Novel protein sources (like insects, algae, duckweed, and rapeseed) are expected to enter the European feed and food market as replacers for animal-derived proteins. However, food safety aspects of these novel protein sources are not well-known. The aim of this article is to review the state of the art on the safety of major novel protein sources for feed and food production, in particular insects, algae (microalgae and seaweed), duckweed, and rapeseed. Potential hazards for these protein sources are described and EU legislative requirements as regard to food and feed safety are explained. Potential hazards may include a range of contaminants, like heavy metals, mycotoxins, pesticide residues, as well as pathogens. Some safety aspects of novel protein sources are intrinsic to the product, but many potential hazards can also be due to production methods and processing conditions. These aspects should be considered in advance during product development. European law is unclear on several issues regarding the use of novel protein sources in food and feed products. For food product applications, the most important question for food producers is whether or not the product is considered a novel food. One of the major unclarities for feed applications is whether or not products with insects are considered animal-derived products or not. Due to the unclarities in European law, it is not always clear which Regulation and maximum levels for contaminants apply. For market introduction, European legislation should be adjusted and clarified.
In response to the Kyoto Protocol on the global climate, the European Union (EU) made agreements on reducing greenhouse gas emissions. Major sources of these gas emissions are cattle breeding and related meat consumption (Steinfeld and others 2006; Van Beukering and others 2008).
In the last decades, the global consumption of animal proteins has increased continuously. Between 1950 and 2000, the global human population more than doubled from 2.7 to 6 billion people. Meat production, however, increased by a factor of 5 from 45 to 233 billion kg per year. For 2050, Food and Agriculture Organization of the United Nations (FAO) expects a world population of 9 billion and meat production of 410 billion kg per year (Boland and others 2013; FAO 2006). Animal-derived protein accounts currently for almost 40% of humanity's total protein consumption, and FAO expects a substantial increase by 2050 if the trend goes on uninterrupted (Boland and others 2013). Several global developments, such as the increasing demand on animal products in China and India, necessitates to use animal proteins in a more responsible manner than currently (Vereijken and Aiking 2006).
Animal proteins are produced inefficiently. Depending on the animal species and various conditions, 2 to 15 kg plant material is needed to produce 1 kg of animal products. Currently, 40% to 50% of the global grain harvest is used for feed production (Profetas 2008). To reduce the use of area and energy, people should eat less beef and more pork or chicken, reduce their portions, or change the meat portion in the diet by meat substitutes, such as legumes or eggs (Gerbens-Leenes 2000; Aiking and De Boer 2006). Other studies also concluded that reduction of both meat consumption and production will reduce greenhouse gas emissions (Stehfest and others 2008; Van Beukering and others 2008).
Novel protein sources (like insects, algae, duckweed, and rapeseed) are expected to enter the European feed and food market. For instance, the Dutch Ministry of Economic Affairs (EZ) supports, by the Acceleration agenda “Protein innovations,” various possibilities to make current production and consumption of proteins more sustainable by modifying the production of animal proteins, development of new (plant) protein products, and biorefinery and valorization of protein-rich waste streams (Bleker 2012).
Currently, research on novel proteins is conducted on the functionality, processing, and industrial application of novel proteins, and the valorization of by-products or waste streams obtained in other processes. However, food safety aspects of these novel proteins are not well-known. Especially the use of waste streams may introduce multiple food safety hazards. The presence of contaminants, antinutritional factors (ANFs), allergens, and accumulation and modification of substances in protein matrices during processing can have effects on public health.
The objective of this article is to review the state of the art on the safety of major novel protein sources for feed and food production, and on EU requirements for their safety. Unclarities to fulfill safety requirements for the introduction of novel proteins on the EU market are elucidated. First, novel protein sources will be described, as well as the potential hazards for proteins from insects, algae (microalgae and seaweed), duckweed, and rapeseed. Second, the EU requirements for novel protein sources will be explained in general and specifically for the selected protein sources.
Major animal protein sources for human consumption include meat, fish, milk, and eggs. Developments in the production of animal protein sources are directed toward the use of cheaper raw materials, increased health characteristics, and higher sustainability by using existing and novel protein sources and production methods. For this purpose, the use of novel protein sources, waste streams of processing, and by-products of biofuel production are investigated (Krijne and Essink 2011). Various novel protein sources can be used for replacing current animal proteins in feed and food production (see Figure 1), but with different estimated time to enter the protein sources on the market.
Current plant protein sources that are widely consumed are proteins derived from soy, wheat, vegetables, and potatoes. Soy is the most important protein source for the production of animal proteins. Wheat is the largest group of plant protein sources in the Western diet and is often used for feed products (Krijne and Essink 2011).
Legumes (lupine), grains (rice, maize), mushrooms, and potatoes are sources that have already been used in food and feed. The availability and functionality of their proteins will be further improved by new technologies and valorization of waste streams (Stegeman and others 2010; Krijne and Essink 2011).
Another group of protein sources that is currently used for feed and biofuel products includes rapeseed, algae, grass, duckweed, and by-products from agricultural processing and waste streams from biofuel chains (Stegeman and others 2010). Rapeseed is more or less ready now for market introduction for food use (Stegeman and others 2010; Krijne and Essink 2011). The other protein sources mentioned are still in early development stages. The same applies to insect proteins that are now used for petfood and (fish) feed. New processing technologies (such as extraction procedures) and valorization of waste streams, in order to apply insect proteins in feed and food, are being developed (Stegeman and others 2010; Krijne and Essink 2011). Processing insect cells opens up new possibilities for the application of this novel protein source (Verkerk and others 2007). The major novel protein sources are detailed below.
Insects consist of many species. Potential insects for application in food in the EU are Gryllodus sigillatus and Acheta domesticus (crickets), Alphitobius diaperinus (lesser mealworm), and Tenebrio molitor (yellow mealworm). Potential insects for use in feed products in the EU are Hermetia illucens (black soldier fly), larvae of Musca domestica (common housefly), and T. molitor (yellow mealworm).
During the rearing of insects, a metamorphosis occurs from egg via larva and pupa to mature adult. The modification path varies depending on the subspecies. Feed used for these insects are, for example, chicken feed, vegetables, and waste streams. Whole insects can be eaten raw, dried, crushed, textured, pulverized, or ground, or they are heated (such as cooked, boiled, fried, roasted, toasted, extruded, and canned) or preserved by freeze-drying or cryovacking after degutting or fasting. Insect proteins can also be isolated by extraction before use in food products.
Most edible insects in non-European countries are harvested from the wild (DeFoliart 1995). According to Schabel (2008), some insects may be considered as edible in some regions but not in others; some normally safe insects may be unhealthy if they feed on certain plants or originate from a polluted and pesticide-treated area; some may be safe for some consumers but less (or not at all) for others due to the presence of allergens; and some may require special capture, preparation, storage, or transportation methods to render and keep them safe (Schabel 2008).
The long history of human consumption of insects in non-EU countries suggests, with little evidence to the contrary, that insects harvested for human consumption do not cause any significant health problems (DeFoliart 1992). Zhou and Han (2006) describe a safety evaluation of silkworm by a series of acute and subacute toxicological tests. Their results indicate that protein of silkworm pupae can be generally regarded as safe at a maximum dose of 1.50 g/kg body weight per day in rats (Zhou and Han 2006). Sirimungkararat and others (2008) reported that consumption of processed eri products derived from eri silkmoths was safe in terms of the presence of these toxic chemical substances: hydrocyanic acid, heavy metals (lead, mercury, and cadmium), arsenic, benzoic acid, and sorbic acid. However, there are no standards for eri foods yet, and allowable levels were used that are defined by the Thai Community Product Standard (TCPS) for other (nonreported) products (Sirimungkararat and others 2008). The U.S. Food and Drug Administration (FDA) has prescribed permissible levels of contamination of food with insect debris (FDA 2011). People are regularly eating small amounts of insects unconsciously, but no serious complications have been observed, with the exception of individuals who react allergically (Mitsuhashi 2008).
