Survey of dioxin-like compounds in dairy feeds in the United States

The United States Environmental Protection Agency (USEPA) has completed a survey of dioxin-like compounds (including 17 dioxin and furan (CDD/F) congeners and 12 coplanar polychlorinated biphenyl (PCBs) congeners) in dairy feeds from 10 dairy research facilities around the United States, sampling the overall mixtures and the major and minor feed components. Low levels of dioxin were found in all feed mixtures with an average concentration of 0.05 pg/g (ppt) toxic equivalent (TEQ) dry weight. This is lower than previously found in dairy feeds by about a factor of 4. While it is possible that generally lower levels of dioxins in the environment in recent years may explain this result, examinations of the data suggest that the oven drying used to prepare the wet feed samples could have resulted in a loss of dioxins from the feed materials. The percentage of the total TEQ due to CDD/Fs was about four times that of PCBs. Leafy vegetations in the feed (the silages and the hays) had concentrations about twice as high as nonleafy, protected vegetation of the feeds (the ground or meal corn, cottonseed, and others). Minor components did not significantly influence the final feed mixture concentration of dioxin TEQ. However, in one of the feed mixtures, a minor component with a high concentration of 38.5 ppt TEQ effectively doubled the concentration of the overall feed mixture.     

Detection of ruminant meat and bone meals in animal feed by real-time polymerase chain reaction: result of an interlaboratory study

The commercialization of animal feeds infected by prions proved to be the main cause of transmission of bovine spongiform encephalopathy (BSE). Therefore, feed bans were enforced, initially for ruminant feeds, and later for all feeds for farmed animals. The development and validation of analytical methods for the species-specific detection of animal proteins in animal feed has been indicated in the TSE (Transmissible Spongiform Encephalopathies) Roadmap (European Commission. The TSE (Transmissible Spongiform Encephalopathy) roadmap. URL: http://europa.eu.int/comm/food/food/biosafety/bse/roadmap_en.pdf, 2005) as the main condition for lifting the extended feed ban. Methods based on polymerase chain reaction (PCR) seem to be a promising solution for this aim. The main objective of this study was to determine the applicability of four different real-time PCR methods, developed by three National expert laboratories from the European Union (EU), for the detection and identification of cattle or ruminant species in typical compound feeds, fortified with meat and bone meals (MBM) from different animal species at different concentration levels. The MBM samples utilized in this study have been treated using the sterilization condition mandatory within the European Union (steam pressure sterilization at 133 degrees C, 3 bar, and 20 min), which is an additional challenge to the PCR methods evaluated in this study. The results indicate that the three labs applying their PCR methods were able to detect 0.1% of cattle MBM, either alone or in mixtures with different materials such as fishmeal, which demonstrates the improvement made by this technique, especially when compared with results from former interlaboratory studies.     

Determinations of residual furazolidone and its metabolite, 3-amino-2-oxazolidinone (AOZ), in fish feeds by HPLC-UV and LC-MS/MS, respectively

The antibacterial drug furazolidone belonging to the group of nitrofuran antibacterial agents has been widely used as an antibacterial and antiprotozoal feed additive for poultry, cattle, and farmed fish in China. During application a large proportion of the administered drug may reach the environment directly or via feces. Although the use of furazolidone is prohibited in numerous countries, there are indications of its illegal use. It is known that furazolidone can be rapidly metabolized to 3-amino-2-oxazolidinone (AOZ) in the body of the target organism. In this study, a total of 21 fish feed samples, including 17 commercial fish feeds from local markets in China (representing 15 different formulations) and 4 fish feeds obtained from Germany and Turkey, respectively, are analyzed to determine whether the drug is still illegally used or commercially available feeds are contaminated by this drug. High-performance liquid chromatography (HPLC) and liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) methods have been implemented to determine furazolidone and its metabolite AOZ in fish feeds containing animal protein, respectively. An efficient and convenient cleanup method for the determination of furazolidone in fish feeds is developed, and a simple cleanup method for the determination of AOZ is used. Method recoveries for samples used were determined as 87.7-98.3% for furazolidone at two spike levels of 2.0 and 5.0 ng g-1 and as 95.6-102.8% for AOZ at spike levels of 0.4 and 0.8 ng g-1. Limits of detections were 0.4 ng g-1 for furazolidone and 0.05 ng g-1 for AOZ. The established methods are therefore suitable for the determination of furazolidone and its metabolite AOZ in fish feeds at trace contamination levels. Using the established methods, all fish feed samples have been proved to be furazolidone negative; however, AOZ is tested in 16 of 17 fish feeds obtained from local markets in the Hubei province of China, with a positive rate as high as 94.1%.     

