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The potential of various insect species for use as food for fish
The potential of various insect species for use as food for fish Fernando G. Barroso, Carolina de Haro, Mar´ıa-Jos´e S´anchez-Muros, Elena Venegas, Anabel Mart´ınez-S´anchez, Celeste P´erez-Ba˜no´ n PII: DOI: Reference:
S0044-8486(13)00679-0 doi: 10.1016/j.aquaculture.2013.12.024 AQUA 630970
To appear in:
Aquaculture
Received date: Revised date: Accepted date:
18 June 2013 11 December 2013 12 December 2013
Please cite this article as: Barroso, Fernando G., de Haro, Carolina, S´ anchez-Muros, Mar´ıa-Jos´e, Venegas, Elena, Mart´ınez-S´anchez, Anabel, P´erez-Ba˜ n´on, Celeste, The potential of various insect species for use as food for fish, Aquaculture (2013), doi: 10.1016/j.aquaculture.2013.12.024
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The potential of various insect species for use as food for fish
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Fernando G. Barroso 1*, Carolina de Haro 1, María-José Sánchez-Muros 1, Elena Venegas 2, Anabel Martínez-Sánchez3 & Celeste Pérez-Bañón3
Department of Applied Biology, University of Almería, Almería, Spain.
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Food Technology Division, University of Almería, Almería, Spain.
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Instituto Universitario CIBIO, University of Alicante, Alicante, Spain.
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* Corresponding author:
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Fernando G. Barroso
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Department of Applied Biology,
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E.S.I. University of Almería,
Carretera de Sacramento s/n, 04120 Almería, Spain
Fax: +34950015476
Phone: +34950015918
e-mail: [email protected]
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ACCEPTED MANUSCRIPT Abstract Due to the expansion of aquaculture and the limited resources available from the sea, it
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is necessary to find substitutes for fish meal for use in aquaculture. We believe that the
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use of insect meals as an alternative source of animal protein may be an option. To use insects for this purpose, it is necessary to determine the nutritive characteristics of these
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insects. To determinate the potential of insects as a substitute for fish meal in fish food
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used in aquaculture, we examined 16 different species, 5 of then as different stage of development, of the orders Coleoptera (4), Diptera (7) and Orthoptera (5). The insect
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analysed have a higher proportion of fat and less protein than fish meal. With the exceptions of histidine, threonine and lysine, the insects present an amino acids profile
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similar to fish meal, being Diptera b the most similar group to fish meal. However, the
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fatty acid content of insects is very different from that of fish meal that is rich in n-3,
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specially 14% EPA, 16% DHA, which are practically absent in insects, how have higher ratios of omega 6 and monounsaturated fat.
KEY WORDS: fish nutrition, alternative feed, insects meal, fish meal, amino acids, fatty acid.
1. Introduction Fish have been a key source of food for humans (Ayoola, 2010), but the global catch of wild fish declined approximately 3% from 2004 to 2009 at a rate of 0.5% per year (FAO, 2010). Currently, aquaculture is playing an essential role in the seafood market, meeting the demand for fish that cannot be met with the wild catch. As a consequence,
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ACCEPTED MANUSCRIPT in recent years (from 2004 to 2009), aquaculture production has grown by 32%, a growth rate of approximately 5.6% per year (FAO, 2010).
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Fishmeal is one of the major components of the feed used in aquaculture. It is
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generally added to animal diets to increase feed efficiency and animal growth through better feed palatability; it also enhances the uptake, digestion, and absorption of
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nutrients (Mile and Chapman, 2006). It is estimated that approximately 30% of the total
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fish catch is converted to fish meal and fish oil for use in animal and fish feeds (Ogunji et al., 2006).
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The percentage of fish meal that is used for aquaculture feeds has increased from 10% in 1988 to approximately 45% in 2002. The increasing global demand for and
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decreasing availability of fish meal has led to sharp increases in the price of fish meal,
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and hence, the cost of aquaculture production has increased as well (Ayoola, 2010). The
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price of producing fish through aquaculture has risen from US $600/metric ton in 2005 to US $2000/metric ton in June 2010, and this trend is likely to continue (International Monetary Fund, 2010).
The present shortage of fish meal motivates researchers to seek new protein sources with nutritional values similar to fish meal, in particular those with similar contents of the essential amino acids, phospholipids, and fatty acids (docosahexaenoic acid and eicosapentaenoic acid) that promote optimum development, growth, and reproduction (Ayoola, 2010), which would allow aquaculture production to remain economically and environmentally sustainable over the long term. From vegetable sources, soybean meal is the best available vegetable protein source in terms of protein content and EAA profile. However, it is potentially limiting in sulphur-
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ACCEPTED MANUSCRIPT containing amino acids (methionine and cysteine) and contains some antinutrient substances such as trypsin inhibitor, haemagglutinin, and antivitamins (Tacon, 1993).
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Regarding sources of animal origin mostly of them are forbidden by prescription of
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food security, which have made more urgent the search for alternatives to fish meal in aquaculture diets (Ogunji, 2004).
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Edible insects are a natural renewable resource used as food by humans (Ramos-
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Elorduy and Conconi, 1994). Since ancient times, insects have been one alternative protein source used to compensate for the periodic or seasonal scarcity of other sources
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(Ramos‐Elorduy, 1997). The most existing studies have focused on the insects that have played an important role in human nutrition in Africa, Asia, and Latin America. Thus,
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we can highlight the studies conducted in Nigeria (Akinnawo and Ketiku, 2000; Banjo
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et al., 2006), Mexico (Ramos-Elorduy and Conconi, 1994; Ramos-Elorduy et al., 1997;
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Ramos-Elorduy et al., 2006; Ramos‐Elorduy, 1997), Thailand (Yhoung‐Aree et al., 1997) and Zaire (Kitsa, 1989). Although insects began to be evaluated as a potential foodstuff for animals 40 years ago (Calvert et al., 1969; Hale, 1973; Ichhponani and Malik, 1971; Newton et al., 1977; Phelps et al., 1975; Teotia and Miller, 1974), the incorporation of insects into fish feed has not received much attention until recently (Ogunji et al., 2006). In the last 10 years there have been several studies of feeding experiments performed in vivo with diets based on insect meal in Clarias anguillaris (Achionye-Nzeh and Ngwudo, 2003), Clarias gariepinus (Alegbeleye et al., 2012; Aniebo et al., 2011; Fasakin et al., 2003), Orcorhychus mykiss (Sealey et al., 2011; St-Hilaire et al., 2007; ) Oreochromis niloticus (de Haro et al., 2011a; de Haro et al., 2011b; de Haro et al., 2011c; de Haro et al., 2011d; de Haro et al., 2011e; Ogunji et al., 2006; Ogunji et al., 2008) and Psetta
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ACCEPTED MANUSCRIPT maxima (Kroeckel et al., 2012). In general, percentages of substitution higher than 30% decreased the growth depending on the fish and insect species.
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From the nutritional point of view, depending on species and/or stage, insects are rich in
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protein and lipids, nevertheless the presence of chitin a priori indicates a negative characteristic. However, chitin also is present in crustacean, which are widely consumed
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by fish.
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To evaluate the potential of insects, it is necessary to consider other advantages such as environmental benefits; the insects can be fed with waste generated by humans, having
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an important role in recycling materials in the terrestrial biosphere (Katayama et al., 2008). The great diversity of insect species (70-75% of animal species), from different
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ecosystems, with different diets and stages of development (larval, pupa, ninpha or
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imago) provokes a huge variability in body composition.
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The aims of this study are: (1) to determine the nutritive valour of the most frequent rearing insects and some of the common species of Almeria (Spain), and (2) to establish the most similar to fish meal.
2. Materials and methods 2.1. Sampling
The insects and stages of its development (larval, pupa, ninpha or imago) studied were chosen according to the following criteria: easy to rearing, stage with mayor biomass and low exoskeleton. The wild species were chosen in terms of their availability in the environment. The insects used in the study were obtained from pet shops (captivity*), reared by the research group "Bionomía, Sistemática e Investigación Aplicada de insectos dípteros e
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ACCEPTED MANUSCRIPT himenópteros" of the University of Alicante (captivity **) or captured in the field close
belong to three orders, Diptera, Orthoptera and Coleoptera.
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to our environment (wild-rearing). Table 1 summarises the insects studied, which
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The nutritional contents of insects were compared with those of fish meal and soybean
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meal because they are the most common ingredients used in aquafeed production.
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2.2. Analytical methods 2.2.1. Determination of proximate composition
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The nutritional values obtained were derived from three replicate samples for each species of insecta, fish meal and soy meal. Moisture, crude protein, total lipids and ash
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were determined using AOAC (2005) techniques.
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The insects were sacrificed by freezing (Finke et al., 1989). All samples, insects,
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fishmeal and soy meal were lyophilised (Cryodos, Ima-Telstar, Terrassa, Spain), and ground and freezing until to be analized. Total nitrogen (N) was determined using the Kjeldahl procedure, and crude protein was estimated as N x 6.25. Crude lipid was determined following the Soxhlet extraction of dried samples with petroleum ether. Moisture was determined after oven drying the samples at 105 ºC to a constant weight. The ash was determined eliminating the organic matter at 500ºC during 12h.
2.2.2. Determination of amino acid profile The amino acid profile was determined after hydrolysing the sample with 6 N HCl for 22 h at 110 ºC, followed by a sequence of filtering, derivatisation, and separation in a gas chromatograph. Tryptophan was not determined.
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ACCEPTED MANUSCRIPT 2.2.3. Analysis of fatty acids For the FA analyses, all samples were transmethylated following the method of Lepage
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and Roy (1984) with the minor modifications of Venegas-Venegas et al. (2011): for
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each sample, 1 ml of freshly prepared transesterification reagents (methanol/acetyl chloride, 20:1, v/v) was added to 50 mg of freeze-dried insect meal in a glass tube along
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with 100 µl of a solution of internal standard (heptadecanoic acid 17:0, 10 mg/ml). The
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tubes were shaken and then placed in a hot block (100 ºC, 30 min). Next, the mixture was cooled to room temperature, and 1 ml of distilled water was added to each tube.
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The samples were shaken again and centrifuged (3,000 rpm, 3 min). The upper hexane phase was collected for GLC analysis.
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The resulting FA methyl esters (FAMEs) were analysed in a Focus GLC (Thermo
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Electron, Cambridge, UK) equipped with a flame injection detector (FID) and an
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Omegawax 250 capillary column (30 m 9 0.25 mm i. d. 9 0.25 lm film thickness; Supelco, Bellefonte, PA, USA). The temperature program was 1 min at 90 ºC, heating to 200 ºC at a rate of 10 ºC/min, constant temperature at 200 ºC (3 min), heating to 260 ºC at a rate of 6 C/min and constant temperature at 260 ºC (5 min). The injector temperature was 250 ºC with a split ratio of 50:1. The injection volume was 4 µl. The detector temperature was 260 ºC. Nitrogen was used as the carrier gas (1 ml/min). Total saturated, monounsaturated, polyunsaturated, n-3 and n-6 fatty acids were calculated as the sums of saturated fatty acids (ΣSFA), monounsaturated fatty acids (ΣMUFA), polyunsaturated fatty acids (ΣPUFA), n-3 and n-6 fatty acids, respectively.
2.3. Data analysis
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ACCEPTED MANUSCRIPT To better understand the applied value of the study’s results, in addition to a descriptive approach to the nutritional value of the insects, the compositions of the insect meals
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were compared with the compositions of fish meal and soybean meal.
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To determine the similarity between the compositions of fish meal, soybean meal and the different species of insects, a hierarchical cluster analysis was used.
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Clustering is a multivariate technique of grouping together rows that share similar
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values. Cluster analysis is an exploratory data analysis technique for solving classification problems. Its purpose is to sort the different food sources (fish meal,
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soybean meal and insect meals) into groups (clusters) so that the degree of association/similarity between members of the same cluster is stronger than the degree
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of association/similarity between members of different clusters. Each cluster is
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described as the class to which its members belong. This analysis provides a
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dendrogram grouping the meals according to the distance between them. The similarities were evaluated using hierarchical cluster analysis in the Past (Paleontological Statistics, Version 2.17). The analysis was performed using hierarchical option clusters based on Ward’s method (Ward, 1963) with standardised data. Bootstrapping has previously been used to obtain confidence intervals. Specifically, there were two cluster analyses based on different characteristics: •
The proportion of essential amino acids.
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The proportion of fatty acids
We observed that the cluster analysis classified different insect species according to their taxonomic order. Therefore, to analyse the differences between orders, we grouped the data by order regardless of the species. An analysis of variance with the taxonomic order as main factor was performed with the JMP (Version 9.0.0) statistical package
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ACCEPTED MANUSCRIPT (SAS Institute, Inc.) to evaluate the relationship between the compositions of amino acids and fatty acids. Significant differences in the mean response were determined with
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the Tukey-Kramer HSD test.
3. Results
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The proximate analysis of the insect samples is summarised in table 2. As expected,
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most insect species analysed have a high proportion of protein, between 40 and 60% in similar to soy meal levels (50% CP) and lower than fishmeal (73.0% CP). The lowest
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levels were measured in Rhynchophorus ferrugineus (35%) and Hermetia illucens larvae (36%). The protein content of Orthoptera (73% CP) is similar to fish meal
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(73.0%), particularly Heteracris littoralis (74%) and Acheta domestica (73%).
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The lipid levels in fish meal (8.2%) and soy meal (3.0%) are lower than in insects. The
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lipid contents of most of the insects reached 20%. The highest values were obtained for Locusta migratoria, Musca domestica larvae and Tenebrio mollitor with 30% lipids and Zophoba morio with 38.0% lipids. These insects contain between 5 and 20% of nitrogen free extract (NFE). Only three of the species had nitrogen free extracts above 35%. NFE involve carbohydrates, sugars, starches, fiber and chitin (primarily a nitrogen-containing polysaccharide). Regarding ash, fish meal contains a higher percentage (18.0%) than insect meals (except for Hermetia pupae, 19.7%) or soybean meal. The amino acid analyses are summarised in table 3. In general, the amino acid patterns were quite different among species. The dendrogram in figure 1 illustrates the similarity of the essential amino acid profiles of the insects, fish meal and soybean meal. As shown in figure 1, two major groups or
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ACCEPTED MANUSCRIPT cluster were formed; (1) fish meal and Diptera, (2) Orthoptera, soybean meal and Coleoptera. Thus, Diptera appears to be the most similar to fish meal in terms of its
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amino acid composition, especially the larvae of Hermetia, Musca and Eristalis,
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whereas Coleoptera is the most different from fish meal in amino acid composition. Soybean meal is closer to Orthoptera in the dendrogram. These results verified the
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relationship between similarity in amino acid composition and insect taxonomy, as each
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order was grouped into a different cluster.