In 2010, the Codex Alimentarius Commission (CAC) reported that food safety of edible insects has not been studied extensively, which may be due to the fact that these insects are often treated as traditional foods of indigenous populations and rarely recognized as tradable food items. CAC describes that insects are rich in nutrients providing a medium for growth of unwanted microorganisms under certain conditions, especially making uncooked insects susceptible for microbiological hazards unless proper heat treatment or storage conditions are applied. Some people have or may develop (food) allergies against certain species of insects. Some insects may require other treatments before they are rendered edible (CAC 2010). Insect proteins may display a cross-allergenicity with shrimps and house dust mites (Witteman and others 1994; Leung and others 1996; Reese and others 1999; Houben 2012; Verhoeckx and others 2013).
Studies on food safety of cultivated European insects are very limited. Vijver and others (2003) reported upon the uptake and accumulation of heavy metals such as cadmium, copper, lead, and zinc from soils by larvae of T. molitor (Vijver and others 2003). Zagrobelny and others (2009) described that species of Zygaena contained low quantities of cyanogenic glucosides (Zagrobelny and others 2009). Goumperis (2012) presented potential hazards of insects for food and feed application: the presence of pathogens, contaminants (such as pesticides, natural toxins, heavy metals, neoformed substances due to processing, and veterinary residues), allergenicity, and the introduction of pests, and animal and plant diseases into the EU (Goumperis 2012). Recently, Belluco and others (2013) published a review on food safety of European and non-European insects, demonstrating the safety of some insects with no additional hazards in comparison with usually consumed animal products.
Food safety of non-European insects has been studied more widely (Table 1). Reported food safety hazards of non-European insects include allergens (Baldo and Panzani 1988; Blum 1994; Phillips and Burkholder 1995; Vetter 1995; Freye 1996; MacEvilly 2000; Arlian 2002; Lian and Liu 2006; Ji and others 2008; Ji and others 2009), ANFs (Wirtz 1984; Nishimune and others 2000; Adeduntan 2005; El Hassan and others 2008), mycotoxins (Simpanya and others 2000; Braide and others 2011), pesticides (Saeed and others 1993; DeFoliart 1999), heavy metals and alkali (Green and others 2001; Handley and others 2007; Zhuang and others 2009; Banjo and others 2010), natural toxins (Duffey 1980; Wirtz 1984; DeFoliart 1992; Berenbaum 1993; Blum 1994; Nishimune and others 2000; Zagrobelny and others 2009), and the presence of pathogenic microorganisms (Simpanya and others 2000; Amadi and others 2005; Giaccone 2005; Banjo and others 2006; Templeton and others 2006; Braide and others 2011; Klunder and others 2012). Examples of bacteria found in insects are species from the genera Staphylococcus and Bacillus (Amadi and others 2005; Banjo and others 2006; Braide and others 2011), Campylobacter (Templeton and others 2006), Pseudomonas (Banjo and others 2006), Micrococcus and Acinetobacter (Amadi and others 2005), Proteus and Escherichia (Braide and others 2011), Enterobacteriaceae, and certain sporeforming bacteria (Klunder and others 2012). Examples of fungi found in insects are Aspergillus, Penicillium, Fusarium (Simpanya and others 2000; Braide and others 2011), Chaetomium, Mucor, Mucorales, Alternaria, Drechslera, and Phoma (Simpanya and others 2000). Insects may also contain pathogens as a result of improper processing or handling (Banjo and others 2006).
Table 1. Food safety hazards of non-European insects
Country of origin
T. molitor (mealworm) and Zophobas morio (superworm)
Toxic chemicals are acquired in 2 ways, either by autonomous production of defense chemicals (such as toxins and toxic metabolites) or by sequestering phytochemicals directly from the food plant (Duffey 1980; Wirtz 1984; Berenbaum 1993; Blum 1994; Schabel 2008). Defensive secretions that may be reactive, irritating, or toxic include, among many others, carboxylic acids, alcohols, aldehydes, alkaloids, ketones, esters, lactones, phenols, 1,4-quinones, hydrocarbons, and steroids. Phytochemicals sequestered by various insects include phenolics, flavin, tannins, terpenoids, polyacetylenes, alkaloids, cyanogens, glucosinolates, and mimetic amino acids (Wirtz 1984). Some insects may contain chemicals in concentrations higher than acceptable levels for food consumption (Yen 2008). For example, arsenic was accumulated in Bogong moth from agricultural sprays, such as the herbicide monosodium methylarsenate (Green and others 2001), and selenium was accumulated in T. molitor (Hogan and Razniak 1991). Food processing can also introduce toxic substances by chemical reactions of substrates of insects and other ingredients, such as heterocyclic aromatic amines, acrylamide, chloropropanols, and furans (Dolan and others 2010).
Based on these studies on food safety of non-European insects, food safety issues should be identified and assessed before insects can enter the market in Europe. Both chemical and microbiological hazards can be introduced or formed in concentrations that may be harmful for public health. Feed for insects (feed, vegetables, and waste products) can be contaminated with mycotoxins, natural toxins, heavy metals, veterinary residues (including antibiotics), pesticides, and pathogens. Potential hazards of the insects themselves can be allergens, pesticides, contaminants, and pathogens. During rearing, insects may be able to convert or accumulate contaminants present in their feed, which can result in degradation or increase of the concentration of substances. The particular safety hazards depend on the insect species, their feed and environment, and production methods. Therefore, more attention should be directed toward the effects of these environmental and management conditions on the safety of insects destined for human or animal consumption.
Algae belong to a large and diverse group of organisms using photosynthesis, which do not belong to the group of terrestrial plants (Cazaux and others 2010). Algae can be distinguished as microalgae and seaweed (Kerkvliet 2001; Stegeman and others 2010). Microalgae are single-celled organisms that can grow over a wide range of environmental conditions, whereas seaweeds are complex multicellular organisms growing in salt water or a marine environment (Cazaux and others 2010).
About 2% of 4000 varieties of algae can form neurotoxins and hepatotoxins that can accumulate in shellfish, crustaceans, and fish. This may result in diseases such as paralytic shellfish poisoning (PSP) due to saxitoxin, diarrhetic shelfish poisoning (DSP), neurotoxic shellfish poisoning (NSP) due to brevetoxins, ciguatera fish poisoning (CFP) due to ciguatoxin/maitotoxin, amnesic shellfish poisoning (ASP), and microcystin (Roheim and others 1995; Kerkvliet 2001; Vershinin and Orlova 2008; Caron and others 2010; Gerssen and others 2010; Van der Fels-Klerx and others 2012a, b).
About 30% of the current worldwide algal production is sold for animal feeding purposes (Becker 2004). Algae are approved in several countries as chicken feed and do not require new testing or approval for feed use (Becker 1994). With regard to food use, the marine diatom Odontella aurita by Innovalg (France) has been approved since 9 December 2002 as a novel food (Gouveia and others 2008).
Several microalgae (for example, species from the genera Chlorella, Tetraselmis, Spirulina, Nannochloropsis, Nitzchia, Navicula, Chaetoceros, Scenedesmus, Haematococcus, and Crypthecodinium) can be used in feed for both terrestrial and aquatic animals (Harel and Clayton 2004).
Microalgae used for human consumption are Arthrospira (spirulina, namely, cyanobacteria), Chlorella spp., Dunaliella salina, and Aphanizomenon flos-aqua (Kerkvliet 2001; Spolaore and others 2006). They originate from several countries: Spirulina from, for instance, China, India, Japan, and United States (US) (Spolaore and others 2006; Small 2011), chlorella from, for instance, Taiwan, Germany, and Japan, Dunaliella salina from Australia, Israel, US, China, and Aphanizomenon flos-aqua from the US (Spolaore and others 2006). The green algae (Chlorophycea) Chlorella vulgaris, Haematococcus pluvialis, Dunaliella salina, and the cyanobacteria Spirulina maxima are widely commercialized and used, mainly as nutritional supplements for humans and as animal feed additives (Kerkvliet 2001; Gouveia and others 2008; Stegeman and others 2010).
Several studies show that safety hazards related to algae may include allergens, toxins, pathogens, heavy metals, and pesticides.
Allergenicity has been reported for airborne cyanobacteria Phormidium fragile and Nostoc muscorum (Sharma and Rai 2008), and the green algal genus Chlorella (Tiberg and Einarsson 1989). However, a high-lipid product, Whole Algalin Flour, composed of dried milled Chlorella protothecoides showed little potential for food allergy (Szabo and others 2012).