Extraction and thin layer chromatography of aflatoxin B1 in mixed feeds

A method was developed for the determination of aflatoxin B1 in commercially prepared feeds. The method incorporates methylene chloride and citric acid solution extraction, cleanup on a small silica gel column, and thin layer chromatography for quantitation. Commercial turkey starter, catfish chow, medicated pig starter, broiler finisher, rabbit chow, horse feed, rat chow, and dog chow were investigated. The feeds were spiked with naturally contaminated corn at 4 different levels of aflatoxin B1 (16-130 microgram/kg). Three assays were run on each of the 32 combinations of feed and levels of aflatoxin. Mean recoveries were 85.9-92.8% at levels of 16.5, 32.9, 65.8, and 131.6 micrograms/kg. The relative standard deviation per assay was 18.6%. This method is more rapid and less involved than most previously published methods for mixed feeds.     

High-speed liquid chromatographic determination of monensin, narasin, and salinomycin in feeds, using post-column derivatization

A high-speed liquid chromatographic (LC) method using post-column derivatization is described for the determination of monensin, narasin, and salinomycin in a variety of animal feeds. The ionophores are extracted with hexane-ethyl acetate (90 + 10). A portion of the sample is evaporated, diluted to a known volume, and analyzed using a 6 cm 3 microns C18 column and an absorbance detector after post-column reaction with vanillin. The method has been applied to poultry and swine feeds with levels of 3-100 ppm added antibiotic. A comparison was also carried out with medicated poultry feed and beef feed lot supplement samples previously analyzed by 2 separate bioassay methods for monensin and salinomycin, respectively. Recoveries for the LC method ranged from 92.1 to 103% with an average recovery of 98.1% and a coefficient of variation of 3.65%.     

Aflatoxins in domestic and imported foods and feeds

Aflatoxins, metabolic products of the molds Aspergillus flavus and A. parasiticus, may occur in foods and feeds. These toxins cannot be entirely avoided or eliminated from foods or feeds by current agronomic and manufacturing processes and are considered unavoidable contaminants. To limit aflatoxin exposure, the U.S. Food and Drug Administration (FDA) has set action levels for these toxins in foods and feeds involved in interstate commerce. FDA continually monitors food and feed industries through compliance programs. This report summarizes data generated from compliance programs on aflatoxins for the fiscal year 1986. Commodities sampled included peanuts and peanut products, corn and corn products, tree nuts, cottonseed, milk, spices, manufactured products, and miscellaneous foods and feeds. Correlations were highest between aflatoxin contamination and geographical areas for corn/corn products and cottonseed/cottonseed meal. Higher incidences of aflatoxin contamination in corn and corn products designated for human consumption were observed in samples collected in the southeastern states (32 and 28%, respectively). A higher incidence of contamination was observed in corn designated for animal feed from Arkansas-Texas (74%) than from the southeastern states (47%). Only 3% of feed corn from corn belt states contained detectable aflatoxins. All aflatoxin-contaminated cottonseed was collected in the Arizona-California area; 80% of cottonseed meal analyzed from this area also contained detectable levels of aflatoxins.(ABSTRACT TRUNCATED AT 250 WORDS)     

Improved turbidimetric assay of tylosin in premix and animal feeds not containing urea

A turbidimetric method is described for the determination of tylosin in premix and animal feeds not containing urea. This method includes several modifications of existing tylosin turbidimetric and AOAC plate assays to remove interferences from the feed, to concentrate low levels of tylosin, and to reduce the variability of the assay results. An acidic alumina column cleanup step has been incorporated into the method to remove interferences from feed ingredients. A disposable C18 column was used to concentrate the tylosin from low-level feeds, and the use of a larger analytical sample size has decreased the variability of the assay results. Average recoveries of tylosin added to chicken and swine rations were 98 and 101%, respectively.     

Turbidimetric assay of tylosin in animal feeds containing urea

A turbidimetric method is described for determination of tylosin in animal feeds containing urea. This method includes several modified or new steps to existing turbidimetric and AOAC plate assays that improve the extraction of tylosin, remove interferences from feeds, free tylosin activity, concentrate tylosin from low-level feeds, and reduce variability of assay results. A larger analytical sample size has been incorporated into the assay to decrease variability of assay results. A methanol-phosphate buffer extraction solution has replaced the hot buffer and methanol extraction solution. A hydrolysis step, which is not contained in the AOAC plate assay, was developed to free tylosin from the tylosin urea adduct that forms over time in feeds containing urea. A disposable C18 column was used to concentrate tylosin from feeds at levels less than 15 ppm. By increasing the analytical sample size from 25 to 100 g, the coefficient of variation for 12 weighings of cattle feed was reduced from 28.4 to 9.3%. Average recoveries from cattle rations containing tylosin at levels of 8, 10, and 100 ppm were 94, 94, and 91%, respectively.     

