酶联免疫吸附法快速测定不同样品 基质中三聚氰胺

目的 探求不同样品基质中三聚氰胺残留量的快速检测方法.为快速筛查不同种类食品中非法添加三聚氰胺提供技术保障.方法 采用酶联免疫吸附法在奶制品(奶粉、液态奶)、成品饲料(鸡饲料、猪饲料)、饲料原料(鱼粉、肉骨粉、豆粕、麸皮)、肉类(鸡肉、猪肉、内脏)等样品基质中添加一定浓度的三聚氰胺进行测定,并对检测结果进行分析.结果 酶联免疫试剂盒对奶制品和肉类检出限均能达到1.0 mg/kg;对成品饲料基质中的三聚氰胺的检测,检出限可达到2 mg/kg,而对饲料原料中的三聚氰胺的检测,检出限都不能达到2 mg/kg.结论 对奶制品中的三聚氰胺检测完全符合我国的临时限量标准;对成品饲料中的三聚氰胺残留的检测同样符合其限量标准(2.5 mg/kg),而对饲料原料中的三聚氰胺的检测,由于不同基质中检出限不同,不能直接采用酶联免疫法进行快速筛查;酶联免疫法也适用于肉类中的三聚氰胺的检测.     

Evaluation of a rapid PCR-based method for the detection of animal material

A rapid PCR-based analytical method for detection of animal-derived materials in complete feed was developed. Using a commercially available DNA forensic kit for the extraction of DNA from animal feed, a sensitive method was developed that was capable of detecting as little as 0.03% bovine meat and bone meal in complete feed in under 8 h of total assay time. The reduction in assay time was accomplished by reducing the DNA extraction time to 2 h and using the simpler cleanup procedure of the kit. Assay sensitivity can be increased to 0.006% by increasing the DNA extraction time to an overnight incubation of approximately 16 h. Examination of dairy feed samples containing either bovine meat and bone meal, porcine meat and bone meal, or lamb meal at a level of 0.1% (wt/wt basis) suggested that this method may be suitable for regulatory uses. The adoption of this commercially available kit for use with animal feeds yields an assay that is quicker and simpler to perform than a previously validated assay for the detection of animal proteins in animal feed.     

Supplying the protein needs of dairy cattle from by-product feeds

Several by-product feeds are relatively high in crude protein and exhibit relatively low ruminal degradability, which make them desirable proteinaceous feeds for dairy cows. Therefore, by-product feeds have been and will continue to be important feeds for dairy cows. Factors are discussed that affect ruminal degradability of protein in distillers grains, distillers grains with solubles, brewers grains, corn gluten feed, corn gluten meal, meat meal, meat and bone meal, blood meal, and fish meal, and the potential of these feeds to provide supplemental amino acids needed by lactating dairy cows. The importance of maximizing synthesis of microbial protein and digestion of organic matter in the rumen is emphasized in relation to total amino acid passage to the small intestine. For these feeds to be used most successfully, they must be available from a dependable source at an economical cost and should supply amino acids that complement other amino acids passing to the small intestine. Benefits that should be realized from the successful use of by-product feeds include increased milk production from feeding proteins that have greater ruminal escape potentials and a reduced cost per unit of milk produced because of decreased use of expensive supplemental protein.     

菜籽粕替代鱼粉对银鲫生长性能及饲料利用率的影响

为评估硬颗粒饲料和膨化饲料中菜籽粕替代鱼粉对银鲫生长及饲料利用率的影响,试验设计了4种等氮、等能的饲料,分别用菜籽粕替代饲料中0％(对照组)、25％、50％、75％的鱼粉,利用挤压膨化技术和环模制粒技术加工成4种膨化饲料和4种硬颗粒饲料,对银鲫进行为期50 d的饲养试验.结果显示:菜籽粕替代鱼粉饲喂银鲫后的生长性能及饲料利用率都呈现下降的趋势;相同饲料配方,饲喂膨化饲料的银鲫生长性能及饲料利用率优于饲喂硬颗粒饲料的银鲫.研究结果表明,菜籽粕替代银鲫饲料中适量的鱼粉是可行的,膨化饲料饲喂银鲫的效果优于硬颗粒饲料,利用膨化加工工艺可以显著提高银鲫饲料中菜籽粕对鱼粉替代量.     