Figure 2 presents the significant differences in the proportion of essential amino acids
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among the three orders of insects, fish meal and soybean meal for all essential amino acids. Relative to fish meal, the insect meals are deficient in the amino acids histidine,
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lysine and threonine, but better in lysine methionine and tyrosine than soy meal. Diptera
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shows a histidine, lysine and threonine proportion similar to fish meal (figure 2 and
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table 3). Diptera also has a similar proportion of methionine and a higher proportion of phenylalanine compared to fish meal and soy meal. However, Diptera exhibits a relative deficit in leucine that does not occur in Orthoptera or Coleoptera. The percentages of tyrosine and valine were higher in all analysed insects than in fish meal. These data reveal that Diptera has similar amino acids profile to fish meal. Ortopthera and Coleoptera, although not so similar to fish meal, have better amino acids profile than soy meal. Table 4 and figure 3 show fatty acids composition of insect fish meal and soy meal. The most notable difference was the higher percentage of n-3 fatty acid in fish meal, up to 37% of the total fatty acids, containing 14% EPA (20:5 n3) and 16% DHA (22:6 n3). In contrast, the insects have a much lower proportion of n-3, while soy mal a rich in n-6. Some species of Orthoptera show a higher proportion (Hetteracris littoralis, 19.8% and
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ACCEPTED MANUSCRIPT Anacridium aegyptium, 17.9%), but the n-3 in those species consists primarily of aclinolenic acid (ALA 18:3 n3). Chrysomya megacephala and Calliphora vicina (Diptera)
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only contain EPA, but it does not exceed 1.5% of total FAs, a low percentage compared
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with the fish meal values.
The saturated fatty acid compositions of the insect meals are similar to fish meal;
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nevertheless, insects have nearly twice monounsaturated fatty acid than fish meal and
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soy meal . The difference in n-6 is much larger; fish meal contains only 2.5% n-6, and soy meal 55,4%, whereas insects have a intermediate proportion, in some species
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reaching over than 30% (e.g., Tenebrio mollitor, Gryllus assimilis and Acheta domestica). Eristalis tenax is the only studied species that had a proportion (1.9%) of n-
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6 similar to that of fish meal.
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The variability of fatty acid profiles among species (figure 3) was greater than the
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variation in essential amino acids, with significant differences in only some fatty acids. The Orthoptera exhibited a higher proportion of n-3, especially for ALA 18:3 n3. No differences between groups for n-6 acids were observed except for the lower proportion of linoleic acid (18:2 n6) in Diptera compared to Orthoptera. The dendrogram (figure 4) illustrates the similarities of the fatty acid compositions among insects, fish meal and soybean meal. The results of the cluster analysis show several groups and the distances are higher than the amino acids. Farthest group is composed of fish meal, Eristalis tenax and Hermetia ilucens larvae. Soybean meal and the other insects (Diptera, Coleoptera and Orthoptera) are mixed in more distanced groups.
4. Discussion
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ACCEPTED MANUSCRIPT The results obtained of nutrients composition of insect show differences regarding fishmeal and among species and/or stage.
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4.1. Proximal composition
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Most of the insects species analysed have a high proportion of protein, similar to the levels of soy meal but lower than fish meal. In general, the order Orthoptera exhibits a
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higher CP, between 60 and 70%, but it must be taken into account that all samples of
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this order are adults with more chitin (and chitin nitrogen) and less fat that enhances CP levels. In Diptera, the levels of CP range from 40 to 50%, with higher values for the
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larvae of Chrysomya megacephala, pupae of Lucilia sericata and pupae of Protophormia terraenova (50-62%). The CP values vary among published works, the
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CP value obtained for larvae of Musca domestica was 46.9%, similar to that reported by
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Ogunji et al. (2008), 47.1%, whereas other authors have obtained more diverse results,
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including 37.5% (Aniebo and Owen, 2010) and 56.8% (Bernard et al., 1997). In pupae of Musca domestica, Bernard et al. (1997) obtained a higher proportion, 58.3%, than that observed in this experiment, 40.1%. The percentage of CP obtained in larvae of Hermetia illucens (36.2%) is similar to those obtained by Sheppard (2002) (37.8%) and Arango et al. (2004) (37%) and slightly lower than the value of 40.6% described by Newton et al. (1977). The CP content obtained in Eristalis tenax, 40.9%, is similar to that observed by Ramos-Elorduy et al. (1998) in Eristalis sp. (40.7%). In the order Coleoptera, the species with the least CP is Rhynchophorus ferrugineus (35%). The larvae of the genus Rhynchophorus are not very protein-rich; Banjo et al. (2006) reported that Rhynchophorus phoenicis was only 28.4% protein, and Cerda et al. (1999) reported 25.8% protein for Rhynchophorus palmarum. In Tenebrio mollitor larvae, the observed CP was 58.4%, and in Zophoba morio, 53.5%. Both the Tenebrio
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ACCEPTED MANUSCRIPT and Zophoba values are slightly higher than those observed by other authors: between 47 and 53% for Tenebrio mollitor (Bernard et al., 1997; Finke, 2002; 2007; Ramos-
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Elorduy et al., 2006) and 46.8% for Zophoba morio (Finke, 2002).
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These differences may be due to the phase of development of the insect, variations in dietary habits between populations, the method of processing or differences in ecotypes
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(Banjo et al., 2006; Fasakin et al., 2003; Teguia and Beynen, 2005), and this makes it
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difficult to draw solid conclusions related to differences between orders. Nevertheless the results of this study indicate that the protein content of insects is
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generally less than that of fish meal and it is similar to soybean meal. There are substantial differences between the protein contents reported by different authors, even
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for the same species of insect. This must be investigated to know the species and stages
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appropriate to be used in fish feeding.
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On the other hand, an additional problem to CP determination in insect is the nitrogen contained within nacetylglucosamine, a subunit of the chitin polymer, digestibly unavailable (Finke et al., 1989). Finke (2002) suggests that the amount of nitrogen contained in chitin is relatively small, and thus, nitrogen content may provide a reasonable estimate of total protein in most invertebrates. The fat content of insects is highly variable and it seems to vary between stages, usually ranging between 15 and 30%. Zophoba morio is the species with the highest proportion of fat (38%) of all of the analysed species; this value is slightly lower than that obtained by Finke (2002), 42%. The larvae of coleopterans generally have a large amount of fat, often exceeding 25%, the high proportion of fat was recorded for Tenebrio mollitor (30%), lower than levels (38 to 43%) reported by other authors (Finke, 2002; 2007; Ramos-Elorduy et al., 2006). A lower proportion was obtained in this study for the
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ACCEPTED MANUSCRIPT larvae of Eristalis tenax (5.8%), lower than that obtained by Ramos-Elorduy et al. (1998) in Eristalis sp. (11.9%). Fat levels in both, larvae (31.3%) and pupae (33.7%) of
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Musca domestica are greater than those observed by other authors: from 13.5 to 25% in
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larvae (Aniebo and Owen, 2010; Bernard et al., 1997; Ogunji et al., 2006, 2008; Sheppard, 2002) and 15.8% in pupae (Bernard et al., 1997). As reported by Arango et
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al. (2004), the percentage obtained for the larvae of Hermetia illucens, 18%, is lower
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than the values observed by Newton et al. (1977) and Sheppard (2002), both greater than 30%.
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The stage of development may also affect the fat content. In general, the larval stages contained significantly more fat than adults (Barker et al., 1998). In Acheta domestica
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varies between 14 and 22% (Bernard et al., 1997; Finke, 2002; 2007) at different stages
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of development. Probably the differences in lipids among the same species, reported by
age.
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different authors, could be related not only to the stage of development but also to the
Diet appears to be another factor that influences the fat content of insects. A comparison of the fat content of the wild orthopteran Heteracris littoralis, at 8.2%, with captivebred orthopterans (Acheta domestica, Gryllus assimilis and Locusta migratoria), with a higher proportion of fat, suggests that diet could affect lipid content. As it occurs with CP, the lipids content varies enormously. Because of this, it is difficult to obtaine a clear conclusion regarding relation between lipids content and taxon, stage or feeding. Supplementary studies are needed. On the other hand, the lipid levels in fish meal and soy meal are lower than in insects, which could complicate the use of insect meals in fishfeeds.
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ACCEPTED MANUSCRIPT The ash contents obtained for all analysed insects were less than that of fish meal (18.0%), with the exception of Hermetia illucens pupae (19.7%). Arango et al. (2004)
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(17.5%) and Newton et al. (1977) also found elevated levels (14.6%) of ash in this
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species.
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4.2. Amino acid profile
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A balanced essential amino acid (EAA) profile is one of the characteristics that define the quality of protein. The development of commercial aquatic feeds has traditionally
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been based on FM as the primary protein source due to its high protein content and balanced essential amino acid profile (Nguyen et al., 2009) with high level of digestible
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essential amino acids such as lysine, methionine and leucine, which are often deficient
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in grains, the typical base for most animal feeds (Hall, 1992; Keller, 1990). Currently,
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the specific amino acid composition of most insect species is unknown. In this work the amino acids profile of different species and stages have been studied. The results show that the profile of amino acids is related to the taxonomic group (figure 1) and Diptera are most similar, in terms of essential and limiting amino acids, to fish meal (Fig. 2). This group has a similar proportion of methionine to fish meal and higher values of histidine, lysine and threonine. Orthoptera and Coleoptera have a higher proportion of leucine. These results are consistent with several studies in which low levels of methionine have been observed in Orthoptera (Bernard et al., 1997; Finke, 2002; 2007) and Coleoptera (Cerda et al., 1999; Finke, 2002; 2007; Ramos-Elorduy et al., 2006) and high levels in Diptera (Newton et al., 1977; Ogunji et al., 2006). Regarding lysine, the proportions obtained for both Coleoptera and Orthoptera, 6%, are similar to those described by
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ACCEPTED MANUSCRIPT Finke (2002, 2007). In Diptera, the lysine level obtained in this work, 8%, coincided with the value previously reported by Newton et al. (1977).
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The proportions of arginine have been shown to be adequate for Diptera and Orthoptera;
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however, the Coleoptera are in deficit. These results differ somewhat from earlier studies, which found that the proportion of arginine in Orthopterans was approximately
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7% (Finke, 2002; 2007) and those in Diptera (Newton et al., 1977; Ogunji et al., 2006)
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and Coleoptera (Finke, 2002; 2007; Ramos-Elorduy et al., 2006) were approximately 5%.
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In terms of similarity to fish meal, the amino acid profile of the Diptera is superior to that of soybean meal, thus Diptera could be a better replacement than soybean meal in
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the formulation of fish feed. Although each species is deficient in some essential amino
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several species of insect.
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acids, it is possible to design a feedstuff that is balanced in amino acids by combining
4.3. Fatty acids
According to Ramos-Elorduy (2008) insects contain higher quantities of polyunsaturated fats (PUFAs) n-6 than fish meal, but lower than soy meal. As in soy, lower levels of polyunsaturated fats n-3 were observed in insect meal analysed regarding fish meal. Bukkens (1997) reported significant quantities of linoleic acid in the fatty acid content of all analysed species of insect. As observed by other researchers (Akinnawo and Ketiku, 2000; Beenakkers and Scheres, 1971; Ekpo and Onigbinde, 2007; Finke, 2002; 2007; Katayama et al., 2008), terrestrial insects do not contain EPA or DHA except for Chrysomya megacephala and Calliphora vicina, which have
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ACCEPTED MANUSCRIPT between 1.3 and 1.5% EPA, far lower proportions than fish meal, which contains up to 14 to 16% EPA.
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The lack of EPA and DHA in insects is one of the most important limitations to its use
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in seawater fish feed, since it involves a limit in their inclusion in aquafeed. Nevetheless there are evidences that the fatty acid profiles of insects most likely reflect
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the fatty acid composition of their food. In wild Hetteracris Littoralis and Anacridium
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aegyptium individuals that consume plants in the field have a high amount of a-linolenic acid (ALA, 18:3 n3), a precursor of the n-3 series, whereas three other species of
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orthopterans, bred in captivity and fed only with flour and bran cereals, present with lower ALA, 18:3 n3. Finke (2002) has achieved FA profiles virtually identical to those
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obtained in this work in Zophoba morio and Tenebrio mollitor, species bred in captivity.
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The fatty acid content of Locusta migratoria determined in this experiment is very
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different from that obtained by Beenakkers and Scheres (1971), who measured 26% oleic acid (18:1 n9) and 21% ALA (18:3 n3), whereas the results of this experiment show 45% oleic acid and 5% ALA. These differences could be related to diet; Locusta used in this experiment are from pet stores, whereas in the work of Beenakkers and Scheres (1971), Locusta were provided with a diet of reed, endive, or pear (supplemented with some reed). In addition, these authors obtained a fair correlation between the percentages on oleic and linoleic acid in the tri- and diglycerides of both diet and body fat. These results indicate that lipid quality could be manipulated by feeding, which amplifies the nutritive values of insects meal. On the other hand in this work we have not sampled any aquatic insect. Freshwater insects have a high proportion of EPA because they ingest freshwater algae, which generally contain ALA. EPA levels above 15% as have been observed by Bell et al. (1994).
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ACCEPTED MANUSCRIPT The data obtained in this experiment agree with Justi et al. ( 2003): fatty acids content of insects is more dependent on diet, in contrast with amino acids, more related to the
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taxon.
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The comparison with fish meal and soy meal reveals lower levels of n-3 and n-6 in
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insect meal.
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Finally, it is interesting to note the following considerations:
- The utilisation of insects has other advantages that cannot be ignored; insects do not
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compete with human resource and can be reared on by-products or human waste as efficient biotransformers to convert abundant low-cost organic wastes into animal
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biomass that is rich in proteins and suitable for use in animal nutrition (Ramos-Elorduy,
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1999; 2005). Regarding soybean production, which is largely oriented toward animal
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feed, insects require fewer natural resources (i.e., land, water, fertiliser) than soybean, the production of which could compete with human food production. - The current aquaculture production system, supported by fish meal, is clearly unsustainable. As reported by the FAO (2010), capture fisheries have been rather stagnant or even declining in some countries, particularly fisheries of wild marine fish stocks; 3% are under-exploited, 12% are moderately exploited, 53% are “fully exploited”, 28% are over-exploited, 3% are depleted and 1% is recovering from depletion. This situation, together with the increase in demand for fish, indicates that fish meal will become a more limiting ingredient in both production and price. Therefore, it is essential to obtain potential alternatives.