No toxins have been found in spirulina and chlorella. However, in Aphanizomenon flos-aqua, toxic microcystines have been detected (Kerkvliet 2001; Heussner and others 2012). Extracts from products consisting of Aphanizomenon flos-aquae, Spirulina, and Chlorella, or mixtures thereof, were cytotoxic (Heussner and others 2012). Under favorable conditions, pheophorbides are formed in chlorella, which give rise to photosensibilization in some humans (Kerkvliet 2001).
Another safety aspect is the presence of pathogenic microorganisms. Spirulina and chlorella are cultivated in open bassins, which may result in microbiological contamination from birds, insects, and rodents (Kerkvliet 2001).
Algae may accumulate heavy metals. Since algae are at the bottom of the aquatic food chain pyramid, they are the most important vector for transfer of pollution to upper levels of the trophic chain in aquatic environments (Souza and others 2012). Sludge-grown algae contain a rather substantial amount of heavy metals that may impose adverse effects to higher trophic organisms (Hung and others 1996; Wong and others 1996). Spirulina accumulates more heavy metals than chlorella (Kerkvliet 2001). Other authors found no exceedance of legal maximum levels of heavy metals as established in their own jurisprudence (Hsu and others 2001; Kolb and others 2004; Marles and others 2011).
The pesticide fenamiphos and its metabolites can be transformed and accumulated by Pseudokirchneriella subcapitata and Chlorococcum spp. Therefore, contamination of natural environments can have adverse impacts on the food chain and associated biota (Caceres and others 2008).
Seaweed species that are used for direct consumption include, among others: Ulva rigida, Monostroma sp., Enteromorpha sp., Laminaria sp., Undaria pinnatifida, Hizikia fusiforme, Himanthalia elongata, Eisenia bicyclis, Ascophyllum nodosum, Fucus vesiculosis, Porphyra sp., Cladophora glomerata, Microspora floccosa, and Palmaria palmata (Kerkvliet 2001; Fahprathanchai and others 2006; Murphy 2007; Akköz and others 2011). Seaweeds can be harvested from the sea, but they are also increasingly cultivated (Cazaux and others 2010). Seaweed products in fresh and dried form are imported from Japan and France. Laminaria sp. (kelp weed) originating from the Arctic Ocean, Iceland, and Norway are mainly used in food supplements (Kerkvliet 2001). No research has been performed on the extraction of proteins of seaweeds for human consumption (Stegeman and others 2010). Several seaweed species, such as U. rigida, Ascophyllum, Laminaria, Undaria, Porphyra, Cystoseira barbata, and C. glomerata have been evaluated as feed additives in meal to fish diets (Appler and Jauncey 1983; Fahprathanchai and others 2006; Kut Güroy and others 2007).
The use of C. glomerata, M. floccosa, and P. palmata in daily meals has been safe for humans (Fahprathanchai and others 2006; Mouritsen and others 2013). Several studies show that safety hazards for seaweed may include iodine, ANFs, heavy metals, radioactive isotopes, ammonium, dioxins, and pesticides.
In Laminaria, high levels of iodine were found (Van Netten and others 2000; Kerkvliet 2001). A broad diversity of seaweed species (red, green, and brown) contains ANFs, such as low levels of lectins, tannins, and phytic acid, and high levels of trypsin inhibitors and amylase inhibitors (de Oliveira and others 2009).
Seaweeds can accumulate heavy metals depending on the growing time in the sea and the levels of heavy metals in the water (Ortega-Calvo and others 1993; Murphy 2007; Smith and others 2010). The types and concentration of metals found in seaweed vary with species, collection time, growth phase, and collection site (Houa and Yanb 1998; Brown and others 1999; Van Netten and others 2000; Murphy 2007; Smith and others 2010). Reported accumulation of heavy metals includes arsenic (Van Netten and others 2000; Kerkvliet 2001; Almela and others 2002; Rose and others 2007; Besada and others 2009; ), copper (Prasher and others 2004; Murphy 2007; Murphy and others 2007, 2009; Riosmena-Rodriguez and others 2010; Akköz and others 2011), cadmium (Prasher and others 2004; Besada and others 2009; de Oliveira and others 2009; Akköz and others 2011), chromium (Murphy 2007; Murphy and others 2008, 2009; de Oliveira and others 2009), nickel (Prasher and others 2004; de Oliveira and others 2009; Akköz and others 2011), vanadium (de Oliveira and others 2009), iron, magnesium (Riosmena-Rodriguez and others 2010), mercury (Van Netten and others 2000), lead ( Van Netten and others 2000; Prasher and others 2004; Akköz and others 2011), cesium-137, and radium-226 (Van Netten and others 2000). The uptake of heavy metals for P. palmata was found to decrease in the order: lead > cadmium > copper > nickel (Prasher and others 2004). Iron, copper, and magnesium were the most significant metals found in red and green algae (Riosmena-Rodriguez and others 2010). In general, brown algae have higher arsenic levels than red or green algae (Almela and others 2002). Other studies found no harmful quantities of heavy metals (Hwang and others 2010; Smith and others 2010; Dhaneesh and others 2012). Little is known about the process of metal bioaccumulation in marine food chains because data on metal concentrations at different trophic levels and their temporal or spatial variation are sparse (Van Netten and others 2000; Riosmena-Rodriguez and others 2010; Akköz and others 2011).
Seaweeds also accumulate ammonium due to a combination of the size of the plant and the spatially and temporally variable concentration of ammonium in the seawater (Rees 2003).
Other contaminants found in seaweeds are dioxins and pesticides. Seaweeds were contaminated by polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) of industrial origin, such as combustion source (Hashimoto and Morita 1995). Pesticides that could be present in seaweeds are, for example, azametiphos, diflubenzuron, teflubenzuron, propoxur (Lorenzo and others 2012), and organic micropollutants (polychlorinated biphenyls (PCBs), chlorinated pesticides, polycyclic aromatic hydrocarbons (PAHs)) (Pavoni and others 2003). Further studies regarding the presence of chemotherapeutic agents in seaweeds are required, because of the lack of data on some currently used compounds, and to elucidate the transformation and biodegradation processes (Lorenzo and others 2012).
Duckweed species are small floating aquatic plants. They are monocotyledons of the botanical family Lemnaceae and are higher plants or macrophytes (Leng and others 1995). Duckweed family comprises of 4 genera (Lemna, Spirodela, Wolffia, and Wolfiella) and 34 species (Rahman and Hasegawa 2011). Duckweed species are found worldwide; they are adapted to a wide variety of geographic areas and climatic zones. They are found in all areas except waterless deserts and those permanently frozen. They grow best in tropical and temperate zones, but many species can survive extreme temperatures (Leng and others 1995).
Duckweeds have been supplemented to feed, especially to complement diets and to increase animal growth. Duckweed is used as feed for fish (for example, carp and tilapia) (Fasakin and others 1999; Leng 1999; Yilmaz and others 2004) and domestic animals, including poultry (Haustein and others 1992; Leng 1999; Ahammad and others 2003), ducks (Ikaheimonen and others 1997; Leng 1999; Ngamsaeng and others 2004), and pigs (Hang 1998; Leng 1999; Aguilera-Morales and others 2005). Few studies have been performed on duckweed meal as supplement to forages given to ruminants. It appears that duckweeds can be used as a mineral (particularly P) and N source. The combination of crop residues and fresh duckweeds in a diet for ruminants could give a balance of nutrients. These diets can be potentially applied in cattle, sheep, and goat production systems (Leng and others 1995).
Besides a component of animal and bird diets, duckweed is already a human food resource in traditional/small farmer systems in South Asia (Leng 1999; Adeduntan 2005; Derksen and Zwart 2010); Wolffia arrhiza has traditionally been eaten as “khai-nam,” in Burma, Laos, and northern Thailand. Khai-nam is generally regarded as a poor people's food and so far has attracted little attention as a potentially significant source of human food (Bhanthumnavin and McGarry 1971).
Safety hazards for duckweed may include the presence of heavy metals, phenols, pesticides, dioxins, and pathogens.
