到最高,成熟期下降,阿散酸添加量为150 mg/kg时,水稻茎和叶中砷含量为成熟期 > 开花期 > 分蘖期。3) 与对照相比,阿散酸处理籽粒砷含量显著增加 ( P < 0.05);与30 mg/kg阿散酸相比,75 mg/kg处理米糠和糙米中砷含量显著增加了138.5% 和126.1% ( P < 0.05),150 mg/kg处理颖壳、米糠、糙米和精米砷含量显著增加了24.6%、165.7%、158.7%和125.0% ( P < 0.05)。 【结论】 在褐土中施用含阿散酸非堆沤粪肥时,当阿散酸含量 ≥ 75 mg/kg时即对水稻产生毒害作用,并增加可食部砷污染风险,建议土壤阿散酸污染不得高于该值。      

Determination of natamycin in cheese and cheese rind: interlaboratory collaborative study

A collaborative test on the determination of natamycin in cheese and cheese rind was conducted. Participants were from 37 laboratories in 13 countries. Eight samples, consisting of 4 duplicates, were investigated by a spectrometric method and a liquid chromatographic (LC) method. The spectrometric method gave good results (coefficient of variation [CV] = 12%) and the LC method with ultraviolet detection gave reasonable results (CV = 25%) for levels down to 15 mg/kg (0.9 mg/dm2). For very low levels, a preconcentration step is necessary, but even then quantitation is poor (CV = 35-37%) for both methods at 1.7 mg/kg, although the presence of natamycin can be detected qualitatively. For a level of 0.3 mg/kg, quantitation is poor (CV = 39%) for the LC method and impossible (CV = 60%) for the spectrometric method.     

Deoxynivalenol and zearalenone residues in eggs of laying hens fed with a naturally contaminated diet: effects on egg production and estimation of transmission rates from feed to eggs

The potential for the Fusarium mycotoxins 4-deoxynivalenol (DON) and zearalenone (ZON) to enter the human food chain through contaminated eggs was assessed using a controlled feed study. Four groups of laying hens (eight in each group) were fed a diet that included differing amounts of naturally contaminated wheat containing DON ( approximately 20 mg kg(-1)) and ZON (0.5 mg kg(-1)). Eggs were collected and pooled from each group on a daily basis. Pooled samples were analyzed by liquid chromatography with mass spectrometry detection (LC-MS/MS). The method allowed DON, other type B trichothecenes, ZON, and its metabolites to be determined in a single multi-residue analysis. The selectivity of the MS/MS procedure allowed cleanup to be minimized (for DON, cleanup by immunoaffinity column was used) or eliminated (for ZON). The limits of detection of 0.01 microg kg(-1) for DON and 0.1 microg kg(-1) for ZON in eggs were lower than previously published methods. None of the samples analyzed had detectable levels of ZON or its metabolites. Although maximum levels of DON contamination (10 mg kg(-1) feed) were relatively high, no adverse effects were observed on egg production. On the basis of the determined DON levels in the hen's diet and the determined levels of DON in the corresponding eggs, transmission rates of 15 000:1, 18 000:1, and 29 000:1 for treatment levels 5, 7.5, and 10 mg DON kg(-1) feed, respectively, were found. These results show that, although eggs could be a human exposure route for DON, the levels are insignificant compared to the other sources, although the presence of metabolites of DON was not studied.     

Comparison of two immunochemical methods with thin-layer chromatographic methods for determination of aflatoxins

Three different methods were compared for the determination of total flatoxins in corn and peanuts naturally contaminated with aflatoxins and in corn, peanuts, cottonseed, peanut butter, and poultry feed spiked with aflatoxins B1, B2, and G1. The 3 methods were an enzyme-linked immunosorbent assay (ELISA) screening test; a monoclonal antibody-affinity column-solid-phase separation method; and the AOAC official thin-layer chromatography (TLC) methods for all except poultry feed, for which Shannon's TLC method for mixed feed was used. The ELISA test is designed to provide only positive results for total aflatoxins at greater than or equal to 20 ng/g or negative results at less than 20 ng/g. The affinity column separation is coupled with either bromination solution fluorometry to estimate total aflatoxins or liquid chromatography (LC) to quantitate individual aflatoxins. Fluorodensitometry was used to determine aflatoxins in commodities analyzed by the TLC methods. The LC and TLC results were in good agreement for all the analyses. The results for the affinity column using bromination solution fluorometry were similar except those for cottonseed, which were about 60% higher. The ELISA screening method correctly identified naturally contaminated corn and peanut positive samples. No false positives were found for controls. The correct response for spiked corn, raw peanuts, peanut butter, and cottonseed at greater than or equal to 20 ng aflatoxins/g was about 90%. The correct response for spiked poultry feed at greater than or equal to 20 ng aflatoxins/g was about 50%.     