5. Conclusion
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ACCEPTED MANUSCRIPT The data obtained in this work indicate an adequate nutritional composition of the insect meals evaluated, for inclusion in fish food. The great variety of insect species, habitats,
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development stages, feeding habits and other characteristics most likely affects insect
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nutritional value and makes insect meal very interesting to study as an alternative to fish meal.
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Although insects generally present some characteristics that do not match with the fish
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meal, the amino acid profile of the Diptera shows that this group of insects could be a possible alternative protein source to be used in aquaculture. Nevertheless, more studies
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are needed to know the digestibility, chitin content and digestive effect, presence of toxic, meal treatments (such as degreasing), adequate mixtures of different insect
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species or to modify the nutritional value of insects by changing their diet or rearing
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condition. Insects can potentially play a fundamental role in animal nutrition, so further
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studies are required on their nutritional value in the context of their use in animal feed.
Acknowledgements
The authors are very grateful to the Consejería de Innovación y Ciencia, Junta de Andalucía (project AGR5273), Fondos Europeos de Desarrollo Regional (FEDER Funds) and Campus de Excelencia Internacional Agroalimentario y del Mar, Ministerio de Educación, by financial support.
References
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ACCEPTED MANUSCRIPT Achionye-Nzeh, C.G., Ngwudo, O.S., 2003. Growth response of Clarias anguillaris
Bioscience Research Communications. 15, 221-223.
PT
fingerlings fed larvae of Musca domestica and soyabean diet in the laboratory.
RI
Akinnawo, O., Ketiku, A.O., 2000. Chemical composition and fatty acid profile of edible larva of Cirina forda (westwood). African Journal of Biomedical
SC
Research. 3, 93-96.
NU
Alegbeleye, W.O., Obasa, S.O., Olude, O.O., Otubu, K., Jimoh, W., 2012. Preliminary evaluation of the nutritive value of the variegated grasshopper (Zonocerus
MA
variegatus L.) for African catfish Clarias gariepinus (Burchell. 1822) fingerlings. Aquaculture Research. 43, 412-420.
D
Aniebo, A.O., Owen, O.J., 2010. Effects of age and method of drying on the proximate
TE
composition of housefly larvae (Musca domestica Linnaeus) meal (HFLM).
AC CE P
Pakistan Journal of Nutrition. 9, 485-487. Aniebo, A.O., Odukwe, C.A., Ebenebe, C.I., Ajuogu, P.K., Owen, O.J., Onu, P.N., 2011. Effect of housefly larvae (Musca domestica) Meal on the carcass and sensory qualities of the mud catfish, (Clarias gariepinus). Advances in Food and Energy Security. 1, 24-28. AOAC, 2005. Official methods of analysis of the Association of Official Analytical Chemists International, 18th edn ed. AOAC International, Gaitherburg, USA. Arango, G.P., Vergara, R.A., Mejía, H., 2004. Análisis composicional, microbiológico y digestibilidad de la proteína de la harina de larvas de Hermetia illuscens (diptera: Stratiomyiidae) en angelópolis-antioquia, Colombia. Revista Facultad Nacional de Agronomía, Medellín (online). 57, Available at: http://redalyc.uaemex.mx/src/inicio/ArtPdfRed.jsp?iCve=179914073009.
20
ACCEPTED MANUSCRIPT Ayoola, A.A., 2010. Replacement of fishmeal with alternative protein Source in aquaculture diets. Thesis Degree of Master of Science. Faculty of North
PT
Carolina State University. North Carolina, USA. Thesis Degree of Master of
RI
Science in Faculty of North Carolina State University.
Banjo, A.D., Lawal, O.A., Songonuga, E.A., 2006. The nutritional value of fourteen
SC
species of edible insects in southwestern Nigeria. African Journal of
NU
Biotechnology. 5, 298-301.
Barker, D., Marianne, P., Fitzpatrick, D., Dierenfeld, E.S., 1998. Nutrient composition
MA
of selected whole invertebrates. Zoo Biology. 17, 123–134. Beenakkers, A.M.T., Scheres, J.M.J.C., 1971. Dietary lipids and lipid composition of
D
the fat-body of locusta migratoria. Insect Biochemistry. 1, I25-I29.
TE
Bell, J.G., Ghioni, C., Sargent, J.R., 1994. Fatty acid compositions of 10 freshwater
AC CE P
invertebrates which are natural food organisms of Atlantic salmon parr (Salmo salar): a comparison with commercial diets. Aquaculture. 128, 301-313. Bernard, J.B., Allen, M.E., Ullrey, D.E., 1997. Feeding captive insectivorous animals: nutritional aspects of insects as food. Nutrition advisory group handbook. Fact sheet 003. Scientific Advisory Group to the American Zoo and Aquarium Association.
Bukkens, S.G.F., 1997. The nutritional value of edible insects. Ecology of Food and Nutrition. 36, 287–319. Calvert, C.C., Martin, R.D., Morgan, N.O., 1969. House fly pupae as food for poultry. Journal of Economic Entomology. 62, 938–939. Cerda, H., Martínez, R., N., B., Pizzoferrato, L., Hermoso, D., Paole, M., 1999. Cría, análisis nutricional y sensorial del picudo del cocotero Rhynchophorus
21
ACCEPTED MANUSCRIPT palmarum (coleoptera: curculionidae), insecto de la dieta tradicional indigena amazónica. Ecotropicos. 12, 25-32.
PT
de Haro, C., Barroso, F.G., Hidalgo, L., Sánchez-Muros, M.J., Venegas, E., 2011a.
RI
Effect in muscle composition of the inclusion of Tenebrio mollitor meal in tilapia feed (Oreochromis nilotica), Aquaculture Europe (Annual meeting of the
SC
European Aquaculture Society): Mediterranean aquaculture 2020, Rhodes,
NU
Greece.
de Haro, C., Barroso, F.G., Hidalgo, L., Sánchez-Muros, M.J., Uclés, F., 2011b.
MA
Nutritive adaptation in Nile tilapia (Oreochromis niloticus) to Tenebrio mollitor meal as protein source diet: effect in fish growth and performance, Aquaculture
D
Europe (Annual meeting of the European Aquaculture Society): Mediterranean
TE
aquaculture 2020, Rhodes, Greece.
AC CE P
de Haro, C., Sánchez-Muros, M.J., Rueda, M., Martínez, R., Barroso, F.G., 2011c. Efectos de la sustitución de la harina de pescado por harina de tenebrio (Tenebrio mollitor) en la alimentación para tilapia del Nilo (Oreochromis niloticus), XIII Congreso Nacional Acuicultura, Barcelona, Spain. de Haro, C., Barroso, F.G., Hidalgo, L., Sánchez-Muros, M.J., Férriz, M., 2011d. Evaluation of Tenebrio mollitor meal as a protein source in tilapia (Oreochromis nilotica) feed: nutritive utilization and effect in fish growth and performance, Aquaculture Europe (Annual meeting of the European Aquaculture Society): Mediterranean aquaculture 2020, Rhodes, Greece. de Haro, C., Barroso, F.G., Hidalgo, L., Rincón-Cervera, M., A., Sánchez-Muros, M.J., 2011e. Effect of adaptation to ingest a diet based in Tenebrio mollitor meal on body composition in Nile tilapia (Oreochromis niloticus), Aquaculture Europe
22
ACCEPTED MANUSCRIPT (Annual meeting of the European Aquaculture Society): Mediterranean aquaculture 2020, Rhodes, Greece.
PT
Ekpo, K.E., Onigbinde, A.O., 2007. Characterization of lipids in winged reproductives
RI
of the Termite Macrotermis bellicosus. Pakistan Journal of Nutrition. 6, 247251.
SC
FAO, 2010. The State of World Fisheries and Aquaculture 2010. FAO, Rome, Italy.
NU
Fasakin, E.A., Balogun, A.M., Ajayi, O.O., 2003. Evaluation of full-fat and defatted maggot meals in the feeding of clariid catfish Clarias gariepinus fingerlings.
MA
Aquaculture Research. 34, 733-738.
Finke, M.D., 2002. Complete nutrient composition of commercially raised invertebrates
D
used as food for insectivores. Zoo Biology. 21, 269-285.
TE
Finke, M.D., 2007. Estimate of chitin in raw whole insects. Zoo Biology. 26, 105-115.
AC CE P
Finke, M.D., DeFoliart, G.R., Benevenga, N.J., 1989. Use of a four-parameter logistic model to evaluate the quality of the protein from three insect species when fed to rats. The Journal of Nutrition. 119, 864-871. Hale, O.M., 1973. Dried Hermetia illucens larvae (Diptera: Stratiomyidae) as a feed additive for poultry. Journal of the Georgia Entomological Society. 8, 16-20. Hall, G.M., 1992. Fish processing technology. in: Ockerman, H.W. (Ed.), Fishery byproducts. VCH publishers, New York, USA, pp. 155-192. Ichhponani, J.S., Malik, N.S., 1971. Evaluation of de‐oiled silkworm pupae meal and corn‐steep fluid as protein sources in chick rations. British Poultry Science. 12, 231-234. International Monetary Fund, 2010. International Monetary Fund Primary Commodity Prices.
23
ACCEPTED MANUSCRIPT Justi, K.C., Hayashi, C., Visentainer, J.V., de Souza, N.E., Matsushita, M., 2003. The influence of feed supply time on the fatty acid profile of Nile tilapia
PT
(Oreochromis niloticus) fed on a diet enriched with n-3 fatty acids. Food
RI
Chemistry. 80, 489-493.
Katayama, N., Ishikawa, Y., Takaoki, M., Yamashita, M., Nakayama, S., Kiguchi, K.,
SC
Kok, R., Wada, H., Mitsuhashi, J., 2008. Entomophagy: A key to space
NU
agriculture. Advances in Space Research. 41, 701-705. Keller, S., 1990. Making profits out of seafood wastes. Proceedings of the International
MA
Conference on Fish By-Products Anchorage. University of Alaska Sea Grant, Alaska, 248 pp.
D
Kitsa, K., 1989. Contribution des insectes comestibles à l'amélioration de la ration
AC CE P
29, 511–519.
TE
alimentaire au Kasaï-Occidental. Zaïre-Afrique: économie, culture, vie sociale.
Kroeckel, S., Harjes, A.G.E., Roth, I., Katz, H., Wuertz, S., Susenbeth, A., Schulz, C., 2012. When a turbot catches a fly: Evaluation of a pre-pupae meal of the Black Soldier Fly (Hermetia illucens) as fish meal substitute — Growth performance and chitin degradation in juvenile turbot (Psetta maxima). Aquaculture. 364– 365, 345-352.
Lepage, G., Roy, C.C., 1984. Improved recovery of fatty acid through direct transesterification without prior extraction or purification. The Journal of Lipid Research. 25, 1391–1396. Mile, R.D., Chapman, F.A., 2006. The benefits of fish meal in aquaculture diets. The fish site.
24
ACCEPTED MANUSCRIPT Newton, G.L., Booram, C.V., Barker, R.W., Hale, O.M., 1977. Dried Hermetia illucens larvae meal as a supplement for swine. Journal of Animal Science. 44, 395-400.
PT
Nguyen, T.N., Davis, D.A., Saoud, I.P., 2009. Evaluation alternative protein sources to
of the World Aquaculture Society. 40, 113-121.
RI
replace fish meal in practical diets for juvenile tilapia, Oreochromis spp. Journal
SC
Ogunji, J.O., 2004. Alternative protein sources in diets for farmed tilapia. CAB
NU
International Publishing (Oxford, UK). Nutrition Abstracts and Reviews. Series B, Livestock Feeds and Feeding. 74, 23N-32N.
MA
Ogunji, J.O., Kloas, W., Wirth, M., Schulz, C., Rennert, B., 2006. Housefly Maggot Meal (Magmeal): An Emerging Substitute of Fishmeal in Tilapia Diets,
D
Conference on International Agricultural Research for Development; Deutscher
TE
Tropentag, Bonn, Germany.
AC CE P
Ogunji, J.O., Kloas, W., Wirth, M., Neumann, N., Pietsch, C., 2008. Effect of housefly maggot meal (magmeal) diets on the performance, concentration of plasma glucose, cortisol and blood characteristics of Oreochromis niloticus fingerlings. Journal of Animal Physiology and Animal Nutrition. 92, 511-518. Phelps, R.J., Struthers, J.K., Mayo, J.L., 1975. Investigations into the nutritive value of Macrotermes falciger (Isoptera: Termitidae). Zoologica Africana. 10, 123-132. Ramos-Elorduy, J., 1999. Insects as intermediate biotransformers to obtain proteins. in: Dickinson-Bannack, F., Garcia-Santaella, E. (Eds.), Homo sapiens: An endangered species, towards a global strategy for survival. Proceedings of the 4th World Academic Conference on Human Ecology 1993, Yucatan, Mexico, pp. 157–165.
25
ACCEPTED MANUSCRIPT Ramos-Elorduy, J., 2005. Insects: a hopeful food source. in: Paoletti, M.G. (Ed.), Ecological implications of minilivestock: potential of insects, rodents, frogs and
PT
snails. Science Publishers, New Hampshire, USA, pp. 263-291.
RI
Ramos-Elorduy, J., 2008. Energy supplied by edible insects from mexico and their nutritional and ecological importance. Ecology of Food and Nutrition. 47, 280-
SC
297.
NU
Ramos-Elorduy, J., Conconi, M., 1994. Edible insects in the world (liste des espèces, lieux de consommation et ethnies qui les consomment), Abstracts 4th
MA
International Congress of Ethnobiology, Lucknow, India, pp. 311. Ramos-Elorduy, J., Pino, J.M., Correa, S.C., 1998. Insectos comestibles del Estado de
D
México y determinación de su valor nutritivo. Anales del Instituto de Biología,
TE
Universidad Nacional Autónoma de México. Serie zoología. 69, 65-104.
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Ramos-Elorduy, J., Moreno, J.M.P., Prado, E.E., Perez, M.A., Otero, J.L., de Guevara, O.L., 1997. Nutritional value of edible insects from the state of Oaxaca, Mexico. Journal of Food Composition and Analysis. 10, 142-157. Ramos-Elorduy, J., Medeiros-Costa, E., Ferreira-Santos, J., Pino-Moreno, J.M., Landero-Torres, I., Ángeles-Campos, S.C., García-Pérez, A., 2006. Estudio comparativo del valor nutritivo de varios coleoptera comestibles de México y Pachymerus nucleorum (Fabricius, 1792) (Bruchidae) de Brasil. Interciencia. 31, 512-516. Ramos‐Elorduy, J., 1997. Insects: A sustainable source of food? Ecology of Food and Nutrition. 36, 247-276. Sealey, W.M., Gaylord, T.G., Barrows, F.T., Tomberlin, J.K., McGuire, M.A., Ross, C., St-Hilaire, S., 2011. Sensory Analysis of Rainbow Trout, Oncorhynchus mykiss,
26
ACCEPTED MANUSCRIPT Fed Enriched Black Soldier Fly Prepupae, Hermetia illucens. Journal of the World Aquaculture Society. 42, 34-45.