Total and ileal digestible tryptophan contents of feedstuffs for broiler chickens

Reliable values of total and digestible tryptophan in components of feed formulation matrices are needed because tryptophan is often the third limiting amino acid in practical poultry diets. However, tryptophan is oxidatively destroyed during acid hydrolysis in routine amino acid analysis and its determination requires a separate analytical procedure. The variability in contents and apparent ileal digestibility for 6‐week‐old broiler chickens of tryptophan in 74 samples representing 24 feedstuffs are presented in this paper. The average ileal tryptophan digestibility coefficient in wheat was 0.83, in sorghum and triticale 0.75, maize 0.71, soybean meal 0.84, sunflower meal 0.81, canola meal 0.78 and cottonseed meal 0.75. Among the grain legumes, tryptophan in lupins was better digested than that in chickpeas, fababeans and field peas. Among the animal protein meals, the tryptophan digestibility coefficients in fish meal (0.77) and blood meal (0.84) were substantially higher than those in meat meal (0.62), meat‐and‐bone meal (0.63) and feather meal (0.52). Marked variations in tryptophan digestibility were also observed among samples of fish meal, meat‐and‐bone meal and meat meal, highlighting significant batch‐to‐batch differences. For most feedstuffs, considerable variability was observed in the tryptophan concentrations, but such variations were not reflected in digestibility coefficients. Copyright © 2006 Society of Chemical Industry     

利用可视基因膜芯片技术与实时荧光定量聚合酶链式反应技术分析肉制品中动物源性成分

为调查了解肉制品中动物源性成分,以帮助判别掺假情况,应用可视基因膜芯片检测技术对市售的肉松、香肠、肉卷、预制调理肉、肉干及肉脯等23份样品动物源性成分进行筛查分析,同时,针对筛查结果采用实时荧光定量聚合酶链式反应（polymerase chain reaction,PCR）法进一步确证。结果表明：可视基因膜芯片检测法与实时荧光定量PCR法检测结果一致,提高了未知样品的筛查效率;在本次随机分析的样品中,动物源性成分检测结果与标签标示不一致的情况占比高达21.7%,肉制品掺假虚标情况不容忽视。     

Evaluation of two commercial lateral-flow test kits for detection of animal proteins in animal feed

Performance characteristics were evaluated for two lateral-flow test kits, Reveal for Ruminant in Feed (Neogen Corporation) and FeedChek (Strategic Diagnostics Inc.), designed to detect ruminant or terrestrial animal proteins in feeds. The stringent acceptance criteria used were developed by the Center for Veterinary Medicine Office of Research to identify test kits with comparable selectivity and sensitivity to microscopy and PCR assay, the analytical methods used by the U.S. Food and Drug Administration (FDA). Guidelines were developed for evaluating the selectivity, sensitivity, ruggedness, and specificity of these kits. These guidelines further stated that ruggedness and specificity testing would be performed only after a test passed both the selectivity and sensitivity assessments. Acceptance criteria for determining success were developed using a statistical approach requiring 90% probability of achieving the correct response, within a 95% confidence interval. A minimum detection level of 0.1% bovine meat and bone meal, consistent with the sensitivity of the methods used by the FDA, was required. Selectivity was assessed by testing 60 dairy feed samples that contained no added animal proteins; sensitivity was determined by evaluating 60 samples (per level of fortification) of the same feed that contained 0.025, 0.05, 0.1, 0.25, 0.5, 1, or 2% bovine meat and bone meal. The Reveal test passed the selectivity assessment but failed the sensitivity assessment, detecting only samples fortified at the 2% level and then only 17 to 33% of those samples, when read according to the label directions. The FeedChek test passed the sensitivity assessment but failed the selectivity assessment, with rates for false-positive results ranging from 34 to 38%, depending on the user. The sensitivity of the Reveal test was affected by the concentration of trace minerals present in the feed; concentrations toward the high end of the normal range prevented the detection of true positive feed samples containing bovine meat and bone meal. Better sensitivity assessments were obtained when lamb meal was used either alone or in combination with bovine meat and bone meal. The FeedChek test was not affected by the concentration of trace minerals or by the type of animal meal used. These results indicate that neither of the two tests is adequate for routine regulatory use.     