Detection of residues of the coccidiostat diclazuril in poultry tissues by liquid chromatography-tandem mass spectrometry after withdrawal of medicated feed

A liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the quantitative determination of diclazuril in poultry tissues and feed is presented. A simple clean up with an organic solvent was carried out. A reversed-phase C(18) column was used for the high-performance liquid chromatography (HPLC) to separate the analyte with a gradient of acetonitrile and water as mobile phase. The precursor ions produced by electrospray negative ionization were selected for collisional dissociation. Validation of the methods was performed based on Commission Decision 2002/657/EC (Off. J. Eur. Communities 2002, L221, 8-36). For the detection of diclazuril in poultry meat, the decision limit was found to be 0.5 microg/kg. An animal experiment was set up in which 70 chickens were held for 6 weeks. From day 22 until day 32, they were fed feed containing 730 microg/kg diclazuril. From day 33 until day 42, every day six chickens were slaughtered, and breast, thigh, and liver were analyzed. Average steady-state concentrations of 94, 135, and 722 microg/kg in breast, thigh, and liver were obtained, respectively. Nine days after withdrawal of the medicated feed, diclazuril was still present in the different sample types.     

陕南茶园茶叶品质分析及重金属含量现状评估

为了解陕南茶园茶叶品质及茶园重金属含量现状,以茶叶主产区为研究对象,采集了8个县区330个茶园土壤样品,测定分析了pH值和铅(Pb)、锌(Zn)、铜(Cu)、铬(Cr)、汞(Hg)、砷(As)、镉(Cd)等7种重金属含量;同时测定分析了33个茶园茶叶品质及茶叶和浸出液中重金属含量。结果表明,茶叶中茶多酚平均含量为25.58%,儿茶素平均总量为20.74%,茶氨酸含量均值为2.45%,游离氨基酸总量均值为4.86%。茶叶中各重金属含量差异较大,Pb为0.04～2.65 mg/kg、Zn为34.82～55.38 mg/kg、Cu为3.18～20.16 mg/kg、Cr为0.09～4.89 mg/kg、Hg为0.02～0.30 mg/kg、As为0.66～1.16mg/kg和Cd为0.02～0.19mg/kg;32茶园茶叶符合《无公害食品茶叶》(NY5244—2004)和NY659—2003标准,33个茶园茶叶Cu含量符合有机茶标准。浸出液中仅检测出Zn、Cu和Cr,平均浸出率分别为:32.65%、24.96%和10.50%。土壤中重金属Pb、Zn、Cu、Cr、Hg、As和Cd的平均含量分别为10.03、87.61、16.42、12.38、0.20、6.89和0.11 mg/kg,综合潜在生态风险指数平均值为95.4,潜在生态风险较低。     

Residue levels of chlorpropham in individual tubers and composite samples of postharvest-treated potatoes

Chlorpropham, a herbicide and sprout suppressant, is used on stored potatoes to prolong the storage period without deterioration of produce quality. Data for residue concentrations on an individual tuber basis are required by WHO for the estimation of the variability factor. In this study, the levels of chlorpropham in individual tubers and in composite samples were determined. The distribution of chlorpropham between the peel and the tuber flesh was examined, and the fate during the cooking process (washing, boiling, frying) was studied. The concentrations in individual tubers ranged from 1.8 to 7.6 mg/kg 10 days postapplication (mean 3.8 mg/kg, RSD 39%), from 0.7 to 4.0 mg/kg 28 days postapplication (mean 2.9 mg/kg, RSD 28%), and from 0.8 to 3.8 mg/kg 65 days postapplication (mean 2.2 mg/kg, RSD 48%). The calculated residues in composite samples 10 days postapplication ranged from 4.3 to 6.1 mg/kg (mean 4.9 mg/kg, RSD 20%). Those in samples taken 28 days postapplication ranged from 3.1 to 4.2 mg/kg (mean 3.8 mg/kg, RSD 15%). The concentrations determined in composite samples of whole tubers 65 days postapplication ranged between 2.6 and 3.2 (mean 2.9 mg/kg, RSD 11%). Peeling removed 91-98% of the total residue; washing reduced residues by 33-47%. Detectable residues were found in boiled potatoes and the boiling water, and in french fries and the frying oil. Monitoring data on commercial prefried frozen french fries are reported.     