PT
Sheppard, C., 2002. Black soldier fly and others for value added manure management.
RI
Athens, GA.: University of Georgia. Department of Entomology and Animal Science.
SC
St-Hilaire, S., Sheppard, C., Tomberlin, J.K., Irving, S., Newton, L., McGuire, M.A.,
NU
Mosley, E.E., Hardy, R.W., Sealey, W., 2007. Fly prepupae as a feedstuff for rainbow trout, Oncorhynchus mykiss. Journal of the World Aquaculture Society.
MA
38, 59-67.
Tacon, A.G.J., 1993. Feed ingredients for warm water fish: Fish meal and other
D
processed feedstuffs. FAO Fisheries Circular No. 856.
TE
Teguia, A., Beynen, A.C., 2005. Alternative feedstuffs for broilers in Cameroon.
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Livestock Research for Rural Development. 17, Art. #34 Online. Teotia, J.S., Miller, B.F., 1974. Nutritive content of house fly pupae and manure residue. British Poultry Science. 15, 177-182. Venegas-Venegas, E., Rincón-Cervera, M.A., Guil-Guerrero, J.L., 2011. Triarachidonoyl glycerol purification process. J Am Oil Chem Soc. 88, 323-328. Ward, J.H., 1963. Hierarchical grouping to optimize an objective function. Journal: Journal of The American Statistical Association. 58, 236-244. Yhoung‐Aree, J., Puwastien, P., Attig, G.A., 1997. Edible insects in Thailand: An unconventional protein source? Ecology of Food and Nutrition. 36, 133-149.
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ACCEPTED MANUSCRIPT Figure legends
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Figura 1. Cluster analysis of amino acids similarity among insect species, fish meal and
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soybean meal
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acids (using fish meal and soybean reference)
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Figure 2. Differences between insect orders studied in the percentage of essential amino
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fish meal and soybean reference)
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Figura 3. Differences between insect orders studied in the percentage of fat acids (using
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soybean meal
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Figure 4. Cluster analysis of fatty acids similarity among insect species, fish meal and
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Table 1. Order, stage of developement and origin of the species of insect analised
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Table 2. Proximate analysis (% dry matter) of selected insects, fish meal and soybean meal
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Fish meal Soybean meal
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CP % 65.7 ± 1.3 34.6 ± 0.3 58.4 ± 0.4 53.5 ± 0.4 48.3 ± 0.9 61.8 ± 0.3 46.8 ± 1.1 40.9 ± 0.9 36.2 ± 0.3 40.7 ± 0.4 53.5 ± 4.4 59.0 ± 1.5 46.9 ± 4.1 40.1 ± 0.4 46.3 ± 0.6 56.0 ± 2.0 73.1 ± 3.3 66.0 ± 5.0 64.9 ± 0.5 74.4 ± 1.0 58.5 ± 0.5 73.0 ± 0.8 50.4 ± 0.2
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EE % 15.9 ± 1.4 11.8 ± 1.5 30.1 ± 0.7 38.0 ± 0.3 20.1 ± 0.7 27.0 ± 3.2 16.5 ± 0.0 5.8 ± 0.6 18.0 ± 1.6 15.6 ± 0.1 28.4 ± 1.5 26.6 ± 1.0 31.3 ± 1.6 33.7 ± 0.7 28.3 ± 0.6 23.6 ± 0.3 15.9 ± 0.2 17.6 ± 0.2 23.2 ± 0.6 8.8 ± 0.0 29.9 ± 0.5 8.2 ± 0.0 3.0 ± 0.0
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ASH % 7.8 ± 0.2 6.6 ± 0.6 3.5 ± 0.2 2.5 ± 0.3 8.0 ± 0.1 7.2 ± 0.1 6.1 ± 0.1 13.9 ± 0.4 9.3 ± 0.3 19.7 ± 0.1 4.9 ± 0.9 4.9 ± 0.2 6.5 ± 1.5 8.4 ± 2.9 3.9 ± 0.1 8.8 ± 0.1 5.6 ± 0.0 3.7 ± 0.1 4.8 ± 0.1 5.1 ± 0.1 4.0 ± 0.0 18.0 ± 0.2 7.8 ± 0.0
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PeA C RfL C TmL C ZmL C CvL D CmL D CmP D EtL D HiL D HiP D LsL D LsP D MdL D MdP D PtL D PtP D AdA O AaA O GaA O HlA O LmA O FM SM
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Specie Phyllognathus excavatus Rhynchophorus ferrugineus Tenebrio mollitor Zophoba morio Calliphora vicina Chrysomya megacephala Chrysomya megacephala Eristalis tenax Hermetia illucens Hermetia illucens Lucilia sericata Lucilia sericata Musca domestica Musca domestica Protophormia terraenovae Protophormia terraenovae Acheta domestica Anacridium aegyptium Gryllus assimilis Heteracris littoralis Locusta migratoria
NFE % 10.6 ± 0.1 47.0 ± 1.3 8.0 ± 0.2 6.0 ± 1.1 23.6 ± 0.1 4.0 ± 3.4 30.6 ± 1.1 39.4 ± 1.1 36.5 ± 1.0 24.0 ± 0.7 13.2 ± 4.6 9.5 ± 0.1 15.3 ± 4.0 17.8 ± 0.3 21.5 ± 0.1 11.6 ± 2.2 5.4 ± 0.3 12.7 ± 4.8 7.0 ± 0.3 11.7 ± 1.0 7.6 ± 0.1 0.8 ± 0.7 38.8 ± 0.3
Values are means ± SD of triplicate determinations. EE - Crude fat. CP - Crude protein. NFE -
Nitrogen-free extract
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Table 3. Amino acid (as a percentage of total amino acids) of selected insects, fish meal and soybean meal
FM SM
PHE 4.17 5.81 4.29 5.00 7.12 7.17 6.73 6.64 6.88 6.22 7.42 7.02 7.01 6.86 8.10 7.15 4.23 5.00 4.10 3.63 3.84 5.38 5.79
PRO 5.80 7.38 7.17 5.62 4.71 4.56 4.76 5.06 6.16 5.56 4.95 4.83 5.33 5.37 4.91 4.98 5.84 7.21 6.20 6.75 7.46 4.76 4.99
THR 4.10 4.00 4.49 4.33 4.86 4.51 5.02 5.02 5.39 4.95 5.38 4.60 4.87 5.28 4.78 4.83 4.10 4.49 4.11 3.90 4.28 6.26 4.17
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MET 1.42 0.45 0.64 0.76 2.16 2.22 2.76 2.37 1.50 3.26 3.36 3.08 3.00 3.44 2.30 2.55 1.49 2.36 1.10 1.02 0.54 2.93 1.01
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LYS 6.34 6.18 6.03 5.82 7.99 8.53 7.87 8.45 7.60 7.31 7.66 7.91 8.36 7.57 8.23 7.89 6.16 5.73 6.46 6.01 6.33 8.78 6.34
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LEU 7.59 6.63 8.65 8.25 6.69 6.89 6.96 7.62 6.87 6.83 6.43 6.96 6.75 6.57 6.29 6.77 8.69 7.28 8.06 8.88 8.31 7.81 8.01
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ILE 5.71 5.85 5.87 6.36 5.09 4.85 5.23 6.16 5.76 5.34 5.05 5.10 4.89 5.20 5.20 5.08 5.31 5.16 5.05 5.34 5.27 5.04 5.47
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HIS 4.19 3.62 3.64 3.87 5.02 5.08 5.20 4.22 5.29 5.16 5.12 5.18 4.68 5.17 5.48 5.35 2.93 4.04 3.03 2.69 2.98 7.86 3.28
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ARG 5.76 4.82 6.14 5.72 8.57 7.83 8.99 7.69 8.24 8.05 8.84 7.67 6.83 8.76 7.49 7.71 8.53 9.63 9.23 7.93 7.58 7.42 8.03
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PeA C RfL C TmL C ZmL C CvL D CmL D CmP D EtL D HiL D HiP D LsL D LsP D MdL D MdP D PtL D PtP D AdA O AaA O GaA O HlA O LmA O
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Specie
TYR 4.26 9.84 4.18 6.28 6.36 6.68 6.02 5.21 6.35 7.14 6.64 6.29 5.79 5.91 7.43 7.10 4.91 5.75 4.31 4.51 4.48 3.91 2.93
VAL 7.15 6.69 7.61 7.55 5.93 5.77 6.14 6.52 6.31 6.34 5.88 6.03 6.08 6.08 5.99 6.05 6.99 6.64 6.60 7.48 7.01 5.56 5.45
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Table 4. Fatty acid content (as a percentage of total fatty acid) of selected insects, fish meal and soybean meal
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18:3 n3
18:4 n3
20:5 n3
22:5 n3
22:6 n3
PeA C
0.9 ± 0.1
0
0
0
0
Other 1.5 ± 2.1
28.7 ± 3.0
Satura.
Monoun.
Ω -6
Ω-3
Polyuns.
58.1 ± 0.5
11.8 ± 0.4
0
11.8 ± 0.4
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Specie
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Table 4. Fatty acid content (as a percentage of total fatty acid) of selected insects, fish meal and soybean meal (cont.)
1.9 ± 0.1
0
0
0
0
0.1 ± 0.0
42.5 ± 0.4
44.4 ± 0.0
13.0 ± 0.4
0
13.0 ± 0.4
1.1 ± 0.0
0
0
0
0
0.6 ± 0.0
22.2 ± 0.1
45.8 ± 0.1
31.5 ± 0.1
0
31.5 ± 0.1
ZmL C CvL D
1.4 ± 0.0
0
0
0
0
1.6 ± 0.1
38.8 ± 0.2
35.7 ± 0.4
24.0 ± 0.0
0
24.0 ± 0.0
0.2 ± 0.3
1.3 ± 0.1
1.4 ± 0.0
0.7 ± 0.1
0.4 ± 0.0
2.9 ± 0.0
28.5 ± 0.2
40.7 ± 0.4
24.1 ± 0.1
3.9 ± 0.3
28.0 ± 0.1
CmL D
0.6 ± 0.0
0.4 ± 0.0
1.3 ± 0.1
0.6 ± 0.1
0
2.3 ± 0.6
35.9 ± 1.2
30.6 ± 0.1
28.5 ± 0.5
2.9 ± 0.4
31.3 ± 0.7
CmP D
0.5 ± 0.0
0.7 ± 0.1
1.5 ± 0.1
0.4 ± 0.0
0.2 ± 0.2
3.0 ± 1.1
35.4 ± 0.6
35.4 ± 0.7
23.0 ± 0.3
3.2 ± 0.4
26.2 ± 0.1
6.5 ± 1.2
41.7 ± 1.4
50.3 ± 0.2
1.6 ± 0.0
0
1.6 ± 0.0
0.1 ± 0.2
67.1 ± 0.6
16.9 ± 0.2
15.2 ± 0.4
0.7 ± 0.1
15.9 ± 0.6
0.6 ± 0.0
65.8 ± 0.1
32.6 ± 0.1
1.1 ± 0.0
0
1.1 ± 0.0
0
0
0
0
0.7 ± 0.1
0
0
0
0
HiP D
0
0
0
0
0
LsL D
0.4 ± 0.0
0.2 ± 0.0
0.1 ± 0.0
0
LsP D
0.3 ± 0.0
0
0.4 ± 0.0
0
MdL D
0.3 ± 0.0
0.1 ± 0.0
0.1 ± 0.1
0
MdP D
0.3 ± 0.0
0.2 ± 0.0
0.1 ± 0.1
0
PtL D
0.8 ± 0.0
1.1 ± 0.1
0.8 ± 0.0
0
PtP D
0
0.9 ± 0.0
1.8 ± 0.0
0
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AaA O
17.9 ± 0.4
0
0
GaA O
1.8 ± 0.0
0
0.7 ± 0.1
HlA O
19.4 ± 0.1
LmA O
4.7 ± 0.1
FM
0.2 ± 0.3
SM
6.9 ± 0.2
7.8 ± 0.3
27.8 ± 0.1
55.0 ± 0.1
8.8 ± 0.4
0.7 ± 0.0
9.5 ± 0.4
6.0 ± 0.0
28.8 ± 0.4
54.3 ± 0.2
10.3 ± 0.1
0.7 ± 0.0
11.0 ± 0.1
0
7.2 ± 0.0
32.6 ± 0.1
52.7 ± 0.2
7.1 ± 0.0
0.5 ± 0.1
7.6 ± 0.1
0
8.5 ± 0.1
30.0 ± 1.1
54.1 ± 0.8
7.0 ± 0.4
0.6 ± 0.1
7.5 ± 0.4
0
6.3 ± 0.1
27.1 ± 0.2
44.8 ± 0.1
19.2 ± 0.1
2.7 ± 0.1
21.9 ± 0.2
0
8.3 ± 1.9
26.6 ± 1.6
43.5 ± 0.1
19.2 ± 0.2
2.5 ± 0.0
21.7 ± 0.2
0
0
1.3 ± 0.1
34.2 ± 0.1
21.3 ± 0.1
43.2 ± 0.1
0
43.2 ± 0.1
0
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1.0 ± 0.0
30.3 ± 1.7
38.7 ± 1.0
30.0 ± 0.7
0
30.0 ± 0.7
0
1.0 ± 0.7
34.0 ± 0.6
27.5 ± 0.4
36.9 ± 0.2
0.7 ± 0.1
37.5 ± 0.3
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0.8 ± 0.0
0
0
0
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0.4 ± 0.0
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27.7 ± 0.6
27.6 ± 0.3
41.7 ± 0.1
0.4 ± 0.0
42.1 ± 0.1
0
0
0
0
0.1 ± 0.1
36.4 ± 0.1
47.8 ± 0.5
15.9 ± 0.4
0
15.9 ± 0.4
1.9 ± 0.0
14.1 ± 0.2
2.7 ± 0.1
16.1 ± 0.1
6.0 ± 0.4
36.1 ± 1.1
20.6 ± 0.7
2.7 ± 0.2
34.7 ± 0.2
37.3 ± 0.0
0
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0
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5.6 ± 0.8
24.0 ± 1.9
15.1 ± 0.3
55.4 ± 0.8
0
55.4 ± 0.8
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AdA O
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Highlights The aim of this paper is to show the insects as a sustainable alternative source to fish meal and soy meal We have evaluated nutritionally (protein, lipid, aminoacids, fatty acids) several insects. We have classified the insects regarding the similarity to the fish meal and soy meal
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S0044-8486(13)00679-0 doi: 10.1016/j.aquaculture.2013.12.024 AQUA 630970
To appear in:
Aquaculture
Received date: Revised date: Accepted date:
18 June 2013 11 December 2013 12 December 2013
Please cite this article as: Barroso, Fernando G., de Haro, Carolina, S´ anchez-Muros, Mar´ıa-Jos´e, Venegas, Elena, Mart´ınez-S´anchez, Anabel, P´erez-Ba˜ n´on, Celeste, The potential of various insect species for use as food for fish, Aquaculture (2013), doi: 10.1016/j.aquaculture.2013.12.024
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ACCEPTED MANUSCRIPT
The potential of various insect species for use as food for fish
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Fernando G. Barroso 1*, Carolina de Haro 1, María-José Sánchez-Muros 1, Elena Venegas 2, Anabel Martínez-Sánchez3 & Celeste Pérez-Bañón3
Department of Applied Biology, University of Almería, Almería, Spain.