The accumulation of heavy metals by duckweed poses a potential danger where heavy metal contamination of water occurs (Leng 1999; Chandra and Kulshreshtha 2004). Heavy metals can enter the food chain at a number of points, and therefore, these contaminants have to be monitored during the production of duckweed for food/feed purposes (Leng 1999; Hoving and others 2011). For instance, crayfish fed with cadmium-containing duckweed were contaminated with cadmium (Arrhenius and others 2006). Accumulation of heavy metals from water includes cadmium (Gaur and others 1994; Boonyapookana and others 2002; Arrhenius and others 2006; Seth and others 2007; Razinger and others 2008; Derksen and Zwart 2010), selenium (Zayed and others 1998), copper (Jain and others 1989; Gaur and others 1994; Chandra and Kulshreshtha 2004; Miretzky and others 2004; Razinger and others 2007; Kanoun-Boulé and others 2009; Derksen and Zwart 2010), chromium (Staves and Knaus 1985; Gaur and others 1994; Boonyapookana and others 2002; Chandra and Kulshreshtha 2004; Miretzky and others 2004; Derksen and Zwart 2010), nickel (Gaur and others 1994; Axtell and others 2003; Derksen and Zwart 2010), lead (Gaur and others 1994; Gallardo-Williams and others 2002; Axtell and others 2003; Miretzky and others 2004; Derksen and Zwart 2010; Sobrino and others 2010), zinc (Gaur and others 1994; Miretzky and others 2004; Derksen and Zwart 2010; Hoving and others 2011), cobalt (Gaur and others 1994), iron (Jain and others 1989; Chandra and Kulshreshtha 2004; Miretzky and others 2004; Derksen and Zwart 2010), arsenic (Seth and others 2007; Rahman and others 2008; Hoving and others 2011; Rahman and Hasegawa 2011; Hoving and others 2012), uranium (Hogan and others 2010), manganese (Miretzky and others 2004; Derksen and Zwart 2010), aluminum, gold, and strontium (Derksen and Zwart 2010). Duckweed has been demonstrated to be a good accumulator of cadmium, selenium, and copper, a moderate accumulator of chromium, and a poor accumulator of nickel and lead (Zayed and others 1998). The uptake of metals depends on the chemical form present and on the life form of the macrophytes (floating, free floating, well rooted, or rootless) (Chandra and Kulshreshtha 2004).
Duckweed species are also capable to uptake and transform phenols (Fujisawa and others 2010) and pesticides, including organophosphorus pesticides (malathion, demeton-S-methyl, and crufomate) (Gao and others 2000), lipophilic compounds (De Carvalho and others 2007), 3-methyl-4-nitrophenol, 3,5-dichloroaniline, 3-phenoxybenzoic acid (Fujisawa and others 2006), dimethomorph (Olette and others 2008; Dosnon-Olette and others 2010), copper sulphate and flazasulfuron (Olette and others 2008), and xenobiotics (chlorophenols) (Day and Saunders 2004).
Limited work has been done to characterize secondary metabolites in specific species of duckweed. So far, oxalic acid is the only identified compound produced by duckweed that is toxic to animals at high levels (Adeduntan 2005).
Holshof and others (2009) found dioxins in harvested duckweed, probably due to the presence of animal proteins of diving beetles or snakes (Holshof and others 2009).
Pathogens like Escherichia coli or Clostridium botulinum could contaminate duckweed (Hoving and others 2011). Islam and others (2004) studied the fecal coliform contamination of duckweed grown on hospital-based wastewater. They found that wastewater-treated duckweed may be safely used as fish feed (Islam and others 2004). Moyo and others (2003) concluded that the use of duckweed for chicken feed was microbiologically safe with respect to E. coli and Salmonella spp. provided that caution is taken during the processing of duckweed. Chickens may get contaminated, especially during wet weather due to poor environmental sanitation at the plant (Moyo and others 2003). Duckweed production in open greenhouse pools would be expected to produce biomass with an associated microflora of bacteria, viruses, fungi, algae, and possibly, microscopic invertebrates (Adeduntan 2005).
For using duckweed protein products for food or animal feed, methods would need to be developed to insure that heavy metals, phenols, pesticides, and pathogens would not contaminate the protein products (Stomp 2005; Hoving and others 2011).
Rapeseed (Brassica napus) is a bright yellow flowering member of the Brassicaceae. In 1986, the definition of canola was introduced to refer to B. napus and B. campestris (now Brassica rapa) lines containing less than 2% erucic acid in the oil and less than 30 μmol/g glucosinolates in the air-dried, oil-free meal (OECD 2011).
Rapeseed or canola proteins have been used as feed ingredient around the world for many years for a broad range of animal species including poultry, pig, cattle, and fish (including salmon, trout, tilapia, and prawns) (Burel and others 2000; Enami 2011; OECD 2011; Nagel and others 2012). For human food, canola proteins have been characterized to have interesting functional properties that may replace classical ingredients in different food formulations (Aluko and McIntosh 2005; Yoshie-Stark and others 2006; Cumby and others 2008; Guo and others 2010; Aider and Barbana 2011). However, the application of canola protein as a human food ingredient has been limited to only a few food products manufactured and marketed on a low scale in Canada, Japan, and the US (such as processed meats, cheeses, pizza, and bagels) due to technological limitations in relation to organoleptic properties and ANFs (Mejia and others 2009).
In the US, generally recognized as safe (GRAS) notifications on canola protein ingredients have been received (ADM 2010; Keller-and-Heckman 2011). Mejia and others (2009a, b) found, in specific canola/rapeseed protein isolates, that potential natural toxicants such as glucosinolates and erucic acid, as well as potential contaminants such as pesticide residues, solvent residues, heavy metals, dioxins, aflatoxins, PAHs, and acrylamides, were either nondetected or below toxicological and regulatory allowed limits (Mejia and others 2009). However, in the EU, an application for a protein preparation isolated from B. napus and B. rapa was not accepted as a novel food due to the lack of limit values for relevant undesirable compounds, identification of the starting material, and studies on allergic reactions (CBG-MEB 2012).
Several studies showed that safety hazards for rapeseed may include ANFs, heavy metals, and allergens. Rapeseeds contain several ANFs, such as erucic acid, glucosinolates, phytic acid, phenolics (mainly sinapine and tannins), and a high fiber content (Kozlowska and others 1990; Burel and others 2000; Bonnardeaux 2007; Mejia and others 2009; Aider and Barbana 2011; OECD 2011). One of the limiting factors for the application of canola proteins is that the content of phenolic acids in canola meals is up to 5 times higher than in soybean meals, and in rapeseed/canola flours 10 to 30 times higher than in flours from other oleaginous seeds such as flaxseed (Aider and Barbana 2011). Pig and poultry feed-producing industries are not receptive to using canola meal for full protein supplementation because of the ANFs in canola meal (Bonnardeaux 2007). However, modern technologies used in processing (such as chemical modifications, microbial and physical treatments, membrane filtration, as well as a development of low-glucosinolate and low-erucic acid rapeseed cultivars) are able to eliminate the majority of the ANFs in canola such as glucosinolates, phytates, and tannates (Mansour and others 1993; Aider and Barbana 2011).
Heavy metals from the soil can accumulate in the roots, plant, and seeds of rapeseeds. Based on pot experiments, lead and zinc from the soil can accumulate in the roots of Brassica napus L., and small amounts of them move through the conductive system to the seeds. Cadmium moves relatively easily from root to stem and is accumulated in higher concentrations in the top of the plant (Angelova and others 2008). Rossi and others (2004) also showed that B. napus accumulated zinc and copper and translocated these elements in different ways in the harvestable parts of the plants (Rossi and others 2004). Ahmad and others (2011) found that lead, cadmium, and chromium concentrations in soil, forage, and seed of Brassica napus L. increased after treatment with sewage water (Ahmad and others 2011). Brunetti and others (2011) found that B. napus was able to accumulate high amount of metals in greenhouse conditions, in the order: chromium > zinc > copper > lead.
Rapeseed contains allergenic 2S storage proteins (napins) (Monsalve and others 2001; Poikonen and others 2006). These proteins show a cross-reactivity with related Brassica species (such as mustard) (Monsalve and others 1997, 2001; Focke and others 1998), since these proteins exhibit great sequence similarity with 2S albumins from different seeds. The 2S albumin seed storage proteins in mustard seeds share 94% sequence similarity with 2S albumins from rapeseed (Monsalve and others 2001). Several allergic reactions in humans have been reported after consumption of mustard and rapeseed products (Meding 1985; Widström and Johansson 1986; Monsalve and others 2001; Poikonen and others 2006, 2008, 2009; Hafting and others 2012). Currently, mustard is included in the list of 14 allergenic foods that must be declared on food labels of prepackaged foods in the EU according to Directive 2006/142/EC.