Monoclonal antibody-based sandwich enzyme-linked immunosorbent assay for sensitive detection of prohibited ruminant proteins in feedstuffs

Regulations aimed to control the epidemic of bovine spongiform encephalopathy have banned the use of certain animal products, i.e., ruminant meat and bone meals, in ruminant animal feeds. A sensitive enzyme-linked immunosorbent assay has been developed to detect prohibited bovine and ovine muscles in feedstuffs. The assay utilizes a pair of monoclonal antibodies (MAbs) against skeletal troponin I (TnI). MAb 5G9, specific to bovine and ovine TnI, was used as the capture antibody and the biotin-conjugated MAb 2G3, reacting to all heterologous TnI, was used as the detection antibody. Quantitative procedures were applied to samples containing 5, 0.5, and 0.05% (wt/wt) of heat-treated (132 degrees C/2 bar, 2 h) bovine and ovine meat meals in three different feeds, coexisting with porcine, chicken, or turkey meat meal. The presence of these nonprohibited species did not affect the detection of bovine and ovine meat meals in the feed samples (P > 0.05). Quantitative determinations of extractable bovine and ovine TnI, with a detection limit of 5.0 and 4.0 ng/ml, respectively, were achieved when the matching feed matrixes were used in the calibration curves. This new assay provides a rapid and reliable way to detect animal protein products containing a trace amount of bovine or ovine muscle tissue in feedstuffs.     

Effect on protein digestibility of different processing conditions in the production of fish meal and fish feed

The effect of processing conditions on protein digestibility and fluorodinitrobenzene (FDNB)‐reactive (available) lysine in the production of fish meal and extruded fish feed has been studied under pilot and commercial conditions using mink as model animals. Fish meal produced under pilot‐plant conditions at processing temperatures below 70–80 °C (FM1) had protein digestibility of 929 (grams of protein digested per 1000 g protein consumed) compared with 905 when processed at temperatures above 100 °C (FM2). A low‐temperature‐processed commercial fish meal (CFM1) had protein digestibility of 940 compared with 888 for a standard commercial fish meal (CFM2). Pilot‐produced extruded fish feed had protein digestibility of 913 when based on FM1 as the main protein source (95% of total protein) compared with 892 when based on FM2. Commercial extruded fish feed had protein digestibility of 912 when based on CFM1 compared with 871 when based on CFM2. Varying extrusion conditions at the pilot scale, ie temperatures from 100 to 126 °C and moisture contents from 21 to 12%, did not affect protein digestibility. Similarly, under commercial conditions, variation in temperature from 89 to 110 °C and moisture from 24.5 to 19.5% did not affect FDNB‐reactive lysine and protein digestibility. The FDNB‐reactive lysine content and protein digestibility of the extruded feed were less than the values calculated from the ingredient mixture before extrusion. Thus, despite different extrusion conditions not giving different FDNB‐reactive lysine and protein digestibility, the total process, ie extrusion, drying and oil coating, caused a reduction. Copyright © 2003 Society of Chemical Industry     

Nutritional evaluation of seed proteins of Actinodaphne hookeri

The possibility of utilisation of pisa meal, which remains after defatting of Actinodaphne hookeri seeds, as a potential protein source in foods and feeds has been investigated. Rat growth experiments with half of the dietary protein derived from the meal were carried out, using casein as standard protein for comparison. Reasonably good correlation between chemical data and biological performance was observed. The meal, when made nutritionally adequate by supplementation with methionine and lysine, was capable of supporting normal growth and maintaining other physiological functions in rats. Feeding the rats with pisa meal for 8-10 weeks did not reveal any abnormal biochemical lesion indicative of the onset of toxic action.     

Apparent ileal digestibility of amino acids in feed ingredients determined with broilers and layers

The apparent ileal digestibility of amino acids in eight feed ingredients were determined using broilers and layers. The ingredients included three cereals (wheat, sorghum and maize), one cereal by‐product (wheat middlings), three oilseed meals (canola, cottonseed and soybean meals) and one animal protein meal (meat and bone meal). Dietary protein in the assay diets was supplied solely by the test ingredient. All diets contained 20 g kg^?1 acid‐insoluble ash as an indigestible marker, and each diet was offered ad libitum in mash form to five replicate pens of 42‐day‐old broilers and 60‐week‐old layers. The digestibility coefficients of most amino acids for wheat and sorghum were similar (P > 0.05) in broilers and layers. The digestibility of most amino acids for maize was higher (P < 0.05) in broilers compared to those in layers. The digestibility of individual amino acid for wheat middlings was higher (P < 0.05) in layers than in broilers. In general, the digestibility of amino acids for cottonseed meal, soybean meal, and meat and bone meal were similar (P > 0.05) between broilers and layers. The influence of class of bird on digestibility in canola meal was variable. The digestibility of threonine, valine, isoleucine, leucine, phenylalanine, glutamic acid and alanine were higher (P < 0.05), and those of methionine, histidine and lysine were lower (P < 0.05) in broilers compared to layers. These results suggest that the practice of using amino acid digestibility values generated with broilers for layers may not be appropriate for all feed ingredients. Copyright © 2006 Society of Chemical Industry     