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Food Technology Division, University of Almería, Almería, Spain.
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Instituto Universitario CIBIO, University of Alicante, Alicante, Spain.
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* Corresponding author:
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Fernando G. Barroso
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Department of Applied Biology,
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E.S.I. University of Almería,
Carretera de Sacramento s/n, 04120 Almería, Spain
Fax: +34950015476
Phone: +34950015918
e-mail: [email protected]
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ACCEPTED MANUSCRIPT Abstract Due to the expansion of aquaculture and the limited resources available from the sea, it
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is necessary to find substitutes for fish meal for use in aquaculture. We believe that the
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use of insect meals as an alternative source of animal protein may be an option. To use insects for this purpose, it is necessary to determine the nutritive characteristics of these
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insects. To determinate the potential of insects as a substitute for fish meal in fish food
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used in aquaculture, we examined 16 different species, 5 of then as different stage of development, of the orders Coleoptera (4), Diptera (7) and Orthoptera (5). The insect
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analysed have a higher proportion of fat and less protein than fish meal. With the exceptions of histidine, threonine and lysine, the insects present an amino acids profile
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similar to fish meal, being Diptera b the most similar group to fish meal. However, the
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fatty acid content of insects is very different from that of fish meal that is rich in n-3,
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specially 14% EPA, 16% DHA, which are practically absent in insects, how have higher ratios of omega 6 and monounsaturated fat.
KEY WORDS: fish nutrition, alternative feed, insects meal, fish meal, amino acids, fatty acid.
1. Introduction Fish have been a key source of food for humans (Ayoola, 2010), but the global catch of wild fish declined approximately 3% from 2004 to 2009 at a rate of 0.5% per year (FAO, 2010). Currently, aquaculture is playing an essential role in the seafood market, meeting the demand for fish that cannot be met with the wild catch. As a consequence,
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ACCEPTED MANUSCRIPT in recent years (from 2004 to 2009), aquaculture production has grown by 32%, a growth rate of approximately 5.6% per year (FAO, 2010).
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Fishmeal is one of the major components of the feed used in aquaculture. It is
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generally added to animal diets to increase feed efficiency and animal growth through better feed palatability; it also enhances the uptake, digestion, and absorption of
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nutrients (Mile and Chapman, 2006). It is estimated that approximately 30% of the total
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fish catch is converted to fish meal and fish oil for use in animal and fish feeds (Ogunji et al., 2006).
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The percentage of fish meal that is used for aquaculture feeds has increased from 10% in 1988 to approximately 45% in 2002. The increasing global demand for and
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decreasing availability of fish meal has led to sharp increases in the price of fish meal,
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and hence, the cost of aquaculture production has increased as well (Ayoola, 2010). The
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price of producing fish through aquaculture has risen from US $600/metric ton in 2005 to US $2000/metric ton in June 2010, and this trend is likely to continue (International Monetary Fund, 2010).
The present shortage of fish meal motivates researchers to seek new protein sources with nutritional values similar to fish meal, in particular those with similar contents of the essential amino acids, phospholipids, and fatty acids (docosahexaenoic acid and eicosapentaenoic acid) that promote optimum development, growth, and reproduction (Ayoola, 2010), which would allow aquaculture production to remain economically and environmentally sustainable over the long term. From vegetable sources, soybean meal is the best available vegetable protein source in terms of protein content and EAA profile. However, it is potentially limiting in sulphur-
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ACCEPTED MANUSCRIPT containing amino acids (methionine and cysteine) and contains some antinutrient substances such as trypsin inhibitor, haemagglutinin, and antivitamins (Tacon, 1993).
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Regarding sources of animal origin mostly of them are forbidden by prescription of
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food security, which have made more urgent the search for alternatives to fish meal in aquaculture diets (Ogunji, 2004).
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Edible insects are a natural renewable resource used as food by humans (Ramos-
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Elorduy and Conconi, 1994). Since ancient times, insects have been one alternative protein source used to compensate for the periodic or seasonal scarcity of other sources
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(Ramos‐Elorduy, 1997). The most existing studies have focused on the insects that have played an important role in human nutrition in Africa, Asia, and Latin America. Thus,
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we can highlight the studies conducted in Nigeria (Akinnawo and Ketiku, 2000; Banjo
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et al., 2006), Mexico (Ramos-Elorduy and Conconi, 1994; Ramos-Elorduy et al., 1997;
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Ramos-Elorduy et al., 2006; Ramos‐Elorduy, 1997), Thailand (Yhoung‐Aree et al., 1997) and Zaire (Kitsa, 1989). Although insects began to be evaluated as a potential foodstuff for animals 40 years ago (Calvert et al., 1969; Hale, 1973; Ichhponani and Malik, 1971; Newton et al., 1977; Phelps et al., 1975; Teotia and Miller, 1974), the incorporation of insects into fish feed has not received much attention until recently (Ogunji et al., 2006). In the last 10 years there have been several studies of feeding experiments performed in vivo with diets based on insect meal in Clarias anguillaris (Achionye-Nzeh and Ngwudo, 2003), Clarias gariepinus (Alegbeleye et al., 2012; Aniebo et al., 2011; Fasakin et al., 2003), Orcorhychus mykiss (Sealey et al., 2011; St-Hilaire et al., 2007; ) Oreochromis niloticus (de Haro et al., 2011a; de Haro et al., 2011b; de Haro et al., 2011c; de Haro et al., 2011d; de Haro et al., 2011e; Ogunji et al., 2006; Ogunji et al., 2008) and Psetta
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ACCEPTED MANUSCRIPT maxima (Kroeckel et al., 2012). In general, percentages of substitution higher than 30% decreased the growth depending on the fish and insect species.
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From the nutritional point of view, depending on species and/or stage, insects are rich in
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protein and lipids, nevertheless the presence of chitin a priori indicates a negative characteristic. However, chitin also is present in crustacean, which are widely consumed
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by fish.
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To evaluate the potential of insects, it is necessary to consider other advantages such as environmental benefits; the insects can be fed with waste generated by humans, having
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an important role in recycling materials in the terrestrial biosphere (Katayama et al., 2008). The great diversity of insect species (70-75% of animal species), from different
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ecosystems, with different diets and stages of development (larval, pupa, ninpha or
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imago) provokes a huge variability in body composition.
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The aims of this study are: (1) to determine the nutritive valour of the most frequent rearing insects and some of the common species of Almeria (Spain), and (2) to establish the most similar to fish meal.
2. Materials and methods 2.1. Sampling
The insects and stages of its development (larval, pupa, ninpha or imago) studied were chosen according to the following criteria: easy to rearing, stage with mayor biomass and low exoskeleton. The wild species were chosen in terms of their availability in the environment. The insects used in the study were obtained from pet shops (captivity*), reared by the research group "Bionomía, Sistemática e Investigación Aplicada de insectos dípteros e
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ACCEPTED MANUSCRIPT himenópteros" of the University of Alicante (captivity **) or captured in the field close
belong to three orders, Diptera, Orthoptera and Coleoptera.
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to our environment (wild-rearing). Table 1 summarises the insects studied, which
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The nutritional contents of insects were compared with those of fish meal and soybean
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meal because they are the most common ingredients used in aquafeed production.
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2.2. Analytical methods 2.2.1. Determination of proximate composition
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The nutritional values obtained were derived from three replicate samples for each species of insecta, fish meal and soy meal. Moisture, crude protein, total lipids and ash
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were determined using AOAC (2005) techniques.
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The insects were sacrificed by freezing (Finke et al., 1989). All samples, insects,
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fishmeal and soy meal were lyophilised (Cryodos, Ima-Telstar, Terrassa, Spain), and ground and freezing until to be analized. Total nitrogen (N) was determined using the Kjeldahl procedure, and crude protein was estimated as N x 6.25. Crude lipid was determined following the Soxhlet extraction of dried samples with petroleum ether. Moisture was determined after oven drying the samples at 105 ºC to a constant weight. The ash was determined eliminating the organic matter at 500ºC during 12h.
2.2.2. Determination of amino acid profile The amino acid profile was determined after hydrolysing the sample with 6 N HCl for 22 h at 110 ºC, followed by a sequence of filtering, derivatisation, and separation in a gas chromatograph. Tryptophan was not determined.
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ACCEPTED MANUSCRIPT 2.2.3. Analysis of fatty acids For the FA analyses, all samples were transmethylated following the method of Lepage
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and Roy (1984) with the minor modifications of Venegas-Venegas et al. (2011): for
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each sample, 1 ml of freshly prepared transesterification reagents (methanol/acetyl chloride, 20:1, v/v) was added to 50 mg of freeze-dried insect meal in a glass tube along
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with 100 µl of a solution of internal standard (heptadecanoic acid 17:0, 10 mg/ml). The
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tubes were shaken and then placed in a hot block (100 ºC, 30 min). Next, the mixture was cooled to room temperature, and 1 ml of distilled water was added to each tube.
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The samples were shaken again and centrifuged (3,000 rpm, 3 min). The upper hexane phase was collected for GLC analysis.
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The resulting FA methyl esters (FAMEs) were analysed in a Focus GLC (Thermo
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Electron, Cambridge, UK) equipped with a flame injection detector (FID) and an
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Omegawax 250 capillary column (30 m 9 0.25 mm i. d. 9 0.25 lm film thickness; Supelco, Bellefonte, PA, USA). The temperature program was 1 min at 90 ºC, heating to 200 ºC at a rate of 10 ºC/min, constant temperature at 200 ºC (3 min), heating to 260 ºC at a rate of 6 C/min and constant temperature at 260 ºC (5 min). The injector temperature was 250 ºC with a split ratio of 50:1. The injection volume was 4 µl. The detector temperature was 260 ºC. Nitrogen was used as the carrier gas (1 ml/min). Total saturated, monounsaturated, polyunsaturated, n-3 and n-6 fatty acids were calculated as the sums of saturated fatty acids (ΣSFA), monounsaturated fatty acids (ΣMUFA), polyunsaturated fatty acids (ΣPUFA), n-3 and n-6 fatty acids, respectively.
2.3. Data analysis
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ACCEPTED MANUSCRIPT To better understand the applied value of the study’s results, in addition to a descriptive approach to the nutritional value of the insects, the compositions of the insect meals
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were compared with the compositions of fish meal and soybean meal.
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To determine the similarity between the compositions of fish meal, soybean meal and the different species of insects, a hierarchical cluster analysis was used.
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Clustering is a multivariate technique of grouping together rows that share similar
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values. Cluster analysis is an exploratory data analysis technique for solving classification problems. Its purpose is to sort the different food sources (fish meal,
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soybean meal and insect meals) into groups (clusters) so that the degree of association/similarity between members of the same cluster is stronger than the degree
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of association/similarity between members of different clusters. Each cluster is
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described as the class to which its members belong. This analysis provides a
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dendrogram grouping the meals according to the distance between them. The similarities were evaluated using hierarchical cluster analysis in the Past (Paleontological Statistics, Version 2.17). The analysis was performed using hierarchical option clusters based on Ward’s method (Ward, 1963) with standardised data. Bootstrapping has previously been used to obtain confidence intervals. Specifically, there were two cluster analyses based on different characteristics: •
The proportion of essential amino acids.
•
The proportion of fatty acids
We observed that the cluster analysis classified different insect species according to their taxonomic order. Therefore, to analyse the differences between orders, we grouped the data by order regardless of the species. An analysis of variance with the taxonomic order as main factor was performed with the JMP (Version 9.0.0) statistical package
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ACCEPTED MANUSCRIPT (SAS Institute, Inc.) to evaluate the relationship between the compositions of amino acids and fatty acids. Significant differences in the mean response were determined with
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the Tukey-Kramer HSD test.
3. Results
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The proximate analysis of the insect samples is summarised in table 2. As expected,
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most insect species analysed have a high proportion of protein, between 40 and 60% in similar to soy meal levels (50% CP) and lower than fishmeal (73.0% CP). The lowest
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levels were measured in Rhynchophorus ferrugineus (35%) and Hermetia illucens larvae (36%). The protein content of Orthoptera (73% CP) is similar to fish meal
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(73.0%), particularly Heteracris littoralis (74%) and Acheta domestica (73%).
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The lipid levels in fish meal (8.2%) and soy meal (3.0%) are lower than in insects. The
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lipid contents of most of the insects reached 20%. The highest values were obtained for Locusta migratoria, Musca domestica larvae and Tenebrio mollitor with 30% lipids and Zophoba morio with 38.0% lipids. These insects contain between 5 and 20% of nitrogen free extract (NFE). Only three of the species had nitrogen free extracts above 35%. NFE involve carbohydrates, sugars, starches, fiber and chitin (primarily a nitrogen-containing polysaccharide). Regarding ash, fish meal contains a higher percentage (18.0%) than insect meals (except for Hermetia pupae, 19.7%) or soybean meal. The amino acid analyses are summarised in table 3. In general, the amino acid patterns were quite different among species. The dendrogram in figure 1 illustrates the similarity of the essential amino acid profiles of the insects, fish meal and soybean meal. As shown in figure 1, two major groups or
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ACCEPTED MANUSCRIPT cluster were formed; (1) fish meal and Diptera, (2) Orthoptera, soybean meal and Coleoptera. Thus, Diptera appears to be the most similar to fish meal in terms of its
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amino acid composition, especially the larvae of Hermetia, Musca and Eristalis,
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whereas Coleoptera is the most different from fish meal in amino acid composition. Soybean meal is closer to Orthoptera in the dendrogram. These results verified the
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relationship between similarity in amino acid composition and insect taxonomy, as each
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order was grouped into a different cluster.