For use of rapeseed protein products for food production, methods would need to be developed to insure that heavy metals would not contaminate the protein products. The effects of ANFs and allergens should also be taken into account.
Overview of potential hazards in novel protein sources
Table 2 shows an overview of the potential hazards in the 5 novel protein sources considered. Heavy metals and processing contaminants are potential hazards for all 5 protein sources. Pesticides, pathogens, and allergens can be present in most protein sources. In addition, insects and seaweed have also other potential hazards like ANFs, mycotoxins, and dioxins.
Table 2. Overview of potential hazards in 5 novel protein sources (insects, microalgae, seaweed, duckweed, and rapeseed), as based on literature sources
Food and feed business operators (FBOs) that wish to put novel food and feed protein products on the market have to comply with European and national rules for food and feed. For the novel products discussed in the previous sections below, some comments will be made on the general legal requirements as regard to market introduction and food and feed safety legislation.
All FBOs in the EU should comply with the requirements established in EU and national law. Requirements are set for the producer (including transporters), the food and feed products, and the presentation of these products. Most of the requirements apply to all FBOs and food and feed products (horizontal law), some only to specific products (vertical law).
The General Food Law (Regulation (EC) 178/2002,1 GFL) sets the framework for EU legislation on food and feed, and applies to all stages of food and feed production. First of all, this Regulation gives the definition of what is considered to be food and feed. Both definitions are very broad. Food or foodstuff is defined as “any substance or product, whether processed, partially processed or unprocessed, intended to be, or reasonably expected to be ingested by humans” (art. 1). Feed or feedingstuff is defined as “any substance or product, including additives, whether processed, partially processed or unprocessed, intended to be used for oral feeding to animals” (art. 2). According to the GFL, the FBO is responsible for compliance of products, processes, and premises with all requirements of food law (art. 17). Foods (and feeds) should not be placed on the market if they are unsafe (art. 14 and 15) and must be traceable by the FBO (art. 18). Furthermore, in case a FBO has reasons to believe that a product is unsafe, it should immediately withdraw the product from the market and inform the competent authorities (art. 19 and 20). The presentation of the product should not mislead consumers (art. 16). The GFL as such defines the general rules for FBOs.
Food and feed hygiene legislation
The Food Hygiene Regulations, Regulation (EC) 852/2003 and (EC) 853/2004 contain general rules on hygiene of foodstuffs for all manufactures of food, including those involved in primary production. Only production for private domestic consumption or handling, and the direct supply of small quantities of primary products to consumers or local retail establishments are excluded from the scope of this Regulation. According to the Hygiene regulations, the FBO is obliged to notify the competent authorities of its existence (art. 6, 852/2004). When FBOs are handling certain products of animal origin, they should be approved before start of operations (art. 4, 853/2004). Also, FBO involved in feed production should be either registered or approved (art. 9 and 10, Feed Hygiene Regulation (EC) 183/2005) by the competent authorities of a member state. For registration, a FBO should inform the competent authority (CA) on the establishment and/or activities. For an approval, the CA must visit and give an approval prior to the upstart of the business.
Requirements for FBOs in primary production are laid down in Annex I, and for FBOs in other stages of production in Annex II of Regulation (EC) 852/2004. FBOs in primary production are required to take measures to control contamination of their products (Annex I, Part A. Regulation (EC) 852/2003), such as the requirement to use potable or clean water whenever necessary to prevent contamination, to use biocides, plant protection products, veterinary medicinal products, and feed additives correctly, and to keep records of measures taken to control hazards. Those FBOs producing primary products of animal origin should also keep records on, among others, the nature and the origin of feed fed to animals (Annex I Part A., III 8). FBOs involved in the production, processing, and distribution of food after the stage of primary production should have food safety programs and procedures based on HACCP principles (art. 5 Regulation (EC) 852/2004). National and Community guides on good hygiene practices for control of hazards, including those in primary production, are (being) developed to help FBOs in this respect. Member states should assess these guidances to confirm that they assure compliance with hygiene regulations. National guides on good hygiene practice are compiled in a register of the EU Commission (Belluco and others 2013). For nonprimary producers and transporters, Annex II of Regulation (EC) 852/2004 contains general requirements for premises (like adequate number of lavatories), equipment and personnel, cleaning and maintenance, rooms where foodstuffs are handled, for transport, food waste, foodstuffs, and water supply. For FBOs involved in the production of food of animal origin, more elaborate hygiene requirements are laid down in Regulation (EC) 853/2004. However, the most extensive requirements in this regulation are only applicable to those animals that are specifically mentioned in Annex I. For a number of pathogenic microorganisms in specified foodstuffs, microbiological criteria are laid down in Regulation (EC) 2073/2005, for novel food products not included in the categories in this regulation, no microbiological criteria have been established yet.
For FBOs producing feed, Regulation (EC) 183/2005 applies. Annex I of this regulation contains, among others, hygiene provisions and the obligation of record keeping for those involved in the primary production of feed. For FBOs involved in feed after the stage of primary production, requirements on facilities and equipment, personnel, production, quality control, storage and transport, record-keeping and complaints, and product recall are laid down in Annex II. An obligation for the latter FBOs is that they shall monitor for the presence of prohibited and undesirable substances and other contaminants. Furthermore, feed FBOs should, like food FBOs, have procedures in place based on hazard analysis and critical control point (HACCP) principles. For assistance in the application of HACCP principles, community and national guides to good practice have been developed and made available (Aider and Barbana 2011).
Novel food legislation
Novel food and novel food ingredients that were not consumed “to a significant degree” in the EU prior to 15 May 1997 have to be authorized before market introduction according to Regulation (EC) 258/97, the Novel Food Regulation (NFR). At the time of drafting of the NFR, no EU definition of “food” existed in the EU (Jones 2012); therefore, 4 categories of novel food and food ingredients to which the NFR applies were identified in art. 1 (2) c–f (a and b concerning genetically modified organisms were later moved to separate regulations) (see our italics):
c.Foods and food ingredients with a new or intentionally modified primary molecular structure.
d.Foods and food ingredients consisting of or isolated from microorganisms, fungi, or algae.
e.Foods and food ingredients consisting of or isolated from plants and food ingredients isolated from animals, except for foods and food ingredients obtained by traditional propagating or breeding practices and having a history of safe food use.
f.Foods and food ingredients to which has been applied a production process not currently used, where the process gives rise to significant changes in the composition or structure of the foods or food ingredients which affect their nutritional value, metabolism, or level of undesirable substances.
Most novel protein products mentioned in this publication will fall within category d or e. Although it is not always clear to what category a new food product belongs in reality, this does not make a huge difference in the data that have to be provided by the applicant for the safety assessment (Jones 2012).
The producer of a novel food has to notify a member state of the EU its intention to place a novel product on its market for the first time. The CA of a member state then checks if the food is a novel food and if the dossier is complete, and if so, makes a first assessment of the product. This assessment is forwarded to the European Commission (EC) which forwards the assessment to the CAs in all member states. If no objections are made by member states, an authorization is granted; if reasonable objections are made, then the dossier is forwarded to European Food Safety Authority (EFSA). Based on the EFSA opinion, a decision on authorization is taken under the EU committee procedure involving, among others, the Standing Committee on the Food Chain and Animal Health (SCFCAH). An authorization is granted to the applicant of the novel food, and not to all FBO wishing to put a similar new product on the market of the EU. For “substantially equivalent” novel products, however, the applicant can and should notify the EC. In practice, for a substantially equivalent product, a statement of the CA of a member state confirming the equivalence should accompany this notification. The outline of and explanations on both procedures are given in a recently published decision tree on the internet (WageningenUR 2013).
The assessment aims to establish whether the novel food, or novel food ingredient, is safe for the consumer, does not mislead the consumer, or when it replaces another ingredient, is not nutritionally disadvantageous for the consumer (art. 3.1). For the assessment, the applicant has to provide all the necessary data; data requirements are further specified in Recommendation 97/618/EC.