Thin layer chromatographic determination of citrinin.

A thin layer chromatographic (TLC) method is described for the determination of citrinin in feeds. Citrinin is extracted from feed with methanol and water, the mixture is made alkaline with 10% sodium carbonate, and the aqueous solution is filtered and extracted with chloroform to remove most of the interfering materials. The aqueous layer is acidified with 2N HCl and extracted with chloroform. The chloroform extract is concentrated and spotted on a thin layer chromatographic (TLC) plate which is developed in chloroform-acetone-ethanol-water (60 + 40 + 10 + 1). The citrinin is viewed under ultraviolet light after TLC. Either visual or fluorodensitometric quantitation is used. Recoveries of citrinin from various feed samples spiked at levels of 2.0--5 micrograms/g were 75--92%. The proposed method can detect 0.5 micrograms/g feed, including corn, silage, ready mixed feeds, and feed pellets.     

Rapid automated turbidimetric assay for chlortetracycline hydrochloride, using Leuconostoc mesenteroides as the test organism.

A rapid turbidimetric assay has been developed for chlortetracycline hydrochloride (CTC-HCl) in finished animal feeds and feed supplements, using Leuconostoc mesenteroides as the test organism. Two modifications are presented: The incubation period of modification 1 is 2.5 hr and the sensitivity of the assay is 0.03 microgram CTC-HCl/assay tube. Modification 2 has a sensitivty of 0.01 microgram CTC-HCl/assay tube and requires an incubation period of 3.5 hr. For 21 feed formulations, the turbidimetric method recovered 95.7% of label claim. Recoveries of CTC-HCl standards from the same feeds ranged from 93.4 to 134% with a mean of 103%. The relative standard deviation among day-to-day duplicates is 3.50% for the faster modification and 1.63% for the more sensitive modification.     

重金属在猪饲料-粪便-土壤-蔬菜中的分布特征研究

本文调查分析了江西省余江县39个大型养猪场的饲料、猪粪、以及长期施用这些猪粪的菜地土壤及蔬菜的Cu、Zn、Pb、Cd含量,并对饲料、猪粪、土壤和蔬菜重金属含量进行了相关分析。结果表明,大猪和小猪饲料Cu含量超标率分别达81.6%和30.8%,Zn含量超标率分别达89.5%和94.9%,而Pb、Cd未超标。猪粪Cu、Zn含量亦严重超标,且饲料和猪粪中Cu、Zn、Pb、Cd含量呈显著正相关关系。土壤总Zn和总Cd含量分别有7.8%和5.2%的样品超过三级标准,污染较为严重。所有蔬菜样品Cu、Zn、Pb含量均未超过我国食品卫生质量标准,空心菜和芋头Cd含量超标。土壤总Cu、Zn、Cd含量与提取态呈显著正相关。     

Collaborative study of a spectrofluorometric method for lasalocid sodium in feeds.

A collaborative study has been completed on a spectrofluorometric method for determining lasalocid sodium in finished poultry feeds. After a brief treatment of feed samples with pH 4.7 buffer, the drug is extracted with ethyl acetate. The ethyl acetate extract is further cleaned up by treatment with HCI and NaOH and quantitatively measured by spectrofluorometry. Nonspecific fluorescence is corrected for via the lasalocid-boric acid complex. The method is rapid, sensitive, and free of interference. A number of other feed additives, including monensin and ethoxyquin, do not interfere. Nine collaborators participated in the study and agreement between laboratories was satisfactory. The average recoveries of lasalocid sodium from the mash feeds added at levels of 0.0064, 0.0080, and 0.0096% were 100, 98, and 99%, respectively, and the corresponding coefficients of variation were 12.8, 8.2, and 8.0% (all results included). The method has been adopted as official first action.     