Figure 2 presents the significant differences in the proportion of essential amino acids
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among the three orders of insects, fish meal and soybean meal for all essential amino acids. Relative to fish meal, the insect meals are deficient in the amino acids histidine,
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lysine and threonine, but better in lysine methionine and tyrosine than soy meal. Diptera
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shows a histidine, lysine and threonine proportion similar to fish meal (figure 2 and
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table 3). Diptera also has a similar proportion of methionine and a higher proportion of phenylalanine compared to fish meal and soy meal. However, Diptera exhibits a relative deficit in leucine that does not occur in Orthoptera or Coleoptera. The percentages of tyrosine and valine were higher in all analysed insects than in fish meal. These data reveal that Diptera has similar amino acids profile to fish meal. Ortopthera and Coleoptera, although not so similar to fish meal, have better amino acids profile than soy meal. Table 4 and figure 3 show fatty acids composition of insect fish meal and soy meal. The most notable difference was the higher percentage of n-3 fatty acid in fish meal, up to 37% of the total fatty acids, containing 14% EPA (20:5 n3) and 16% DHA (22:6 n3). In contrast, the insects have a much lower proportion of n-3, while soy mal a rich in n-6. Some species of Orthoptera show a higher proportion (Hetteracris littoralis, 19.8% and
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ACCEPTED MANUSCRIPT Anacridium aegyptium, 17.9%), but the n-3 in those species consists primarily of aclinolenic acid (ALA 18:3 n3). Chrysomya megacephala and Calliphora vicina (Diptera)
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only contain EPA, but it does not exceed 1.5% of total FAs, a low percentage compared
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with the fish meal values.
The saturated fatty acid compositions of the insect meals are similar to fish meal;
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nevertheless, insects have nearly twice monounsaturated fatty acid than fish meal and
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soy meal . The difference in n-6 is much larger; fish meal contains only 2.5% n-6, and soy meal 55,4%, whereas insects have a intermediate proportion, in some species
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reaching over than 30% (e.g., Tenebrio mollitor, Gryllus assimilis and Acheta domestica). Eristalis tenax is the only studied species that had a proportion (1.9%) of n-
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The variability of fatty acid profiles among species (figure 3) was greater than the
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variation in essential amino acids, with significant differences in only some fatty acids. The Orthoptera exhibited a higher proportion of n-3, especially for ALA 18:3 n3. No differences between groups for n-6 acids were observed except for the lower proportion of linoleic acid (18:2 n6) in Diptera compared to Orthoptera. The dendrogram (figure 4) illustrates the similarities of the fatty acid compositions among insects, fish meal and soybean meal. The results of the cluster analysis show several groups and the distances are higher than the amino acids. Farthest group is composed of fish meal, Eristalis tenax and Hermetia ilucens larvae. Soybean meal and the other insects (Diptera, Coleoptera and Orthoptera) are mixed in more distanced groups.
4. Discussion
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ACCEPTED MANUSCRIPT The results obtained of nutrients composition of insect show differences regarding fishmeal and among species and/or stage.
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4.1. Proximal composition
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Most of the insects species analysed have a high proportion of protein, similar to the levels of soy meal but lower than fish meal. In general, the order Orthoptera exhibits a
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higher CP, between 60 and 70%, but it must be taken into account that all samples of
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this order are adults with more chitin (and chitin nitrogen) and less fat that enhances CP levels. In Diptera, the levels of CP range from 40 to 50%, with higher values for the
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larvae of Chrysomya megacephala, pupae of Lucilia sericata and pupae of Protophormia terraenova (50-62%). The CP values vary among published works, the
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CP value obtained for larvae of Musca domestica was 46.9%, similar to that reported by
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Ogunji et al. (2008), 47.1%, whereas other authors have obtained more diverse results,
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including 37.5% (Aniebo and Owen, 2010) and 56.8% (Bernard et al., 1997). In pupae of Musca domestica, Bernard et al. (1997) obtained a higher proportion, 58.3%, than that observed in this experiment, 40.1%. The percentage of CP obtained in larvae of Hermetia illucens (36.2%) is similar to those obtained by Sheppard (2002) (37.8%) and Arango et al. (2004) (37%) and slightly lower than the value of 40.6% described by Newton et al. (1977). The CP content obtained in Eristalis tenax, 40.9%, is similar to that observed by Ramos-Elorduy et al. (1998) in Eristalis sp. (40.7%). In the order Coleoptera, the species with the least CP is Rhynchophorus ferrugineus (35%). The larvae of the genus Rhynchophorus are not very protein-rich; Banjo et al. (2006) reported that Rhynchophorus phoenicis was only 28.4% protein, and Cerda et al. (1999) reported 25.8% protein for Rhynchophorus palmarum. In Tenebrio mollitor larvae, the observed CP was 58.4%, and in Zophoba morio, 53.5%. Both the Tenebrio
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ACCEPTED MANUSCRIPT and Zophoba values are slightly higher than those observed by other authors: between 47 and 53% for Tenebrio mollitor (Bernard et al., 1997; Finke, 2002; 2007; Ramos-
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Elorduy et al., 2006) and 46.8% for Zophoba morio (Finke, 2002).
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These differences may be due to the phase of development of the insect, variations in dietary habits between populations, the method of processing or differences in ecotypes
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(Banjo et al., 2006; Fasakin et al., 2003; Teguia and Beynen, 2005), and this makes it
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difficult to draw solid conclusions related to differences between orders. Nevertheless the results of this study indicate that the protein content of insects is
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generally less than that of fish meal and it is similar to soybean meal. There are substantial differences between the protein contents reported by different authors, even
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for the same species of insect. This must be investigated to know the species and stages
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appropriate to be used in fish feeding.
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On the other hand, an additional problem to CP determination in insect is the nitrogen contained within nacetylglucosamine, a subunit of the chitin polymer, digestibly unavailable (Finke et al., 1989). Finke (2002) suggests that the amount of nitrogen contained in chitin is relatively small, and thus, nitrogen content may provide a reasonable estimate of total protein in most invertebrates. The fat content of insects is highly variable and it seems to vary between stages, usually ranging between 15 and 30%. Zophoba morio is the species with the highest proportion of fat (38%) of all of the analysed species; this value is slightly lower than that obtained by Finke (2002), 42%. The larvae of coleopterans generally have a large amount of fat, often exceeding 25%, the high proportion of fat was recorded for Tenebrio mollitor (30%), lower than levels (38 to 43%) reported by other authors (Finke, 2002; 2007; Ramos-Elorduy et al., 2006). A lower proportion was obtained in this study for the
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ACCEPTED MANUSCRIPT larvae of Eristalis tenax (5.8%), lower than that obtained by Ramos-Elorduy et al. (1998) in Eristalis sp. (11.9%). Fat levels in both, larvae (31.3%) and pupae (33.7%) of
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Musca domestica are greater than those observed by other authors: from 13.5 to 25% in
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larvae (Aniebo and Owen, 2010; Bernard et al., 1997; Ogunji et al., 2006, 2008; Sheppard, 2002) and 15.8% in pupae (Bernard et al., 1997). As reported by Arango et
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al. (2004), the percentage obtained for the larvae of Hermetia illucens, 18%, is lower
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than the values observed by Newton et al. (1977) and Sheppard (2002), both greater than 30%.
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The stage of development may also affect the fat content. In general, the larval stages contained significantly more fat than adults (Barker et al., 1998). In Acheta domestica
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varies between 14 and 22% (Bernard et al., 1997; Finke, 2002; 2007) at different stages
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of development. Probably the differences in lipids among the same species, reported by
age.
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different authors, could be related not only to the stage of development but also to the
Diet appears to be another factor that influences the fat content of insects. A comparison of the fat content of the wild orthopteran Heteracris littoralis, at 8.2%, with captivebred orthopterans (Acheta domestica, Gryllus assimilis and Locusta migratoria), with a higher proportion of fat, suggests that diet could affect lipid content. As it occurs with CP, the lipids content varies enormously. Because of this, it is difficult to obtaine a clear conclusion regarding relation between lipids content and taxon, stage or feeding. Supplementary studies are needed. On the other hand, the lipid levels in fish meal and soy meal are lower than in insects, which could complicate the use of insect meals in fishfeeds.
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ACCEPTED MANUSCRIPT The ash contents obtained for all analysed insects were less than that of fish meal (18.0%), with the exception of Hermetia illucens pupae (19.7%). Arango et al. (2004)
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(17.5%) and Newton et al. (1977) also found elevated levels (14.6%) of ash in this
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species.
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4.2. Amino acid profile
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A balanced essential amino acid (EAA) profile is one of the characteristics that define the quality of protein. The development of commercial aquatic feeds has traditionally
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been based on FM as the primary protein source due to its high protein content and balanced essential amino acid profile (Nguyen et al., 2009) with high level of digestible
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essential amino acids such as lysine, methionine and leucine, which are often deficient
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in grains, the typical base for most animal feeds (Hall, 1992; Keller, 1990). Currently,
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the specific amino acid composition of most insect species is unknown. In this work the amino acids profile of different species and stages have been studied. The results show that the profile of amino acids is related to the taxonomic group (figure 1) and Diptera are most similar, in terms of essential and limiting amino acids, to fish meal (Fig. 2). This group has a similar proportion of methionine to fish meal and higher values of histidine, lysine and threonine. Orthoptera and Coleoptera have a higher proportion of leucine. These results are consistent with several studies in which low levels of methionine have been observed in Orthoptera (Bernard et al., 1997; Finke, 2002; 2007) and Coleoptera (Cerda et al., 1999; Finke, 2002; 2007; Ramos-Elorduy et al., 2006) and high levels in Diptera (Newton et al., 1977; Ogunji et al., 2006). Regarding lysine, the proportions obtained for both Coleoptera and Orthoptera, 6%, are similar to those described by
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ACCEPTED MANUSCRIPT Finke (2002, 2007). In Diptera, the lysine level obtained in this work, 8%, coincided with the value previously reported by Newton et al. (1977).
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The proportions of arginine have been shown to be adequate for Diptera and Orthoptera;
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however, the Coleoptera are in deficit. These results differ somewhat from earlier studies, which found that the proportion of arginine in Orthopterans was approximately
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7% (Finke, 2002; 2007) and those in Diptera (Newton et al., 1977; Ogunji et al., 2006)
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and Coleoptera (Finke, 2002; 2007; Ramos-Elorduy et al., 2006) were approximately 5%.
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In terms of similarity to fish meal, the amino acid profile of the Diptera is superior to that of soybean meal, thus Diptera could be a better replacement than soybean meal in
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the formulation of fish feed. Although each species is deficient in some essential amino
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several species of insect.
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acids, it is possible to design a feedstuff that is balanced in amino acids by combining
4.3. Fatty acids
According to Ramos-Elorduy (2008) insects contain higher quantities of polyunsaturated fats (PUFAs) n-6 than fish meal, but lower than soy meal. As in soy, lower levels of polyunsaturated fats n-3 were observed in insect meal analysed regarding fish meal. Bukkens (1997) reported significant quantities of linoleic acid in the fatty acid content of all analysed species of insect. As observed by other researchers (Akinnawo and Ketiku, 2000; Beenakkers and Scheres, 1971; Ekpo and Onigbinde, 2007; Finke, 2002; 2007; Katayama et al., 2008), terrestrial insects do not contain EPA or DHA except for Chrysomya megacephala and Calliphora vicina, which have
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ACCEPTED MANUSCRIPT between 1.3 and 1.5% EPA, far lower proportions than fish meal, which contains up to 14 to 16% EPA.
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The lack of EPA and DHA in insects is one of the most important limitations to its use
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in seawater fish feed, since it involves a limit in their inclusion in aquafeed. Nevetheless there are evidences that the fatty acid profiles of insects most likely reflect
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the fatty acid composition of their food. In wild Hetteracris Littoralis and Anacridium
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aegyptium individuals that consume plants in the field have a high amount of a-linolenic acid (ALA, 18:3 n3), a precursor of the n-3 series, whereas three other species of
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orthopterans, bred in captivity and fed only with flour and bran cereals, present with lower ALA, 18:3 n3. Finke (2002) has achieved FA profiles virtually identical to those
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obtained in this work in Zophoba morio and Tenebrio mollitor, species bred in captivity.
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The fatty acid content of Locusta migratoria determined in this experiment is very
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different from that obtained by Beenakkers and Scheres (1971), who measured 26% oleic acid (18:1 n9) and 21% ALA (18:3 n3), whereas the results of this experiment show 45% oleic acid and 5% ALA. These differences could be related to diet; Locusta used in this experiment are from pet stores, whereas in the work of Beenakkers and Scheres (1971), Locusta were provided with a diet of reed, endive, or pear (supplemented with some reed). In addition, these authors obtained a fair correlation between the percentages on oleic and linoleic acid in the tri- and diglycerides of both diet and body fat. These results indicate that lipid quality could be manipulated by feeding, which amplifies the nutritive values of insects meal. On the other hand in this work we have not sampled any aquatic insect. Freshwater insects have a high proportion of EPA because they ingest freshwater algae, which generally contain ALA. EPA levels above 15% as have been observed by Bell et al. (1994).
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ACCEPTED MANUSCRIPT The data obtained in this experiment agree with Justi et al. ( 2003): fatty acids content of insects is more dependent on diet, in contrast with amino acids, more related to the
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taxon.
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The comparison with fish meal and soy meal reveals lower levels of n-3 and n-6 in
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insect meal.
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Finally, it is interesting to note the following considerations:
- The utilisation of insects has other advantages that cannot be ignored; insects do not
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compete with human resource and can be reared on by-products or human waste as efficient biotransformers to convert abundant low-cost organic wastes into animal
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biomass that is rich in proteins and suitable for use in animal nutrition (Ramos-Elorduy,
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1999; 2005). Regarding soybean production, which is largely oriented toward animal
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feed, insects require fewer natural resources (i.e., land, water, fertiliser) than soybean, the production of which could compete with human food production. - The current aquaculture production system, supported by fish meal, is clearly unsustainable. As reported by the FAO (2010), capture fisheries have been rather stagnant or even declining in some countries, particularly fisheries of wild marine fish stocks; 3% are under-exploited, 12% are moderately exploited, 53% are “fully exploited”, 28% are over-exploited, 3% are depleted and 1% is recovering from depletion. This situation, together with the increase in demand for fish, indicates that fish meal will become a more limiting ingredient in both production and price. Therefore, it is essential to obtain potential alternatives.
5. Conclusion
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ACCEPTED MANUSCRIPT The data obtained in this work indicate an adequate nutritional composition of the insect meals evaluated, for inclusion in fish food. The great variety of insect species, habitats,
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development stages, feeding habits and other characteristics most likely affects insect
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nutritional value and makes insect meal very interesting to study as an alternative to fish meal.