Since the entering into force of Regulation (EC) 258/97, experience has revealed that the procedures and requirements are not fully transparent for applicants of novel foods and novel food ingredients. Main uncertainties concern definitions and data requirements. The regulation applies to placing on the market any novel food and food ingredient, in which novel is defined as “which have not hitherto been used for human consumption to a significant degree within the Community…” “Not hitherto” means before May 15, 1997. Both the date and the borders of the EU were arbitrarily chosen, resulting in some products not being “novel,” while other similar products had to be assessed for safety (Verhagen and others 2009). CAs from member states interpreted “the Community” as all member states of the EU, irrespective of their date of entry. This interpretation was confirmed in the recently published Information and Guidance document (see below). The expression “used for consumption to a significant degree” is also not clear, since nowhere in the regulation is explained what is meant by “to a significant degree.” The SCFCAH decided that presence in food supplements prior to May 15, 1997, would not be considered to constitute consumption to a significant degree (SCFCAH 2005). An example of use of this guidance is the ruling on chromium picolinate by the SCFCAH (SCFCAH 2011). Just recently, the Directorate General for Health & Consumers of the European Commission (DG SANCO) published an “Information and Guidance Document on Human Consumption to a Significant Degree” (Naidu and others 1999). In this document, a decision tree is presented and a questionnaire, to be filled in by the applicant, on consumption aspects to help applicants to identify and justify significant consumption. The United Kingdom (UK) and Belgium also provide some advice on how to prove that a food is not novel on its Internet site (ANCFP 2005). Examples of proof of nonnovelty are invoices, legal import documents, lists of prices of food products dated before May 15, 1997, literature data, dated labels, FAO statistics, certificates or declarations of other member states on the status of a food, and so on. It is pointed out that the provided evidence should clearly demonstrate oral consumption of the food product by humans in the EU. The “Novel Food Catalogue” (EC 2013), a searchable database that lists products that have already been assessed before, as to being novel or not, could also help FBOs to determine the status of a product.
Besides administrative information, extensive data on composition, nutritional value and metabolism, intended use, and intake of and levels of hazardous substances in the novel food are essential. Furthermore, microbiological and toxicological (including allergenicity) information on the novel food should be provided by the applicant. For new products like insect proteins, the data requirements on composition, like analyses of a number of representative batches regarding nutrients, micronutrients, and known and potential contaminants, might be problematic if only pilot studies have been performed with breeding and harvesting of insects. Especially for proteins, the allergenic potential should be considered. For new proteins expressed in genetically modified crops, an EFSA opinion has been published for the assessment allergenicity (EFSA 2010a). This opinion could be used as a starting point in the assessment of non-GM new proteins. Opinions of EFSA on novel protein products like “alfalfa protein concentrate” (EFSA 2009), “sardine peptide product” (EFSA 2010b), and bovine lactoferrin (EFSA 2012) further illustrate data requirements for novel proteins.
(Novel) Feed legislation
Feed materials are defined as “products of vegetable or animal origin, whose principal purpose is to meet animals” nutritional needs, in their state, fresh or preserved, and products derived from industrial processing thereof, and organic and inorganic substances, whether or not containing feed additives, which are intended for use in oral animal-feeding either directly as such, or after processing, or in the preparation of compound feed, or as carrier of premixtures’ (art. 3 (2g) Regulation (EC) 767/2009). The term “feed material” is further explained in a Recommendation (2011/25/EU) that was published with guidelines to clarify the distinction between feed materials, feed additives, and other feed products. In a quality control system for feed producers, a decision tree was developed to guide producers in definitions used in feed legislation (GMPplus 2011). Annex III of Regulation (EC) 767/2009 lists materials prohibited for feed use.
There is no premarket authorization procedure for novel feed products. A FBO just has to notify appropriate representatives of the European feed business sectors that the intention is to place a novel feed product on the EU market for the first time, that is, from 1 September 2010 onwards (art. 26(1) Regulation (EC) 767/2009), and for products that were on the market before September 1, 2010, but not listed in the catalogue (see further). This notifying FBO has to be registered as a FBO (art. 11, Regulation (EC) 183/2005). The representatives should publish the notifications on the Internet and update the register of feed materials with each notified novel feed material (art. 24(6) Regulation (EC) 767/2009). This Feed Materials Register (Feedmaterialsregister 2010) was set up solely for information purposes. The safety of the notified novel feed material is not assessed, neither by the representatives nor competent authorities. Thus, in the first instance, it is the FBO that is responsible for the safety of the new product. In the register, feed names are not unique, so the same name may be used for feed materials with different characteristics. However, products mentioned in the Feed Catalogue (Regulation (EU) 68/2013) are only allowed to be marketed with the name in the catalogue, provided they are compliant with the descriptions of the product and labeling requirements to the catalogue. The representatives can make amendments of the catalogue, to include products from the register, on a voluntary basis. Once taken up in the catalogue, the products with the same name in the register will be removed. The fact that a product is mentioned in the register or catalogue does not imply that it can be used in the feed of all animals. Animal proteins, for instance, are, with some exceptions, not allowed to be used in the feed of food-producing animals as a result of the “extended feed ban” (see further under insects).
Legislation on chemical food and feed safety
For certain hazardous chemicals, like pesticides and veterinary drugs, an application for authorization must be filed with the competent authorities prior to use. If necessary, legal maximum limits will be set for residues of these chemicals in foods. Other chemicals occur naturally, are formed during processing, or are present in food and feed due to their presence in the environment. For some of these chemicals, maximum limits are also set by law, and there are limits for “undesirable substances” in feed. This legislation is listed in Table 3.
Table 3. EU legislation on hazardous substances in food and feed products
Food / feed products
Regulation (EC) 396/2005
Residues of plant protection products
Primary agricultural products that are used as food and feed and are listed in Annex I
Regulation (EU) 37/2010
Residues of veterinary medicinal products
Muscle meat, liver, kidney, fat (or skin and fat), milk, egg, and honey used as food
For plant protection products, the maximum residue limits (MRLs) set apply to the products listed in Annex I of Regulation (EC) 396/2005 (Regulation (EU) 212/2013), whether they are used as food or as feed. In general, the products listed in Annex I are primary agricultural products. It is foreseen that for some processed products, “specific concentration or dilution factors” will be laid down in Annex VI of Regulation (EC) 396/2005, but no factors have been established yet. For products not listed in Annex I, no EU MRLs for pesticide residues exist. This is also the case for the product groups “fish, fish products, shell fish, molluscs and other marine, and fresh water food products” and “crops or parts of crops exclusively used for animal feed” in Annex I. For the latter groups, MRLs are not applicable until individual products in these groups have been listed, but no individual products have been listed up till now. Seaweeds and “other terrestrial animal products” are included in Annex I. Thus, also for food and feed derived from insects, pesticide MRLs apply, although in most cases, no specific MRLs will have been set and the default MRL of 10 ppb or limits of quantification will apply.
According to the veterinary drug residues legislation (Regulation (EC) 470/2009), food-producing animals are animals that are bred, raised, kept, slaughtered, or harvested for the purpose of producing food for humans. The only product of an insect that is included in the list of animal products is honey from bees. In Volume 8, the guidance note for applicants of MRLs for veterinary drugs, the other animal products mentioned are: liver, kidney, fat (fat and skin for pigs), and (injection site) muscle from mammals; liver, kidney, fat and skin, and muscle of poultry; and muscle and skin from fish (CVMP 2005). The animal products mentioned are the only products of animals for which MRLs are set thus far in the MRL regulation, Regulation (EU) 37/2010.
For contaminants, legal maximum levels (MLs) are only set for those food products that are contributing to human exposure. Levels are set as low as reasonably achievable (Regulation (EEC) 315/93). New food products (not on the market yet) do not contribute yet to human exposure. Thus, the regulation on contaminants in food, Regulation (EC) 1881/2006, does not yet contain MLs for the 5 novel protein foods considered. For oils and fats derived from plants and animals, also novel fats and oils, there are MLs for lead, and the sum of 3 PAHs, and benzo(a)pyrene. For oils of marine organisms and vegetable oils and animal fats, there are MLs for dioxins, dioxin-like PCBs, and PCBs. Also, for “food supplements” that may contain new food products, MLs have been set for a limited number of contaminants.