Liquid chromatographic determination of oxfendazole in swine feeds

A relatively simple analytical method is presented for determination of oxfendazole (2-(methoxycarbonylamino)-5-phenylsulfinyl-benzimidazole) at levels as low as 0.012% in swine feeds, using cation exchange liquid chromatography (LC). The sample was extracted with a solvent mixture of methanol-glacial acetic acid (90 + 10) at 45 degrees C, using a gyrorotory shaker. Plant pigments and other feed excipients were removed using zinc acetate treatment and pH-controlled extraction. Oxfendazole was further separated from the remaining interferences and quantitatively determined by LC on a Partisil SCX column with acetonitrile-0.01M phosphate buffer as mobile phase. The method is stability-specific, linear, precise, and accurate at 80-120% labeled strength (relative standard deviation 0.9-1.7 with mean recovery of 98-99%). Supporting data at a level of 0.0135% oxfendazole in swine feed indicated that this method is capable of complete recovery of oxfendazole from medicated swine feeds.     

Determination of aflatoxins, ochratoxin a, and zearalenone in mixed feeds, with detection by thin layer chromatography or high performance liquid chromatography

A sensitive, reliable, and economical method for the determination of 6 mycotoxins in mixed feeds is described. The feed is extracted with chloroform-water and the extract is cleaned up by using a disposable Sep-Pak silica cartridge. The procedure requires less time (15 min from sample extraction to extract preparation) and less solvent (approximately one-tenth) compared with conventional methods and is suitable for a fast, economical screen. Additional cleanup procedures, involving dialysis or extraction into base, are described for samples containing high levels of interfering compounds. Thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) with fluorescence detection are described for identification and estimation of mycotoxins. The method has been applied to a wide range of mixed feeds, including laboratory animal diets, and raw materials. The limit of detection is 1 microgram/kg for all mycotoxins measured by HPLC.     

Development and validation of a lateral flow device for the detection of nicarbazin contamination in poultry feeds

Concentrations of the coccidiostat nicarbazin as low as 2 mg/kg in feed can result in violative drug residues arising in poultry liver. A lateral flow device (LFD) was developed for the detection of contaminating concentrations of nicarbazin following solvent extraction of poultry feeds. Test results, as determined by both visual and instrumental measurement, are available within minutes. For 22 feed samples, nicarbazin-free and fortified at 2 mg/kg, the % relative inhibition ranged from 0 to 45% and from 53 to 85%, respectively. Nicarbazin contamination at the critical concentration (2 mg/kg) can be determined in all cases providing the sampling is representative. A wide range of feed samples taken at a mill that incorporated nicarbazin into poultry feed were analyzed. Data generated for these samples by both the LFDs and a mass spectrometric method were compared, and a significant correlation was achieved.     

仔猪配合料比热预测模型的构建

为探究调质、制粒等热处理过程中仔猪配合粉料传热的基本特性,有效控制热量供给并实现调质温度、时间等工艺参数的优化评估,该文以4种粉碎粒度（粉碎筛片孔径：Φ1.0、Φ1.5、Φ2.0和Φ2.5 mm）的仔猪配合粉料为研究对象,采用DSC (differential scanning calorimetry,差示扫描量热法)的方法测定了其在不同含水率范围（10%～18%,湿基）和不同温度范围（25～100℃）的比热,并分析了3个变量对比热的影响规律,构建了比热关于研究变量的预测模型。结果显示：仔猪配合粉料的比热受含水率、温度、粉碎粒度的显著影响,其中含水率的影响最为显著,温度及粉碎粒度次之;比热亦受含水率与粉碎粒度及其与温度的交互作用影响。比热随含水率和温度的增加在1.533～2.881 kJ/(kg·K)范围内升高,且与含水率呈线性关系,与温度呈非线性关系。粉碎过Φ2.5和Φ2.0 mm的仔猪料比热较粉碎过Φ1.5和Φ1.0 mm的大,在高温（60～100℃）和高含水率（18%）时表现尤为明显。基于试验数据建立了比热关于含水率、温度、粉碎粒度的多元回归模型（R2>0.993）,可以有效预测仔猪配合料在此研究变量范围内的比热。研究结果为仔猪配合料调质过程传热方程的建立提供基础数据,并为其热处理工艺参数的优化提供理论依据。     

Microbiological turbidimetric analysis of low chlortetracycline concentrations in feeds.