Bioaccessibility,cellular uptake and transepithelial transport of α‐tocopherol and retinol from a range of supplemented foodstuffs assessed using the caco‐2 cell model

The bioaccessibility, or amount of a nutrient available for gastrointestinal absorption, can be determined using an in vitro digestion model, the addition of the resultant digestate to a caco‐2 transwell model system yields an approximation of nutrient bioavailability. The objective of the present study was to compare the bioaccessibility and bioavailability of α‐tocopherol and retinol from a range of digested foodstuffs. Minced pork, beef and turkey and apple sauce, bread and mayonnaise were supplemented with α‐tocopherol‐acetate and retinol‐acetate prior to being subjected to an in vitro digestion procedure. The aqueous fraction of each of the digested foodstuffs was then added to a caco‐2 transwell model and the transepithelial transport was determined. The findings of the present study indicate that α‐tocopherol and retinol are more bioaccessible from supplemented meat products than from supplemented apple sauce, bread and mayonnaise. It was found that turkey meat facilitated the highest bioaccessibility and subsequent cellular uptake and transport of retinol. The cellular uptake and secretion of α‐tocopherol was similar for all samples.     

Evaluation of Laboratory Techniques for Predicting Ruminal Protein Degradation

Five methods of measuring protein solubility, an in situ method, and an in vitro method for measuring protein degradability were evaluated to determine which procedure most accurately predicted quantity of feed protein escaping ruminal fermentation. Feeds evaluated were soybean meal, blood meal, meat meal, corn gluten meal, distillers dried grains, brewers dried grains, distillers grains plus solubles, dehydrated alfalfa, and soybean meal treated with sodium bentonite. Solubility was measured in .15 M sodium chloride, 10% Burroughs solution, .02 N sodium hydroxide, hot water, and bicarbonate-phosphate buffer. In situ procedure was the incubation of feeds in dacron bags suspended in the rumen of cattle. In vitro procedure was incubation of feeds with five proteolytic enzymes. Results from these methods were correlated with protein degradability determined in vivo or calculated from growth trials. Protein solubility in hot water, 10% Burroughs solution, and bicarbonate-phosphate buffer was closely correlated with in vivo protein degradability, .86, .69, and .87. Highest correlation for dacron bag incubations with in vivo degradability were at 12 and 24 h, .88 and .84. All proteolytic enzymes yielded highly significant correlations with protein degradability with incubations of 1 and 4 h and offer procedural advantages over the dacron bag technique.     

ANTIMICROBIAL AND ANTIOXIDANT ACTIVITIES OF CORN GLUTEN MEAL SUBSTANCES IN AN EMULSIFIED MEAT MODEL SYSTEM

Corn gluten meal (CGM) and pH adjusted (6.6)/particle size-reduced (∼7μm-MC7; ∼38μm-MC38) CGM increased oxidative and microbiological shelf-life of a model emulsified meat system. After 14 days, TBARS were significantly lower for samples containing MC7 and MC38 when compared to controls, and samples containing soy protein isolate (SPI) or native CGM. Based on aerobic plate counts, MC38 and CGM increased product shelf-life by ∼7days compared to SPI- and MC7-containing products and controls. These results suggest that the use of corn gluten meal substances in an emulsified meat product could create a more shelf stable product. These functional characteristics may make the incorporation of corn gluten meal into foods more feasible.     

