Livestock Research for Rural Development 22 (3) 2010 Notes to Authors LRRD Newsletter

Citation of this paper

Potential use of larval diet disposal from Medfly mass-rearing as alternative livestock feed

T Mastrangelo, J Silva*, A L Abdalla*, M R Peçanha* and J M Melges Walder

Food Irradiation and Radioentomology Laboratory, CENA/USP, Piracicaba, SP, Brazil
* Animal Nutrition Laboratory, CENA/USP, Piracicaba, SP, Brazil


Several Sterile Insect Technique facilities for national programs against fruit flies are being constructed around the world. Along with the mass-rearing of millions of sterile fruit flies per week, large amounts of larval diet are wasted every day and environmental problems could be diminished with its re-utilization as alternative feedstuff for small ruminants. The objective of this work was to characterize the nutritional composition of the larval diet used for the Medfly reared in Brazil and to evaluate its in vitro degradability, also comparing the results to others tropical by-products.


There was no significant decrease in any of the nutritional constituents between the initial and wasted diet due to Medfly larval feeding. The diet presented mean levels of DM, CP, EE, NDF, ADF, CF, ash and OM of 925, 175, 24, 337, 210, 89, 40 and 967 g/kg DM respectively. Phenolic and tannin contents were very low and the larval diet could be safe for the ruminal digestion. In vitro fermentation was higher during the first 6-8 h post-incubation and the linear curves for cumulative gas production for either initial and wasted diets did not differ. Therefore, the larval diet disposal from the mass-rearing of Medfly has potential as an alternative livestock feed.  

Keywords: alternative feedstuff, by-product, fruit fly


Fruit flies (Diptera: Tephritidae) are considered some of the most damaging plant pests in the world, due to their wide range of hosts, direct damages to fruit growers and by the fact that quarantine regulations against them affect global trade (Malavasi and Zucchi 2000). A particular specie, the Mediterranean fruit fly or Medfly (Ceratitis capitata Wiedemann), still causes annual losses of billions of dollars in most tropical and subtropical areas of the world (IAEA 2005, 2008).


A component of area-wide integrated pest management against Medfly is the Sterile Insect Technique (SIT), which consists in the releasing of sterile insects to introduce sterility into wild populations (Knipling 1955). Following its original development over 50 years ago, SIT has been used for suppression and/or eradication by many countries, e.g. Medfly was eradicated from North America and most of Central America, Chile and regions from South Africa. As the demand for alternative environmental friendly methods of pest control continuous increasing, additional SIT facilities are being constructed around the world (IAEA 2008).


The Center for Nuclear Energy in Agriculture (CENA/USP), supported by FAO/IAEA, has made researches on Medfly since the 1970’s. On 2006, CENA started mass-rearing a new genetic sexing strain, the tsl-Vienna 8 (Caceres 2002), which is being used for population suppression with only sterile male releases at the San Francisco River Valley and other fruit-growing areas from Brazil Northeastern, through a partnership with Biofactory MOSCAMED Brasil.


One of the SIT steps is the mass-rearing of the insect (Parker 2005). Insect adults and larvae must be fed in laboratory with artificial diets in order to maintain colonies and guarantee a weekly supply of millions of sterile males (Cohen 2004). For the Medfly strain cited above, larvae take five to six days to grow and start jumping of the diet to pupate on sand or vermiculite (Walder 2002). After this, the larval diet is discarded. At the El Piño biofactory in Guatemala, a key component of the world’s longest running Medfly Eradication and Barrier Program, more than 2 billion sterile males per week and 31 tons of larval diet per day are produced (Teixeira, pers. commun.). Biofactory MOSCAMED Brazil intends to produce initially 200 million sterile males per week and the amount of wasted diet will also reach a high scale.