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Although insects generally present some characteristics that do not match with the fish
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meal, the amino acid profile of the Diptera shows that this group of insects could be a possible alternative protein source to be used in aquaculture. Nevertheless, more studies
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are needed to know the digestibility, chitin content and digestive effect, presence of toxic, meal treatments (such as degreasing), adequate mixtures of different insect
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species or to modify the nutritional value of insects by changing their diet or rearing
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condition. Insects can potentially play a fundamental role in animal nutrition, so further
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studies are required on their nutritional value in the context of their use in animal feed.
Acknowledgements
The authors are very grateful to the Consejería de Innovación y Ciencia, Junta de Andalucía (project AGR5273), Fondos Europeos de Desarrollo Regional (FEDER Funds) and Campus de Excelencia Internacional Agroalimentario y del Mar, Ministerio de Educación, by financial support.
References
19
ACCEPTED MANUSCRIPT Achionye-Nzeh, C.G., Ngwudo, O.S., 2003. Growth response of Clarias anguillaris
Bioscience Research Communications. 15, 221-223.
PT
fingerlings fed larvae of Musca domestica and soyabean diet in the laboratory.
RI
Akinnawo, O., Ketiku, A.O., 2000. Chemical composition and fatty acid profile of edible larva of Cirina forda (westwood). African Journal of Biomedical
SC
Research. 3, 93-96.
NU
Alegbeleye, W.O., Obasa, S.O., Olude, O.O., Otubu, K., Jimoh, W., 2012. Preliminary evaluation of the nutritive value of the variegated grasshopper (Zonocerus
MA
variegatus L.) for African catfish Clarias gariepinus (Burchell. 1822) fingerlings. Aquaculture Research. 43, 412-420.
D
Aniebo, A.O., Owen, O.J., 2010. Effects of age and method of drying on the proximate
TE
composition of housefly larvae (Musca domestica Linnaeus) meal (HFLM).
AC CE P
Pakistan Journal of Nutrition. 9, 485-487. Aniebo, A.O., Odukwe, C.A., Ebenebe, C.I., Ajuogu, P.K., Owen, O.J., Onu, P.N., 2011. Effect of housefly larvae (Musca domestica) Meal on the carcass and sensory qualities of the mud catfish, (Clarias gariepinus). Advances in Food and Energy Security. 1, 24-28. AOAC, 2005. Official methods of analysis of the Association of Official Analytical Chemists International, 18th edn ed. AOAC International, Gaitherburg, USA. Arango, G.P., Vergara, R.A., Mejía, H., 2004. Análisis composicional, microbiológico y digestibilidad de la proteína de la harina de larvas de Hermetia illuscens (diptera: Stratiomyiidae) en angelópolis-antioquia, Colombia. Revista Facultad Nacional de Agronomía, Medellín (online). 57, Available at: http://redalyc.uaemex.mx/src/inicio/ArtPdfRed.jsp?iCve=179914073009.
20
ACCEPTED MANUSCRIPT Ayoola, A.A., 2010. Replacement of fishmeal with alternative protein Source in aquaculture diets. Thesis Degree of Master of Science. Faculty of North
PT
Carolina State University. North Carolina, USA. Thesis Degree of Master of
RI
Science in Faculty of North Carolina State University.
Banjo, A.D., Lawal, O.A., Songonuga, E.A., 2006. The nutritional value of fourteen
SC
species of edible insects in southwestern Nigeria. African Journal of
NU
Biotechnology. 5, 298-301.
Barker, D., Marianne, P., Fitzpatrick, D., Dierenfeld, E.S., 1998. Nutrient composition
MA
of selected whole invertebrates. Zoo Biology. 17, 123–134. Beenakkers, A.M.T., Scheres, J.M.J.C., 1971. Dietary lipids and lipid composition of
D
the fat-body of locusta migratoria. Insect Biochemistry. 1, I25-I29.
TE
Bell, J.G., Ghioni, C., Sargent, J.R., 1994. Fatty acid compositions of 10 freshwater
AC CE P
invertebrates which are natural food organisms of Atlantic salmon parr (Salmo salar): a comparison with commercial diets. Aquaculture. 128, 301-313. Bernard, J.B., Allen, M.E., Ullrey, D.E., 1997. Feeding captive insectivorous animals: nutritional aspects of insects as food. Nutrition advisory group handbook. Fact sheet 003. Scientific Advisory Group to the American Zoo and Aquarium Association.
Bukkens, S.G.F., 1997. The nutritional value of edible insects. Ecology of Food and Nutrition. 36, 287–319. Calvert, C.C., Martin, R.D., Morgan, N.O., 1969. House fly pupae as food for poultry. Journal of Economic Entomology. 62, 938–939. Cerda, H., Martínez, R., N., B., Pizzoferrato, L., Hermoso, D., Paole, M., 1999. Cría, análisis nutricional y sensorial del picudo del cocotero Rhynchophorus
21
ACCEPTED MANUSCRIPT palmarum (coleoptera: curculionidae), insecto de la dieta tradicional indigena amazónica. Ecotropicos. 12, 25-32.
PT
de Haro, C., Barroso, F.G., Hidalgo, L., Sánchez-Muros, M.J., Venegas, E., 2011a.
RI
Effect in muscle composition of the inclusion of Tenebrio mollitor meal in tilapia feed (Oreochromis nilotica), Aquaculture Europe (Annual meeting of the
SC
European Aquaculture Society): Mediterranean aquaculture 2020, Rhodes,
NU
Greece.
de Haro, C., Barroso, F.G., Hidalgo, L., Sánchez-Muros, M.J., Uclés, F., 2011b.
MA
Nutritive adaptation in Nile tilapia (Oreochromis niloticus) to Tenebrio mollitor meal as protein source diet: effect in fish growth and performance, Aquaculture
D
Europe (Annual meeting of the European Aquaculture Society): Mediterranean
TE
aquaculture 2020, Rhodes, Greece.
AC CE P
de Haro, C., Sánchez-Muros, M.J., Rueda, M., Martínez, R., Barroso, F.G., 2011c. Efectos de la sustitución de la harina de pescado por harina de tenebrio (Tenebrio mollitor) en la alimentación para tilapia del Nilo (Oreochromis niloticus), XIII Congreso Nacional Acuicultura, Barcelona, Spain. de Haro, C., Barroso, F.G., Hidalgo, L., Sánchez-Muros, M.J., Férriz, M., 2011d. Evaluation of Tenebrio mollitor meal as a protein source in tilapia (Oreochromis nilotica) feed: nutritive utilization and effect in fish growth and performance, Aquaculture Europe (Annual meeting of the European Aquaculture Society): Mediterranean aquaculture 2020, Rhodes, Greece. de Haro, C., Barroso, F.G., Hidalgo, L., Rincón-Cervera, M., A., Sánchez-Muros, M.J., 2011e. Effect of adaptation to ingest a diet based in Tenebrio mollitor meal on body composition in Nile tilapia (Oreochromis niloticus), Aquaculture Europe
22
ACCEPTED MANUSCRIPT (Annual meeting of the European Aquaculture Society): Mediterranean aquaculture 2020, Rhodes, Greece.
PT
Ekpo, K.E., Onigbinde, A.O., 2007. Characterization of lipids in winged reproductives
RI
of the Termite Macrotermis bellicosus. Pakistan Journal of Nutrition. 6, 247251.
SC
FAO, 2010. The State of World Fisheries and Aquaculture 2010. FAO, Rome, Italy.
NU
Fasakin, E.A., Balogun, A.M., Ajayi, O.O., 2003. Evaluation of full-fat and defatted maggot meals in the feeding of clariid catfish Clarias gariepinus fingerlings.
MA
Aquaculture Research. 34, 733-738.
Finke, M.D., 2002. Complete nutrient composition of commercially raised invertebrates
D
used as food for insectivores. Zoo Biology. 21, 269-285.
TE
Finke, M.D., 2007. Estimate of chitin in raw whole insects. Zoo Biology. 26, 105-115.
AC CE P
Finke, M.D., DeFoliart, G.R., Benevenga, N.J., 1989. Use of a four-parameter logistic model to evaluate the quality of the protein from three insect species when fed to rats. The Journal of Nutrition. 119, 864-871. Hale, O.M., 1973. Dried Hermetia illucens larvae (Diptera: Stratiomyidae) as a feed additive for poultry. Journal of the Georgia Entomological Society. 8, 16-20. Hall, G.M., 1992. Fish processing technology. in: Ockerman, H.W. (Ed.), Fishery byproducts. VCH publishers, New York, USA, pp. 155-192. Ichhponani, J.S., Malik, N.S., 1971. Evaluation of de‐oiled silkworm pupae meal and corn‐steep fluid as protein sources in chick rations. British Poultry Science. 12, 231-234. International Monetary Fund, 2010. International Monetary Fund Primary Commodity Prices.
23
ACCEPTED MANUSCRIPT Justi, K.C., Hayashi, C., Visentainer, J.V., de Souza, N.E., Matsushita, M., 2003. The influence of feed supply time on the fatty acid profile of Nile tilapia
PT
(Oreochromis niloticus) fed on a diet enriched with n-3 fatty acids. Food
RI
Chemistry. 80, 489-493.
Katayama, N., Ishikawa, Y., Takaoki, M., Yamashita, M., Nakayama, S., Kiguchi, K.,
SC
Kok, R., Wada, H., Mitsuhashi, J., 2008. Entomophagy: A key to space
NU
agriculture. Advances in Space Research. 41, 701-705. Keller, S., 1990. Making profits out of seafood wastes. Proceedings of the International
MA
Conference on Fish By-Products Anchorage. University of Alaska Sea Grant, Alaska, 248 pp.
D
Kitsa, K., 1989. Contribution des insectes comestibles à l'amélioration de la ration
AC CE P
29, 511–519.
TE
alimentaire au Kasaï-Occidental. Zaïre-Afrique: économie, culture, vie sociale.
Kroeckel, S., Harjes, A.G.E., Roth, I., Katz, H., Wuertz, S., Susenbeth, A., Schulz, C., 2012. When a turbot catches a fly: Evaluation of a pre-pupae meal of the Black Soldier Fly (Hermetia illucens) as fish meal substitute — Growth performance and chitin degradation in juvenile turbot (Psetta maxima). Aquaculture. 364– 365, 345-352.
Lepage, G., Roy, C.C., 1984. Improved recovery of fatty acid through direct transesterification without prior extraction or purification. The Journal of Lipid Research. 25, 1391–1396. Mile, R.D., Chapman, F.A., 2006. The benefits of fish meal in aquaculture diets. The fish site.
24
ACCEPTED MANUSCRIPT Newton, G.L., Booram, C.V., Barker, R.W., Hale, O.M., 1977. Dried Hermetia illucens larvae meal as a supplement for swine. Journal of Animal Science. 44, 395-400.
PT
Nguyen, T.N., Davis, D.A., Saoud, I.P., 2009. Evaluation alternative protein sources to
of the World Aquaculture Society. 40, 113-121.
RI
replace fish meal in practical diets for juvenile tilapia, Oreochromis spp. Journal
SC
Ogunji, J.O., 2004. Alternative protein sources in diets for farmed tilapia. CAB
NU
International Publishing (Oxford, UK). Nutrition Abstracts and Reviews. Series B, Livestock Feeds and Feeding. 74, 23N-32N.
MA
Ogunji, J.O., Kloas, W., Wirth, M., Schulz, C., Rennert, B., 2006. Housefly Maggot Meal (Magmeal): An Emerging Substitute of Fishmeal in Tilapia Diets,
D
Conference on International Agricultural Research for Development; Deutscher
TE
Tropentag, Bonn, Germany.
AC CE P
Ogunji, J.O., Kloas, W., Wirth, M., Neumann, N., Pietsch, C., 2008. Effect of housefly maggot meal (magmeal) diets on the performance, concentration of plasma glucose, cortisol and blood characteristics of Oreochromis niloticus fingerlings. Journal of Animal Physiology and Animal Nutrition. 92, 511-518. Phelps, R.J., Struthers, J.K., Mayo, J.L., 1975. Investigations into the nutritive value of Macrotermes falciger (Isoptera: Termitidae). Zoologica Africana. 10, 123-132. Ramos-Elorduy, J., 1999. Insects as intermediate biotransformers to obtain proteins. in: Dickinson-Bannack, F., Garcia-Santaella, E. (Eds.), Homo sapiens: An endangered species, towards a global strategy for survival. Proceedings of the 4th World Academic Conference on Human Ecology 1993, Yucatan, Mexico, pp. 157–165.
25
ACCEPTED MANUSCRIPT Ramos-Elorduy, J., 2005. Insects: a hopeful food source. in: Paoletti, M.G. (Ed.), Ecological implications of minilivestock: potential of insects, rodents, frogs and
PT
snails. Science Publishers, New Hampshire, USA, pp. 263-291.
RI
Ramos-Elorduy, J., 2008. Energy supplied by edible insects from mexico and their nutritional and ecological importance. Ecology of Food and Nutrition. 47, 280-
SC
297.
NU
Ramos-Elorduy, J., Conconi, M., 1994. Edible insects in the world (liste des espèces, lieux de consommation et ethnies qui les consomment), Abstracts 4th
MA
International Congress of Ethnobiology, Lucknow, India, pp. 311. Ramos-Elorduy, J., Pino, J.M., Correa, S.C., 1998. Insectos comestibles del Estado de
D
México y determinación de su valor nutritivo. Anales del Instituto de Biología,
TE
Universidad Nacional Autónoma de México. Serie zoología. 69, 65-104.
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Ramos-Elorduy, J., Moreno, J.M.P., Prado, E.E., Perez, M.A., Otero, J.L., de Guevara, O.L., 1997. Nutritional value of edible insects from the state of Oaxaca, Mexico. Journal of Food Composition and Analysis. 10, 142-157. Ramos-Elorduy, J., Medeiros-Costa, E., Ferreira-Santos, J., Pino-Moreno, J.M., Landero-Torres, I., Ángeles-Campos, S.C., García-Pérez, A., 2006. Estudio comparativo del valor nutritivo de varios coleoptera comestibles de México y Pachymerus nucleorum (Fabricius, 1792) (Bruchidae) de Brasil. Interciencia. 31, 512-516. Ramos‐Elorduy, J., 1997. Insects: A sustainable source of food? Ecology of Food and Nutrition. 36, 247-276. Sealey, W.M., Gaylord, T.G., Barrows, F.T., Tomberlin, J.K., McGuire, M.A., Ross, C., St-Hilaire, S., 2011. Sensory Analysis of Rainbow Trout, Oncorhynchus mykiss,
26
ACCEPTED MANUSCRIPT Fed Enriched Black Soldier Fly Prepupae, Hermetia illucens. Journal of the World Aquaculture Society. 42, 34-45.