Effect of Sources of Fibre on Performance of Growing Snail

The proximate and physicochemical properties of cassava leaf and peel meals were evaluated with a view to possible replacement of wheat offal which is the conventional source of fibre in animal feed, with these meals. The effect of feeds produced with cassava leaf and peel meals on the performance of growing snails was also investigated. Feeds (F1, F2 and F3) were formulated to contain 240, 235 and 230 g/kg cassava root meal each and 85, 85 and 90 g/kg cassava peel meal, wheat offal meal and cassava leaf meal respectively. The formulated feeds contain approximately 18.0% crude protein, 7.5% ash, 3% fat, 6.0% crude fibre 8%, calcium, 0.7% phosphorus, and energy level of 2400 kcal ME / kg. A total of 54 growing snails (Archachatina marginata) were used to investigate the nutritive potential of the formulated feeds on performance of growing snails for 15 weeks. Concentrations of the crude protein, crude fat, crude fibre, ash and calcium in cassava leaf meal were higher than those of wheat offal and cassava peel meal, with the exception of nitrogen free extract which was highest (70.01%) in cassava peel meal. Feed intake was 576 g 569 g and 581 g for snails fed with cassava leaf meal, cassava peel meal and wheat offal respectively but the corresponding weight gain ranged between 123.35 and 134.81 % being highest for F1. The feed conversion ratio shows that F1 > F3 > F2 indicating better conversion of feed to edible meat in F1. The results show that cassava leaves and peels have a strong potential to substitute the traditional wheat offal and can therefore be adapted as commercial feed ingredients.     

In-vitro determination of nitrogen digestibility and lysine availability in meat and bone meals and comparison with in-vivo ileal digestibility estimates

The true ileal digestibility of nitrogen was determined in 20 meat and bone meal samples using 120 growing rats. Correction for endogenous ileal nitrogen excretion was based on feeding a further 15 rats a protein-free diet. The in-vivo digestibility estimates were compared with in-vitro values determined using either a multi-enzyme digestibility assay (method A), the pronase assay (method B) or a multi-enzyme digestion, pH-drop assay (method C). The in-vivo true digestibilities of lysine for 12 of the meat and bone meal samples were compared with fluoro-dinitrobenzene (FDNB) available lysine values. The mean in-vivo digestibility values ranged from 57.2 to 79.1 % and the mean coefficient of variation between rats within meals was 7.6%. For methods A and B the correlations between in-vitro and in-vivo digestibility were low and not significant (P>0.05). There was a significant positive correlation between pH at 10 min from the start of in-vitro digestion and in-vivo digestibility (r= +0.75; P <0.001) as was the case for pH at 20 min (r= +0.52; P <0.05). The positive relationship between pH at 10 or 20 min and in-vivo digestibility was contrary to expectation and may have been caused by the potentially high buffering capacity of meat and bone meal. The in-vitro methods A, B and C each showed a high degree of precision. The mean in-vivo true ileal digestibility of lysine in the meat and bone meals ranged from 64.7 to 86.9 %. There was no significant correlation between in-vivo lysine digestibility and FDNB available lysine. There appeared to be an interaction between FDNB lysine availability and true ileal lysine digestibility, such that the meals with lower in-vivo digestibility had availability values higher than the corresponding in-vivo digestibility values.     

Development of primers for detection of meat and bone meal in ruminant feed and identification of the animal of origin

The safe use of cattle feed free from meat and bone meal is an important prerequisite to prevent further spread of bovine spongiform encephalopathy. We designed primers to detect very small amounts of meat and bone meal in ruminant feed. Mitochondrial subunit 8 of the ATP synthase gene was used as a target sequence. PCR-based assays revealed amplification of DNA from mammals, ruminants, and individual species using these primers. The method allowed detection of the presence of meat and bone meal in ruminant feed from 0.1 to 0.01%. Sensitivity and effectiveness of the method for detecting prohibited animal proteins in ruminant feed was evaluated.     

The choline content of feed ingredients and mixed feeds determined by an enzymatic assay

A recently developed procedure for the determination of choline in ingredients and feeds was utilised to establish the levels and the variability of choline in a series of ingredients and to compare the analysed and calculated choline values of mixed feeds. The concentrations of choline in samples of maize (7), canola meal (3), wheat middlings (1) and dehydrated bakery product (1) were, respectively, 1·55±0·18, 7·59±0·08, 2·35 and 2·39 g kg⁻¹, all higher than ingredient composition table values. Choline contents lower than table values were found in samples of poultry by-products (7) and meat and bone meal (6): 2·18±0·87 and 1·08±0·29 g kg⁻¹, respectively. The average choline concentration found in samples of dehulled soybean meal (7) was 2·73±0·18 g kg⁻¹, similar to table values. The choline in samples of poultry fat (2) averaged 0·48±0·02 g kg⁻¹. Significant correlations between the concentrations of choline and of some components of the proximate analysis were found. The analysed choline concentrations in mixed feeds were only 1·4% lower than the calculated levels based on the ingredient analyses. The procedure was adequate for choline determination in ingredients or mixed feeds. The high variability in the choline content of some ingredients may require analysis for proper feed formulation. © 1998 Society of Chemical Industry.     

