Yun-Hwa Peggy Hsieh

Detection of horse meat contamination in raw and heat-processed meat products.

Europes recent problems with the adulteration of beef products with horse meat highlight the need for a reliable method for detecting horse meat in food for human consumption. The objective of this study was therefore to develop a reliable monoclonal antibody (mAb) based enzyme-linked immunosorbent assay (ELISA) for horse meat detection. Two mAbs, H3E3 (IgG2b) and H4E7 (IgG2a), were characterized as horse-selective, and competitive ELISAs (cELISAs) employing these mAbs were developed. The cELISAs were found to be capable of detecting levels as low as 1% of horse meat in raw, cooked, and autoclaved ground beef or pork, being useful analytical tools for addressing the health, economic, and ethical concerns associated with adulterating meat products with horse meat. However, due to cross-reaction with raw poultry meat, it is recommended that samples be heated (100 °C for 15 min) prior to analysis to eliminate possible false-positive results.

Issues related to the use of blood in food and animal feed.

Blood has traditionally been used as a high protein ingredient in both human food and animal feed, with resulting economic, environmental and nutritional benefits. However, potentially serious health and safety issues related to blood consumption, particularly the risk of pathogenic or harmful metabolic materials, the infectivity of prion diseases, and the presence of identified allergens such as bovine serum albumin (BSA), are causing many consumers to shy away from any product containing either animal blood or ingredients derived from animal blood. Thus, despite the significant volumes of blood produced by slaughterhouses, blood is currently underutilized as a food ingredient. This article reviews the use of animal blood as an ingredient in food intended for human consumption or for animal feed and discusses the related consumer concerns.

Characterization of a 12 kDa Thermal‐Stable Antigenic Protein in Bovine Blood

UNLABELLED We have previously developed an immunoassay based on monoclonal antibody (MAb) Bb1H9 for quantitative detection of ruminant blood in processed food and feedstuffs. The purpose of this study was to characterize the unknown 12 kDa thermal-stable ruminant-specific antigenic protein recognized by MAb Bb1H9 in order to better define the application scope of the developed assay. Extracts obtained from raw and heat-treated bovine blood-derived products were analyzed with indirect ELISA and Western blot. Target proteins resolved by 2D electrophoresis were subjected to N-terminal sequencing. Results indicated that the 12 kDa protein is a monomer of the tetrameric hemoglobin molecule (64.5 kDa) and that the heme group is not required for its binding with MAb Bb1H9. This MAb can be utilized as a probe for red blood cell derived products of ruminant origin in raw or processed food and feedstuffs to enforce labeling regulations and to address consumer concerns. PRACTICAL APPLICATION MAb Bb1H9 represents the first antibody with the capacity to recognize bovine hemoglobin both in the absence and presence of the heme group, regardless of the heat treatment. MAb Bb1H9 can therefore be utilized in immunoassays by manufacturers and regulators to detect any ingredients containing hemoglobin or globin (hemoglobin without the heme group) in both raw and processed food and feed materials for product quality control and labeling law enforcement.

Monoclonal Antibodies as Probes for the Detection of Porcine Blood-Derived Food Ingredients

The lack of effective methods to monitor the use of porcine blood-derived food ingredients (PBFIs) is a concern for the billions of individuals who avoid consuming blood. We therefore sought to develop a panel of porcine blood-specific monoclonal antibodies (mAbs) for use as probes in immunoassays for the detection of PBFIs. Ten selected mAbs were identified that react with either a 60 or 90 kDa protein in the plasma fraction or a 12 kDa protein in the red blood cell fraction of porcine blood. Western blot analysis of commercially produced PBFIs revealed that these antigenic proteins are not affected by various manufacturing processes. The utility of these mAbs was demonstrated in a prototype sandwich ELISA developed for this study using mAbs 19C5-E10 and 16F9-C11. The new assay is porcine blood-specific and capable of detecting ≤0.03% (v/v) of PBFIs in cooked (100 °C for 15 min) ground meats or fish.

Characterization of a 60-kDa Thermally Stable Antigenic Protein as a Marker for the Immunodetection of Bovine Plasma-Derived Food Ingredients.

A sandwich enzyme-linked immunosorbent assay (sELISA) based on 2 monoclonal antibodies (Bb3D6 and Bb6G12) that recognize a 60-kDa antigenic protein in bovine blood was previously developed for detecting bovine blood in animal feed for the prevention of mad cow disease. This study sought to establish the identity of this 60-kDa antigenic protein and consequently determine the suitability of the sELISA for detecting bovine plasma-derived food ingredients (BPFIs), which are widely used in dietary products without explicit labeling. Results from western blot confirmed the 60-kDa protein to be present in the plasma fraction of bovine blood. Further proteomic analyses involving 2-dimensional gel electrophoresis (2-D GE) and amino acid sequencing revealed the 60-kDa protein to be bovine serum albumin (BSA). The sELISA proved capable of detecting BPFIs in all the commercial dietary supplements tested, including those that were formulated with hydrolyzed BPFIs. The assay could also detect 0.01% and 0.5% of different BPFIs in spiked raw and cooked ground beef, respectively. This assay based on the detection of BSA therefore has the potential to become a valuable analytical tool to protect consumers who avoid consuming BPFIs for religious, health, or ethical reasons.

Speciation of Animal Fat: Needs and Challenges

ABSTRACT The use of pork fat is a concern for Muslims and Jews, who for religious reasons avoid consuming anything that is pig-derived. The use of bovine materials, including beef fat, is prohibited in Hinduism and may also pose a risk of carrying the infectious agent for bovine spongiform encephalopathy. Vegetable oils are sometimes adulterated with animal fat or pork fat with beef fat for economic gain. The development of methods to determine the species origin of fat has therefore become a priority due to the complex and global nature of the food trade, which creates opportunities for the fraudulent use of these animal fats as food ingredients. However, determining the species origin of fats in processed foods or composite blends is an arduous task as the adulterant has a composition that is very similar to that of the original fat or oil. This review examines some of the methods that have been developed for fat speciation, including both fat-based and DNA-based methods, their shortcomings, and the need for additional alternatives. Protein-based methods, specifically immunoassays targeting residual proteins in adipose tissue, that are being explored by researchers as a new tool for fat speciation will also be discussed.