The Brazilian biofactory is located in an irrigated agricultural pole of the Middle-Lower San Francisco River Valley, one of the poorest areas of the country. A study conducted by the Brazilian Agricultural Research Corporation (EMBRAPA) about the commercialization of sheep and goat meat at the municipalities of Petrolina/Juazeiro, located beside the San Francisco River, revealed that this activity was estimated in US$ 4.5 million per year (Moreira et al 1998), showing the importance of the small ruminant production for the regional economy.


The ruminant livestock productivity in that semi-arid region is very constrained by several factors as the shortage of good quality feed and poor sanitary conditions. According to FAO, Brazilian production of ovine meat is still stocked in 76 000 tons per year since 2004 (ANUALPEC 2008). A potent tool to improve production, especially for those growers who do not have access or cannot afford conventional feed resources, is the supplemental feeding with agro-industrial by-products (Blache et al 2008).


The Brazilian agribusiness generates more than 300 million tons per year of agro-industrial by-products (Teruya 1999). There are many successful examples of alternative feedstuffs throughout the world, such as willow cuttings, lupin seed, feed blocks, wasted dates, brewer’s grain, citrus pulp, cotton seed meal, sugarcane bagasse and alternative pasture species. These materials can present advantages as attractive nutritional composition, high availability and low cost. Their incorporation in ruminant feeding represents a rational end for such residues and contributes for the establishment of sustainable animal production systems (Blache et al 2008; Cabral Filho 1999).


Publications about the waste disposal of fruit flies mass-rearing facilities or its re-utilization as feeding for livestock are very rare (Parker 2005). The rational use of Medfly larval diet as an alternative feed resource requires the previous knowledge of its nutritional characteristics and digestible parameters.


Therefore, the objective of the present study was to characterize the nutritional composition of the larval diet for Medfly reared at CENA and to evaluate its in vitro digestibility, aiming its future use as supplement in ruminant feeding. Results were also compared to other tropical by-products.      


Material and methods 

The larval diet for mass-rearing of Medfly was obtained at the Food Irradiation and Radioentomology Laboratory of CENA. The ingredients of the diet are wheat germ (6%), wheat flour (6.5%), brewer yeast (9.5%), sugar (12%), sodium benzoate (0.3%), citric acid (1.7%), tetracycline (0.02%), distillated water (55%) and dried sugarcane bagasse (9%). To prepare the diet, the liquid components and the tetracycline are mixed first and then the solid ingredients are added. Chemical analysis and digestibility were made at the Animal Nutrition Laboratory (LANA) of CENA.     


Diet samples and chemical analysis


Chemical composition of the larval diet was determined before and after Medly larvae feeding in order to observe any diet changes due to larval development. Three random samples (0.5 kg) were collected on the first day (moment of seedling Medfly eggs over the diet) and on the last day of larval cohort (moment when the diet is ready to be discarded but few larvae remain inside). After collection, samples were frozen to kill the remaining larvae.


Samples were assayed to dry matter (DM), crude protein (CD), ether extract (EE), ash and organic matter (difference between DM and ash) according to the Association of Official Analytical Chemists (AOAC 1995). Crude fiber (CF), acid detergent fiber (ADF) and neutral detergent fiber (NDF) were analyzed in DM according to Van Soest and Wine (1967).


The larval diet was also analyzed for total phenols, total tannins and condensed tannins. Total phenols (TF) and total tannins (TT) were determined with the Folin-Ciocalteau reagent. A calibration curve was prepared using tannic acid (Merk GmbH, Darmstadt, Germany). TF and TT were calculated as tannic acid equivalents and expressed as eq-g/kg DM. Condensed tannins (CT) were measured by the HCl-butanol method and the colorimetric data were converted to leucocyanidin equivalents and expressed as eq-g/kg DM (FAO/IAEA 2000; Makkar 2000; Porter et al 1986).      