Production responses of dairy cows fed various amounts of rumen-protected methionine and lysine

A 3 x 3 factorial response surface design was used to study the effects of feeding rumen-protected methionine and lysine to dairy cows between 22 and 305 d of lactation. A total of 130 dairy cows at three universities were individually fed a corn silage and corn grain-based diet that contained either soybean meal or corn gluten meal and urea. An unsupplemented control diet plus nine treatment combinations of three amounts of rumen-protected DL-methionine (3.4, 7.8, and 12.2 g/d) and three amounts of rumen-protected L-lysine (5.9, 13.5, and 21.1 g/d) were fed at all locations. Plasma concentrations of methionine and lysine were increased when rumen-protected methionine and lysine were supplemented to the diets. Rumen-protected methionine and lysine did not affect feed intake by cows fed either by soybean meal or corn gluten meal and urea based diets. Milk protein percentage was increased, but milk and milk protein yields were not improved when diets containing soybean meal were supplemented with rumen-protected methionine and lysine. In contrast, milk and milk protein yields were improved when a diet that contained corn gluten meal and urea was supplemented with rumen-protected methionine and lysine. Health and reproduction measurements were similar for cows receiving all treatments.     

Validation of a PCR-based method for the detection of various rendered materials in feedstuffs using a forensic DNA extraction kit

A method trial was initiated to validate the use of a commercial DNA forensic kit to extract DNA from animal feed as part of a PCR-based method. Four different PCR primer pairs (one bovine pair, one porcine pair, one ovine primer pair, and one multispecies pair) were also evaluated. Each laboratory was required to analyze a total of 120 dairy feed samples either not fortified (control, true negative) or fortified with bovine meat and bone meal, porcine meat and bone meal (PMBM), or lamb meal. Feeds were fortified with the animal meals at a concentration of 0.1% (wt/wt). Ten laboratories participated in this trial, and each laboratory was required to evaluate two different primer pairs, i.e., each PCR primer pair was evaluated by five different laboratories. The method was considered to be validated for a given animal source when three or more laboratories achieved at least 97% accuracy (29 correct of 30 samples for 96.7% accuracy, rounded up to 97%) in detecting the fortified samples for that source. Using this criterion, the method was validated for the bovine primer because three laboratories met the criterion, with an average accuracy of 98.9%. The average false-positive rate was 3.0% in these laboratories. A fourth laboratory was 80% accurate in identifying the samples fortified with bovine meat and bone meal. A fifth laboratory was not able to consistently extract the DNA from the feed samples and did not achieve the criterion for accuracy for either the bovine or multispecies PCR primers. For the porcine primers, the method was validated, with four laboratories meeting the criterion for accuracy with an average accuracy of 99.2%. The fifth laboratory had a 93.3% accuracy outcome for the porcine primer. Collectively, these five laboratories had a 1.3% false-positive rate for the porcine primer. No laboratory was able to meet the criterion for accuracy with the ovine primers, most likely because of problems with the synthesis of the primer pair; none of the positive control DNA samples could be detected with the ovine primers. The multispecies primer pair was validated in three laboratories for use with bovine meat and bone meal and lamb meal but not with PMBM. The three laboratories had an average accuracy of 98.9% for bovine meat and bone meal, 97.8% for lamb meal, and 63.3% for PMBM. When examined on an individual laboratory basis, one of these four laboratories could not identify a single feed sample containing PMBM by using the multispecies primer, whereas the other laboratory identified only one PMBM-fortified sample, suggesting that the limit of detection for PMBM with this primer pair is around 0.1% (wt/wt). The results of this study demonstrated that the DNA forensic kit can be used to extract DNA from animal feed, which can then be used for PCR analysis to detect animal-derived protein present in the feed sample.