PT
Sheppard, C., 2002. Black soldier fly and others for value added manure management.
RI
Athens, GA.: University of Georgia. Department of Entomology and Animal Science.
SC
St-Hilaire, S., Sheppard, C., Tomberlin, J.K., Irving, S., Newton, L., McGuire, M.A.,
NU
Mosley, E.E., Hardy, R.W., Sealey, W., 2007. Fly prepupae as a feedstuff for rainbow trout, Oncorhynchus mykiss. Journal of the World Aquaculture Society.
MA
38, 59-67.
Tacon, A.G.J., 1993. Feed ingredients for warm water fish: Fish meal and other
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processed feedstuffs. FAO Fisheries Circular No. 856.
TE
Teguia, A., Beynen, A.C., 2005. Alternative feedstuffs for broilers in Cameroon.
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Livestock Research for Rural Development. 17, Art. #34 Online. Teotia, J.S., Miller, B.F., 1974. Nutritive content of house fly pupae and manure residue. British Poultry Science. 15, 177-182. Venegas-Venegas, E., Rincón-Cervera, M.A., Guil-Guerrero, J.L., 2011. Triarachidonoyl glycerol purification process. J Am Oil Chem Soc. 88, 323-328. Ward, J.H., 1963. Hierarchical grouping to optimize an objective function. Journal: Journal of The American Statistical Association. 58, 236-244. Yhoung‐Aree, J., Puwastien, P., Attig, G.A., 1997. Edible insects in Thailand: An unconventional protein source? Ecology of Food and Nutrition. 36, 133-149.
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ACCEPTED MANUSCRIPT Figure legends
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Figura 1. Cluster analysis of amino acids similarity among insect species, fish meal and
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soybean meal
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acids (using fish meal and soybean reference)
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Figure 2. Differences between insect orders studied in the percentage of essential amino
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fish meal and soybean reference)
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Figura 3. Differences between insect orders studied in the percentage of fat acids (using
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soybean meal
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Figure 4. Cluster analysis of fatty acids similarity among insect species, fish meal and
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Table 1. Order, stage of developement and origin of the species of insect analised
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Table 2. Proximate analysis (% dry matter) of selected insects, fish meal and soybean meal
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Fish meal Soybean meal
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CP % 65.7 ± 1.3 34.6 ± 0.3 58.4 ± 0.4 53.5 ± 0.4 48.3 ± 0.9 61.8 ± 0.3 46.8 ± 1.1 40.9 ± 0.9 36.2 ± 0.3 40.7 ± 0.4 53.5 ± 4.4 59.0 ± 1.5 46.9 ± 4.1 40.1 ± 0.4 46.3 ± 0.6 56.0 ± 2.0 73.1 ± 3.3 66.0 ± 5.0 64.9 ± 0.5 74.4 ± 1.0 58.5 ± 0.5 73.0 ± 0.8 50.4 ± 0.2
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EE % 15.9 ± 1.4 11.8 ± 1.5 30.1 ± 0.7 38.0 ± 0.3 20.1 ± 0.7 27.0 ± 3.2 16.5 ± 0.0 5.8 ± 0.6 18.0 ± 1.6 15.6 ± 0.1 28.4 ± 1.5 26.6 ± 1.0 31.3 ± 1.6 33.7 ± 0.7 28.3 ± 0.6 23.6 ± 0.3 15.9 ± 0.2 17.6 ± 0.2 23.2 ± 0.6 8.8 ± 0.0 29.9 ± 0.5 8.2 ± 0.0 3.0 ± 0.0
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ASH % 7.8 ± 0.2 6.6 ± 0.6 3.5 ± 0.2 2.5 ± 0.3 8.0 ± 0.1 7.2 ± 0.1 6.1 ± 0.1 13.9 ± 0.4 9.3 ± 0.3 19.7 ± 0.1 4.9 ± 0.9 4.9 ± 0.2 6.5 ± 1.5 8.4 ± 2.9 3.9 ± 0.1 8.8 ± 0.1 5.6 ± 0.0 3.7 ± 0.1 4.8 ± 0.1 5.1 ± 0.1 4.0 ± 0.0 18.0 ± 0.2 7.8 ± 0.0
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PeA C RfL C TmL C ZmL C CvL D CmL D CmP D EtL D HiL D HiP D LsL D LsP D MdL D MdP D PtL D PtP D AdA O AaA O GaA O HlA O LmA O FM SM
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Specie Phyllognathus excavatus Rhynchophorus ferrugineus Tenebrio mollitor Zophoba morio Calliphora vicina Chrysomya megacephala Chrysomya megacephala Eristalis tenax Hermetia illucens Hermetia illucens Lucilia sericata Lucilia sericata Musca domestica Musca domestica Protophormia terraenovae Protophormia terraenovae Acheta domestica Anacridium aegyptium Gryllus assimilis Heteracris littoralis Locusta migratoria
NFE % 10.6 ± 0.1 47.0 ± 1.3 8.0 ± 0.2 6.0 ± 1.1 23.6 ± 0.1 4.0 ± 3.4 30.6 ± 1.1 39.4 ± 1.1 36.5 ± 1.0 24.0 ± 0.7 13.2 ± 4.6 9.5 ± 0.1 15.3 ± 4.0 17.8 ± 0.3 21.5 ± 0.1 11.6 ± 2.2 5.4 ± 0.3 12.7 ± 4.8 7.0 ± 0.3 11.7 ± 1.0 7.6 ± 0.1 0.8 ± 0.7 38.8 ± 0.3
Values are means ± SD of triplicate determinations. EE - Crude fat. CP - Crude protein. NFE -
Nitrogen-free extract
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Table 3. Amino acid (as a percentage of total amino acids) of selected insects, fish meal and soybean meal
FM SM
PHE 4.17 5.81 4.29 5.00 7.12 7.17 6.73 6.64 6.88 6.22 7.42 7.02 7.01 6.86 8.10 7.15 4.23 5.00 4.10 3.63 3.84 5.38 5.79
PRO 5.80 7.38 7.17 5.62 4.71 4.56 4.76 5.06 6.16 5.56 4.95 4.83 5.33 5.37 4.91 4.98 5.84 7.21 6.20 6.75 7.46 4.76 4.99
THR 4.10 4.00 4.49 4.33 4.86 4.51 5.02 5.02 5.39 4.95 5.38 4.60 4.87 5.28 4.78 4.83 4.10 4.49 4.11 3.90 4.28 6.26 4.17
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MET 1.42 0.45 0.64 0.76 2.16 2.22 2.76 2.37 1.50 3.26 3.36 3.08 3.00 3.44 2.30 2.55 1.49 2.36 1.10 1.02 0.54 2.93 1.01
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LYS 6.34 6.18 6.03 5.82 7.99 8.53 7.87 8.45 7.60 7.31 7.66 7.91 8.36 7.57 8.23 7.89 6.16 5.73 6.46 6.01 6.33 8.78 6.34
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LEU 7.59 6.63 8.65 8.25 6.69 6.89 6.96 7.62 6.87 6.83 6.43 6.96 6.75 6.57 6.29 6.77 8.69 7.28 8.06 8.88 8.31 7.81 8.01
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ILE 5.71 5.85 5.87 6.36 5.09 4.85 5.23 6.16 5.76 5.34 5.05 5.10 4.89 5.20 5.20 5.08 5.31 5.16 5.05 5.34 5.27 5.04 5.47
MA
HIS 4.19 3.62 3.64 3.87 5.02 5.08 5.20 4.22 5.29 5.16 5.12 5.18 4.68 5.17 5.48 5.35 2.93 4.04 3.03 2.69 2.98 7.86 3.28
D
ARG 5.76 4.82 6.14 5.72 8.57 7.83 8.99 7.69 8.24 8.05 8.84 7.67 6.83 8.76 7.49 7.71 8.53 9.63 9.23 7.93 7.58 7.42 8.03
TE
PeA C RfL C TmL C ZmL C CvL D CmL D CmP D EtL D HiL D HiP D LsL D LsP D MdL D MdP D PtL D PtP D AdA O AaA O GaA O HlA O LmA O
AC CE P
Specie
TYR 4.26 9.84 4.18 6.28 6.36 6.68 6.02 5.21 6.35 7.14 6.64 6.29 5.79 5.91 7.43 7.10 4.91 5.75 4.31 4.51 4.48 3.91 2.93
VAL 7.15 6.69 7.61 7.55 5.93 5.77 6.14 6.52 6.31 6.34 5.88 6.03 6.08 6.08 5.99 6.05 6.99 6.64 6.60 7.48 7.01 5.56 5.45
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Table 4. Fatty acid content (as a percentage of total fatty acid) of selected insects, fish meal and soybean meal
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18:3 n3
18:4 n3
20:5 n3
22:5 n3
22:6 n3
PeA C
0.9 ± 0.1
0
0
0
0
Other 1.5 ± 2.1
28.7 ± 3.0
Satura.
Monoun.
Ω -6
Ω-3
Polyuns.
58.1 ± 0.5
11.8 ± 0.4
0
11.8 ± 0.4
RI
Specie
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Table 4. Fatty acid content (as a percentage of total fatty acid) of selected insects, fish meal and soybean meal (cont.)
1.9 ± 0.1
0
0
0
0
0.1 ± 0.0
42.5 ± 0.4
44.4 ± 0.0
13.0 ± 0.4
0
13.0 ± 0.4
1.1 ± 0.0
0
0
0
0
0.6 ± 0.0
22.2 ± 0.1
45.8 ± 0.1
31.5 ± 0.1
0
31.5 ± 0.1
ZmL C CvL D
1.4 ± 0.0
0
0
0
0
1.6 ± 0.1
38.8 ± 0.2
35.7 ± 0.4
24.0 ± 0.0
0
24.0 ± 0.0
0.2 ± 0.3
1.3 ± 0.1
1.4 ± 0.0
0.7 ± 0.1
0.4 ± 0.0
2.9 ± 0.0
28.5 ± 0.2
40.7 ± 0.4
24.1 ± 0.1
3.9 ± 0.3
28.0 ± 0.1
CmL D
0.6 ± 0.0
0.4 ± 0.0
1.3 ± 0.1
0.6 ± 0.1
0
2.3 ± 0.6
35.9 ± 1.2
30.6 ± 0.1
28.5 ± 0.5
2.9 ± 0.4
31.3 ± 0.7
CmP D
0.5 ± 0.0
0.7 ± 0.1
1.5 ± 0.1
0.4 ± 0.0
0.2 ± 0.2
3.0 ± 1.1
35.4 ± 0.6
35.4 ± 0.7
23.0 ± 0.3
3.2 ± 0.4
26.2 ± 0.1
6.5 ± 1.2
41.7 ± 1.4
50.3 ± 0.2
1.6 ± 0.0
0
1.6 ± 0.0
0.1 ± 0.2
67.1 ± 0.6
16.9 ± 0.2
15.2 ± 0.4
0.7 ± 0.1
15.9 ± 0.6
0.6 ± 0.0
65.8 ± 0.1
32.6 ± 0.1
1.1 ± 0.0
0
1.1 ± 0.0
0
0
0
0
0.7 ± 0.1
0
0
0
0
HiP D
0
0
0
0
0
LsL D
0.4 ± 0.0
0.2 ± 0.0
0.1 ± 0.0
0
LsP D
0.3 ± 0.0
0
0.4 ± 0.0
0
MdL D
0.3 ± 0.0
0.1 ± 0.0
0.1 ± 0.1
0
MdP D
0.3 ± 0.0
0.2 ± 0.0
0.1 ± 0.1
0
PtL D
0.8 ± 0.0
1.1 ± 0.1
0.8 ± 0.0
0
PtP D
0
0.9 ± 0.0
1.8 ± 0.0
0
0
AaA O
17.9 ± 0.4
0
0
GaA O
1.8 ± 0.0
0
0.7 ± 0.1
HlA O
19.4 ± 0.1
LmA O
4.7 ± 0.1
FM
0.2 ± 0.3
SM
6.9 ± 0.2
7.8 ± 0.3
27.8 ± 0.1
55.0 ± 0.1
8.8 ± 0.4
0.7 ± 0.0
9.5 ± 0.4
6.0 ± 0.0
28.8 ± 0.4
54.3 ± 0.2
10.3 ± 0.1
0.7 ± 0.0
11.0 ± 0.1
0
7.2 ± 0.0
32.6 ± 0.1
52.7 ± 0.2
7.1 ± 0.0
0.5 ± 0.1
7.6 ± 0.1
0
8.5 ± 0.1
30.0 ± 1.1
54.1 ± 0.8
7.0 ± 0.4
0.6 ± 0.1
7.5 ± 0.4
0
6.3 ± 0.1
27.1 ± 0.2
44.8 ± 0.1
19.2 ± 0.1
2.7 ± 0.1
21.9 ± 0.2
0
8.3 ± 1.9
26.6 ± 1.6
43.5 ± 0.1
19.2 ± 0.2
2.5 ± 0.0
21.7 ± 0.2
0
0
1.3 ± 0.1
34.2 ± 0.1
21.3 ± 0.1
43.2 ± 0.1
0
43.2 ± 0.1
0
0
1.0 ± 0.0
30.3 ± 1.7
38.7 ± 1.0
30.0 ± 0.7
0
30.0 ± 0.7
0
1.0 ± 0.7
34.0 ± 0.6
27.5 ± 0.4
36.9 ± 0.2
0.7 ± 0.1
37.5 ± 0.3
D
0.8 ± 0.0
0
0
0
0
0.4 ± 0.0
0
0
2.7 ± 0.3
27.7 ± 0.6
27.6 ± 0.3
41.7 ± 0.1
0.4 ± 0.0
42.1 ± 0.1
0
0
0
0
0.1 ± 0.1
36.4 ± 0.1
47.8 ± 0.5
15.9 ± 0.4
0
15.9 ± 0.4
1.9 ± 0.0
14.1 ± 0.2
2.7 ± 0.1
16.1 ± 0.1
6.0 ± 0.4
36.1 ± 1.1
20.6 ± 0.7
2.7 ± 0.2
34.7 ± 0.2
37.3 ± 0.0
0
0
0
0
5.6 ± 0.8
24.0 ± 1.9
15.1 ± 0.3
55.4 ± 0.8
0
55.4 ± 0.8
AC CE P
TE
0.8 ± 0.0
AdA O
NU
0
HiL D
MA
EtL D
SC
RfL C TmL C
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Figura 1.
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Figure 2.
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Figura 3.
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Figure 4.
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Highlights The aim of this paper is to show the insects as a sustainable alternative source to fish meal and soy meal We have evaluated nutritionally (protein, lipid, aminoacids, fatty acids) several insects. We have classified the insects regarding the similarity to the fish meal and soy meal
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