A new high-performance liquid chromatography-tandem mass spectrometry method based on dispersive solid phase extraction for the determination of the mycotoxin fusarin C in corn ears and processed corn samples

Fusarin C is a mycotoxin that is produced by a variety of Fusarium species and is therefore a possible contaminant in food and feed. For this reason, a reliable high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the determination of fusarin C in food and feed samples was developed based on dispersive solid phase extraction (DSPE). This method has a limit of detection (LOD) of 2 μg/kg, a limit of quantitation (LOQ) of 7 μg/kg, and a recovery rate of 80%. Fifty different corn samples were analyzed, and fusarin C was detected in 40 of them. The fusarin C level varied in kernels of corn ears from not detectable up to 83 mg/kg and in food samples from not detectable up to 28 μg/kg. The co-occurrence of further structural analogues of fusarin C was confirmed by high-performance liquid chromatography Fourier transformation mass spectrometry (HPLC-FTMS). In addition, the stability of fusarin C under storage conditions was evaluated.     

Alterations in yield,physicochemical components and mineral composition of onion following organic manure and inorganic nitrogen application

Onion is the one of the most important vegetable crops grown extensively throughout the world; hence, understanding the response of the crop to the form and amount of N provided is of immense importance. In a field experiment, poultry and cattle manure (at 10?t/ha) were compared with four inorganic nitrogen (N) application levels (0, 60, 90 and 120?kg?N/ha), and six combinations of manure and N applications were investigated. Plant growth and total yield (kg/ha) significantly increased, when N application increased from 0 to 120?kg/ha, with or without added poultry manure. Plant height, leaf number and neck thickness were significantly reduced at 0?kg/ha?N plus cattle manure. The bulb ascorbic acid concentration significantly increased with the use of poultry manure (15.94?mg/100?g) and cattle manure (15.94?mg/100?g), compared with 120?kg/ha inorganic N (11.42?mg/100?g). No significant effects were observed on total phenolics and total soluble solids of onion bulbs following the different fertiliser treatments. Cattle manure increased P and K concentrations, whilst poultry manure significantly increased Zn and Fe concentrations in onion bulbs. Onion bulb Ca and Mg concentrations were lower when cattle manure (0.19% and 0.12%) was used than when other organic or inorganic fertilisers were used. Therefore, organic manure increases the concentrations of certain minerals in onions, compared with inorganic N application, without negatively affecting bulb size.     

Fumonisin contamination and fusarium incidence in corn from Santa Catarina,Brazil

In Brazil, the southern region has the highest incidence of esophageal cancer and also the highest production and consumption of corn (Zea mays) products. Corn samples intended for human consumption from the western, northern, and southern regions of the state of Santa Catarina, southern Brazil, had mean total fumonisin B (B(1), B(2), and B(3)) levels of 3.2, 3.4, and 1.7 mg/kg, respectively. Fusarium verticillioides, the predominant fungus in the corn samples, had mean incidences (percent of kernels infected) of 14, 11, and 18% for the three regions, respectively. Additional corn samples intended for animal feed from the southern region had a mean total fumonisin level of 1.5 mg/kg and a mean F. verticillioides incidence of 10%. The fumonisin levels in corn from the state of Santa Catarina, Brazil, were similar to the high levels determined in other high esophageal cancer incidence regions of the world.     

Production, characterization, and cross-reactivity studies of monoclonal antibodies against the coccidiostat nicarbazin

A cELISA was developed for the coccidiostat nicarbazin. On the basis of previous computer-assisted molecular modeling studies, p-nitrosuccinanilic acid (PNA-S) was selected as a hapten to produce antibodies to 4,4'-dinitrocarbanilide (DNC), the active component of the coccidiostat nicarbazin. Synthesis is described for the hapten [p-nitro-cis-1,2-cyclohexanedicarboxanilic acid (PNA-C)] used in a BSA conjugate as a plate coating antigen. Monoclonal antibodies (Mabs) were isolated that compete with nicarbazin, having IgM(kappa) isotype. Because of the lack of water solubility of nicarbazin, N,N-dimethylformamide (DMF) (3%, v/v) and acetonitrile (ACN) (10%, v/v) were added to the assay buffer to achieve solubility of nicarbazin and related compounds. The Nic 6 Mabs had an IC(35) value for nicarbazin of 0.92 nmol/mL, with a limit of detection of 0.33 nmol/mL. Nic 6 exhibited high cross-reactivity for PNA-S and PNA-C, and 3-nitrophenol, 4-nitrophenol, and 1-(4-chlorophenyl)-3-(4-nitrophenyl) urea. However, Nic 6 had little or no cross-reactivity with 15 other related compounds.