For new products used as or in feed, Directive 2002/32/EC does contain maximum levels for a rather high, compared to food, number of undesirable substances. This is because feed products in this directive are not specified in as much detail as in the food contaminants regulation. Most new products will fall in the broad category “feed materials” of this directive. This means that for use as feed, there are maximum levels for arsenic, cadmium, fluorine, lead, mercury, nitrite, melamine, aflatoxin B1, several inherent plant toxins, the persistent pesticides aldrin, dieldrin, chlordane, DDT, endosulfan, endrin, heptachlor, hexachlorobenzene, hexachlorocyclohexane (alpha, beta, and gamma isomers), dioxins and dioxin-like PCBs, and PCBs for all new products that are used as feed material. Furthermore, harmful botanical impurities (among others, several mustards) that should not be present in feed are listed in this directive.
In the next section, more specific remarks will be made on legislation applying to novel proteins derived from animal sources (insects) and plant sources.
Specific legislation for novel protein sources
When farmed insects or products derived from farmed insects are produced to be used as food for humans, or as feed for food-producing animals, a FBO has to comply with rather complex legal requirements. These requirements are related to the insects themselves, the feed or substrate fed to the insects, the firm producing the insects, and the ultimate marketing for use as food or feed.
In the Netherlands, it is forbidden to keep animals for agricultural production that are not listed in the Dutch Decision on the indication of animals that are allowed to be kept for production purposes (Overheid 1997). Several insects, including those proposed to be used in food or feed, are listed in this decision. The regulation on protection of animals at the time of killing, Regulation (EC) 1099/2009, does not mention insects. Reptiles and amphibians are excluded from this regulation, because these “are not animals commonly farmed in the Community,” so maybe insects will be excluded also in future.
It is not clear if food products consisting of or derived from insects are considered to be “products of animal origin” as defined in the Hygiene Regulations (EC) 852/2004 and (EC) 853/2004. No specific requirements are laid down for the production and processing of insects in these regulations. As no requirements for insects are laid down in Annex III of Regulation (EC) 853/2004, establishments handling insects do not need approval prior to the start of operation (but they should be registered). The breeding of insects is part of “primary production,” the processing of insects into food is not. Thus, FBOs breeding and converting insects into food should comply with Annex I and II of Regulation (EC) 852/2004.
Whether feed, sometimes called substrate, fed to insects that are bred for consumption by humans or food-producing and nonfood-producing animals, should comply with Regulation (EC) 767/2009 is not clear. A “food-producing animal” in this regulation is defined as “any animal that is fed, bred, or kept for the production of food for human consumption, including animals that are not used for human consumption, but that belong to a species that is normally used for human consumption in the Community” (art. 3 (2c)). Insects are “not normally” used for human consumption in the EU. Annex III of this regulation lists a number of materials that are prohibited for use as feed, such as feces and “household waste,” materials that could be used to feed insects. On the opposite, feed or substrates for insects probably do have to comply with EU regulations on animal proteins, Regulation (EC) 1069/2009 (and the implementing Regulation (EU) 142/2011) as the definition of “farmed animals” in this regulation does not exclude insects: “any animal that is kept, fattened, or bred by humans and used for the production of food, wool, fur, feathers, hides, and skins or any other product obtained from animals or for other farming purposes.” These regulations prohibit the use of some (animal protein) sources that might be suitable as feed for insects, like those mentioned under Category 2 materials: manure and gut content, dead-in-shell poultry, and fallen stock (art. 9). Also, the use of “catering waste” as feed for farmed animals (apart from fur animals) is prohibited (art. 11b). Of special concern when feeding insects are genetically modified (GM) plants with built-in insecticides, like the crops in which insecticidal proteins of the microorganism Bacillus thuringiensis (Bt) are expressed. These GM crops may be detrimental to the growth of insects. All GM crops should be labeled according to EU legislation, however; thus, in principle, use of Bt-derived ingredients could be avoided.
Although insects are already sold as food in the EU, mostly in specialty shops, the discussion as to whether insects and insect-derived products are novel foods in the EU is still ongoing. Furthermore, it is not clear if the NFR applies to whole insects used as food, since in the definition only “food ingredients isolated from animals” are considered to be novel (Belluco and others 2013). The Food Standards Agency (FSA) in the UK launched a survey in August 2011 on the consumption of insects in the UK, to determine if consumption “in a significant degree” before May 15, 1997, could be substantiated with data. This would obviate the need for a market application under the NFR for those insects for which a significant degree of consumption could be proven. The results of this survey are not published yet.
The legislation on the use of insects and insect-derived products as feed for food-producing animals is complicated. In the past, the use of animal proteins in feed for food-producing animals was banned for safety reasons, for example, the prevention of transmission of TSE/BSE. The initial ban on use of mammalian proteins in feed for ruminants was later extended to the use of all animal proteins, also from nonmammalian sources, in feeds of all food-producing animals (with some minor exceptions). The question is thus whether insect-derived proteins are animal proteins. The fact that insect meal and insects are included in category 9.16.1. in the Catalogue of Feed materials does not imply that these products are therefore allowed to be used in feed for food-producing animals. According to an answer given March 25, 2010, by Commissioner Dalli on a question from the European Parliament, insect meal is a “processed animal protein” as referred to in Annex IV of the Regulation (EC) 999/2001 and is therefore prohibited to be fed to farmed animals. Annex IV of this regulation contains details concerning the prohibitions (part I) and derogations (part II) on the feeding of animal proteins to categories of food-producing animals, and as such give details for the prohibitions in art. 7.1. (ruminants) and art. 7.2 (other animals). There is neither explicit prohibition of, nor derogation for, the use of proteins from terrestrial invertebrates in Annex IV. Contrary to the answer by Dalli in a subsequent communication of the EU Commission, “TSE road map 2” (EC 2010), it is indicated that animal proteins not specifically excluded are allowed to be used in feeds of all nonruminants. In the table in this Roadmap, insect proteins are not specifically excluded.
Regulation (EC) 142/2011 (implementing Regulation (EC) 1069/2009) gives a definition for “processed animal protein” (PAP). In this definition, PAP is a product derived from “Category 3 material” treated in a certain way. Some specific products are excluded from the definition, such as milk, eggs, and products derived thereof. In article 10 of Regulation (EC) 1069/2011, a description of Category 3 material is given. This category includes, among others, all by-products that are the result of the production of food products derived from animals fit for human consumption, but also products from terrestrial invertebrates (not pathogenic to humans and animals, art. 10(l)). Thus, it is here that insect-derived proteins are included in PAP. Proper processing of insects to convert them to “processed animal proteins” is a basic requirement for use of these proteins in feed. Possible processing strategies are listed in Annex IV of Regulation (EC) 142/2011. If just grinding and drying of whole insects or insect larvae, part of primary production, will be considered as proper processing by competent authorities is not clear, although the CA has sometimes some room to interpret.
Nowadays, new analytical control tools have been developed and a relaxation of the ban, allowing proteins from nonruminants to be used in feed again, was recently published. Regulation (EU) 56/2013 allows the use of nonruminant proteins in feed for fish in aquaculture. As transmission of BSE from nonruminants, also nonruminants like insects, to other nonruminants is negligible (Regulation (EU) 56/2013, recital 6), a lifting of the ban on use of insect proteins in the feed of food-producing pigs and poultry should be on the agenda of the Commission in the near future.
Foods consisting of or derived from insects fall under the group “other terrestrial animal products” of the pesticide residue legislation (Annex I, Regulation (EC) 396/2005). This means that either specific MRLs will have been set, or the default MRL or limits of quantification are applicable. It might be necessary to treat insects raised for food use for diseases like infestation with other insects, fungi, bacteria, or viruses. Pesticides that are used to treat animals, and thus insects fall under the veterinary drugs legislation. A pharmacologically active substance of a veterinary drug must be listed in Table 1 of the Annex of Regulation (EU) 37/2010 and, if necessary, MRL must have been set in food products before a veterinary drug can be registered in a member state. No products derived from insects, apart from honey, are present in this regulation, so there are no MRLs for such products. If substances contained in Table 1 of the Annex are found in the official control in matrices for which no MRLs have been set, member states are expected to make a scientific assessment of the risk, and forward this assessment to the Commission. The registration procedure for the use of veterinary drugs on/in insects will probably be complicated and of a long duration, as no data requirements have been formulated yet. For insects raised solely for feed, the veterinary residue regulations do not apply. But, of course, products of animals fed with insects or insect-derived products should comply with the veterinary drug residue legislation. Insect products used as feed should also comply with MLs mentioned in Directive 2002/32/EC.