Recovery studies in which chlortetracycline hydrochloride (CTC-HCI) standard was added to cattle and swine feed supplements at 4.09-9.99 g/ton showed lower antibiotic recovery turbidimetrically (80.6-98.7%) than by the AOAC modified standard as in 38.179(d) (91.2-98.7%) and the plain buffer as in 38.179(b) (93.8-133.0%) methods. Three feeds fortified with a commercial premix at the levels of 5.0 and 10.0 g CTC-HCI/ton showed an overall CTC-HCI recovery of 87.6-110.6% by manual turbidimetric assay. Results were 89.1-108.7% by the AOAC inactivated feed diluent standard and 95.4-125.4% by the plain buffer methods. For some sample extracts (as in cattle feed) the use of heat to stop bacterial growth in the turbidimetric method caused formation of a precipitate. Cooling of cultures to room temperature and rapid reading of sample turbidity followed by standard curve concentrations minimized this interference. The manual turbidimetric assay of low levels of CTC-HCI in feeds appears to offer advantages over other methods.     

肉大鸡颗粒饲料的吸湿解吸平衡模型

为了研究成品饲料的吸湿解吸平衡规律,用GAB模型及其含温度变量的改良模型（mGAB, modified GAB）拟合静态法获得的肉大鸡颗粒料在15、25、35和45℃环境下,在11%至97%平衡相对湿度（即水活度0.11～0.97）范围内的吸湿平衡以及11%～75%（即水活度0.11～0.75）范围内的解吸平衡,得到优异的拟合精度。结果表明：随温度上升,饲料的平衡含水率下降,吸湿和解吸之间存在明显的滞后现象。用BET解吸模型计算的15～45℃范围内单层含水率为6.34%～5.39%（干基）,并且随温度升高线性下降。以水活度0.65为安全贮存的临界点,对应于吸湿和解吸过程, 用改良GAB模型估计4个温度点的安全水分（干基）分别为13.09%和14.71%、12.92%和14.33%、12.74%和14.00%、12.56%和13.66%。研究结果为饲料的贮存和冷却过程规律研究提供了参考。     

Microbiological determination of neomycin in feeds and formulated products

An AOAC modified method is described for the microbiological assay of neomycin, which has been adapted to include complete feeds, supplements, premixes, liquids, oil suspensions, boluses, and antibiotic-impregnated paper. The method features a more sensitive standard response line with a monolayer plating system. The use of a buffered plating medium in place of the water-prepared medium results in a curve with less degree of slope, which allows for more accurate interpretation of the standard response. The feed extract diluent used for standard response line dilution, which is prepared from exposure of the feed extract fluid to pH changes, heat, and sodium hypochlorite, has been eliminated. The constant salt concentration diluent used for the preparation of standards is the same as the salt concentration of the sample extract solution to be tested. Results for 50 commercial complete feeds and 50 commercial premixes received over the last 5 years produced an overall mean recovery of 101% with a mean percent recovery range of 80-112%. A statistical analysis of these 100 commercial, complete feeds and premixes, ranging in concentration from 47 g/ton to 70 g/lb, indicates the assay has little, if any, concentration-related bias. Precision and accuracy of the method was supported by laboratory studies of 20 assays that produced a mean recovery of 101% and standard deviation of 3.     

Applications of hierarchical cluster analysis (CLA) and principal component analysis (PCA) in feed structure and feed molecular chemistry research, using synchrotron-based Fourier transform infrared (FTIR) microspectroscopy

Synchrotron technology based Fourier transform infrared microspectroscopy (S-FTIR) is a recently emerging bioanalytical microprobe capable of exploring the molecular chemistry within microstructures of feed tissues at a cellular or subcellular level. To date there has been very little application of hierarchical cluster analysis (CLA) and principal component analysis (PCA) to the study of feed inherent microstructures and feed molecular chemistry between feeds and/or between different structures within a feed, in relation to feed quality and nutrient availability using S-FTIR. In this paper, multivariate statistical methods--CLA and PCA--were used to analyze synchrotron-based FTIR individual spectra obtained from feed inherent microstructures within intact tissues by using the S-FTIR as a novel approach. The S-FTIR spectral data of three feed inherent structures (strucutre 1, feed pericarp; structure 2, feed aleurone; structure 3, feed endosperm) and different varieties of feeds within cellular dimensions were collected at the National Synchrotron Light Source (NSLS) at Brookhaven National Laboratory (BNL), U.S. Department of Energy (NSLS-BNL, New York). Both PCA and CLA methods gave satisfactory analytical results and are conclusive in showing that they can discriminate and classify inherent structures and molecular chemistry between and among the feed tissues. They also can be used to identify whether differences exist between the varieties. These statistical analyses place synchrotron-based FTIR microspectroscopy at the forefront of those new potential techniques that could be used in rapid, nondestructive, and noninvasive screening of feed intrinsic microstructures and feed molecular chemistry in relation to the quality and nutritive value of feeds.