Foods consisting of or derived from algae are included in the definition of novel foods in the NFR. Thus, prior to market introduction of novel algae, an application should be filed for authorization of the product. Some products of algae might have been used as food prior to May 15, 1997. The Novel Food Catalogue should provide some insight as to whether an alga is novel or not. Several substances derived from algae are used as food additive. For food additives, Regulation (EC) 1333/2008 applies. In the past, prior to market access, a FBO was obliged to apply for authorization for the use of proteins derived from algae in feed under Directive 82/471/EEC. Assessment of safety and nutritional value was done according to the guidelines in Directive 83/228/EEC. To assist applicants, EFSA published a “Guidance for the assessment of biomasses for use in animal nutrition” in 2011. Regulation (EC) 767/2009 repealed both directives, and to date, no formal authorization and safety assessment is required anymore. But the Guidance of EFSA might still be useful for FBOs to assess their products.
Algae are likely to belong to the product group “fish, fish products, shell fish, molluscs, and other marine and fresh water food products” in Annex I of Regulation (EC) 396/2005. MRLs for residues of plant protection products will be set as soon as individual products of this group are defined, which is not the case yet. Algae or their products used as feed should comply with all the MLs mentioned in Directive 2002/32/EC.
Seaweeds are macroalgae, and thus the NFR will apply if the seaweed is novel. Whether a certain variety of seaweeds is considered to be novel or not can be verified to a certain extent in the Novel Food Catalogue. For instance, the brown seaweed Fucus vesiculosus is not a novel food according to this catalogue, but the red seaweed Rhodymenia palmata L (P. palmata) is, as this seaweed has only been used before in food supplements. For proteins isolated from these algae, however, a novel food application might be necessary (art. 1(2f)). For use in feed no authorization is needed. Seaweeds are mentioned in Annex I of Regulation (EC) 396/2005. MRLs for residues of plant protection products only apply if seaweeds are used for human consumption, not if they are used as feed for animals. For parts of seaweeds or seaweeds used exclusively for animal feed, separate MRLs will be set in the future. Only for food supplements consisting exclusively, or mainly, of dried seaweed, and products derived from seaweed, a ML for cadmium has been set in Regulation (EC) 1881/2006. Fresh seaweed is explicitly excluded in this regulation as a category of food for which the MLs of lead and cadmium are applicable. For seaweed meal and feed materials derived from seaweed, a separate ML for arsenic is set in Directive 2002/32/EC. All other products of seaweed used as feed should comply with the other MLs mentioned in Directive 2002/32/EC.
Common duckweed, Lemna minor, was considered to be a novel food when an application was filed under the NFR (Novel Food Catalogue). Also, other subspecies of duckweed are likely to be novel foods and should be authorized prior to marketing as a food. For use in feed, no authorization is required. Duckweed is likely to belong to the product group “fish, fish products, shell fish, molluscs, and other marine and fresh water food products” in Annex I of pesticide residue Regulation (EC) 396/2005. Like for algae, no MRLs for residues of plant production products have been set, as no individual products have been defined yet for this group. Duckweed products used as feed should comply with all the MLs mentioned in Directive 2002/32/EC.
Oil from rapeseed, or canola, has been used as food for humans for some time now and is thus not a novel food. For this oil use, special rapeseed varieties are grown with low levels of the toxic fatty acid erucic acid (Directive 80/891/EEC) and glucosinolates. This is different for proteins derived from rapeseed seeds. Although leftovers from rapeseed processing have been used in animal feed, no use of products other than oil is common in food for humans. For rapeseed proteins, the NFR applies. The oilseeds of rapeseed are included in Annex I of Regulation (EC) 396/2005. Thus, MRLs for plant protection product residues have been laid down in the other annexes of this regulation. For parts of rapeseed other than the seeds, the MRLs are not applicable. For oil from rapeseed, MLs have been set in Regulation (EC) 1881/2006 for the contaminants lead, benzo(a)pyrene, dioxins and dioxin-like PCBs, and PCBs. For the feed material called rapeseed cakes, a specific ML has been set in 2002/32 for allyliosthiocyanate (an indicator of the presence of volatile mustard oil). All other products of rapeseed should comply with the other MLs mentioned in Directive 2002/32/EC.
Unclarities in EU legislation
Table 4 shows an overview of the unclarities in EU legislation for the novel protein sources. Most unclarities in EU legislation were found for insects as novel protein source. For algae, seeweed, and duckweed, some MRLs are not yet set. No unclarities were found for rapeseed.
Table 4. Overview of unclarities in EU legislation for the novel protein sources insects, algae, and duckweed
Unclarity in EU legislation
Are insects and insect derived products novel foods?
MRLs for residues of plant protection products are not yet set
Regulation (EC) 396/2005; Directive 2002/32/EC
For (parts of) seaweeds used exclusively for animal feed, separate MRLs will be set in future
Regulation (EC) 396/2005; Directive 2002/32/EC
Control of Safety Hazards
Producers of novel proteins are responsible for the application of a novel food dossier or for risk assessment of their new products. They are obliged to prove that their products are safe. Although some safety aspects of novel proteins are intrinsic to the product, many potential hazards can be controlled by production methods and by production and processing conditions. The selection of a species or a variety of the protein source can affect safety due to differences in allergenicity, metabolism, and composition. This has already been applied to rapeseed, currently containing less erucic acid and glucosinolates than before. As feed of insects may contain several contaminants that insects may accumulate and metabolize, the selection of feed is important to prevent contaminants in the human and animal diet. For example, the composition of waste streams from industry is variable and can therefore introduce contaminants in the production chain. The same applies for microalgae, seaweed, duckweed, and rapeseed, all are able to accumulate heavy metals from the environment. These plants have to be produced using controlled conditions. Good Manufacturing Practices (GMP)+ requirements can be used to develop a controlled production system. Processing conditions of novel protein products (as an ingredient or as a product as such) can also affect the safety of novel proteins. Heating of products containing novel proteins can result in the formation of new components (neoformed components), like acrylamide. Also, the application of solvents for the extraction of proteins can affect safety of the novel protein products. During product development, these aspects should be considered in advance.
Novel protein sources, like insects and algae, are expected to be increasingly used in Europe as replacers for animal-derived proteins. Technical and processing properties are being investigated but, to date, possible food safety hazards associated with the use of novel proteins in feed and food applications are hardly known. These hazards may include a range of contaminants, like heavy metals, mycotoxins, pesticide residues, as well as pathogens. Food business operators that wish to put on the European market products derived from novel proteins should comply with European legislation and possibly additional national legislation. To date, European law is not conclusive on several issues regarding the use of novel protein sources in feed and food products. One of the major unclarities for feed applications is whether or not products with insects are considered animal-derived products or not. If so, it seems to be possible to put on the markets feed material using novel protein sources. For food product applications, the most important question for food producers is whether or not the product is considered a novel food. If so, the producer has to provide a Novel Food Dossier, among others, proving the product is safe for the consumer. In all cases, the producer is responsible for the safety of the product, but due to the unclarities in European law, it is not always clear which Regulation and maximum levels for contaminants apply. In order to stimulate the use of novel protein sources in Europe, it is therefore advised to adjust and clarify the European legislation. Future research should focus on effects of raw materials and the environment on the safety of novel proteins in food and feed products, the degradation and accumulation of several substances during processing of these products, and the transmission of substances to feed and food products.
The authors acknowledge the Ministry of Economic Affairs (EZ) for financial support of this study (KB-15-007-003).
M. van der Spiegel drafted the part of novel protein sources as well as their potential hazards, M.Y. Noordam described the legislation in general and specific for novel protein sources, and H.J. van der Fels-Klerx stated the conclusions and critically reviewed the manuscript.
The legal acts quoted in this section refer, where applicable, to the latest amended version.