In vitro gas production measurement


Adult sheep (Santa Ines species) fitted with rumen cannula and maintained on a basal diet of Brachiaria decummbens pasture plus a concentrate of maize gain, soybean meal and mineral salt were used as rumen content donors. Rumen digesta were collected before the morning feeding. Cumulative gas production profiles were generated using methodology of Theodorou et al (1994) as modified by Mauricio et al (1999) with pressure transducer and data logger (PDL 200, LANA/CENA-USP, Piracicaba/SP, Brazil). To the samples (0.5 g), it was added 25 mL of the inoculums plus 50 mL of buffer, and gas pressure measurements were made at 0, 6, 14 and 24 h post-inoculation. The amount of gas was estimated according to Bueno et al (2005).  


Statistical analysis


To study the chemical composition, the one-way analysis of variance F-Test was calculated for the means of DM, CP, EE, CF, NDF, ADF, ashes and OM at the 5% level of significance and the Student’s t-test (α = 0.05) was used to compare means. Analyses were performed by the statistical program SAS 9.1. (PROC GLM, SAS Institute 2003). Linear regression for the cumulative gas production was performed according to Sokal and Rohlf (1981) and the accumulated production in 24 h was compared by the Student’s t-test (α = 0.05).


Results and discussion  

Chemical composition


In general, wide variations exist in the chemical composition of investigated feedstuffs around the world. Nutrient contents of the larval diet are presented in Table 1.

Table 1. Chemical composition of the diets, before and after the larval development


Larval Diet

t-Tests (LSD)

Initial Diet

Wasted Diet

(g/kg of FM )


40 ºC

394 ± 10.3a ab

366 ± 35.7 a

(F = 0.56; P = 0.49)


924 ± 3.9 a

926 ± 1.5 a

(F = 0.22; P = 0.69)

(g/kg of DM)


167 ± 8.7 a

182 ± 7.9 a

(F = 1.73; P = 0.26)


18 ± 4.8 a

29 ± 4.1 a

(F = 2.95; P = 0.16)


318 ±67.3 a

355 ± 61.3 a

(F = 0.16; P = 0.71)


181 ± 45.6 a

237 ± 47.6 a

(F = 0.74; P = 0.44)


76 ± 30.4 a

100 ± 3.5 a

(F = 0.28; P = 0.63)


35 ± 5.1 a

43 ± 7.3 a

(F = 0.76; P = 0.43)


969 ± 1.9 a

963 ± 3.4 a

(F = 2.21; P = 0.27)

a     Standard error.

b     Original means in this table. Means within lines followed by the same letter are not significantly different, ANOVA at P ≤ 0.05.

There is no significant decrease in any of the nutritional constituents between the initial and wasted diet due to Medfly larval feeding. The diet showed DM values that are close to those found in tropical fruit processing by-products as pineapple (Ananas comosus), acerola (Malpighia glabra), guava (Psidium guajava), passion fruit (Passiflora edulis) and melon (Cucumis sp.) (322-621.1 g/kg) (Lousada Junior et al 2006). Levels of DM at 100°C were relatively high, ranging from 924.1±3.9 to 926.1±1.5 g/kg.


The larval diet has CP contents that are similar to sorghum grain (105 g/kg), soybean husk (119 g/kg), corn germ (93 g/kg), rice meal (137 g/kg) and tropical fruit processing by-products (83.5-173.3 g/kg) (Lousada Junior et al 2005, 2006). Silva (1995) presented CP values of cotton seed meal between 400 and 500 g/kg. Santana and Souza (1984) found low concentrations (around 25 g/kg) for crude sugarcane bagasse. Machado (2001) classified soybean and cotton seed meals as suitable protein sources (CP > 200 g/kg) and citrus pulp as an energetic source (CP < 200 and CF < 180 g/kg). Leucaena leucocephala leafs have CP contents about 193-240 g/kg (Godoy 2007), but its phenolic and tannin levels act as anti-nutritional factors. The CP content ranges from 47.7 for coconut meal to 495.6  g/kg for soybean meal (Chumpawadee et al 2007).


Although not significant, there is a CP increment between the initial and wasted diet, which might have occurred due to the presence of Medfly larvae and uric acid produced during their growth. Therefore, comparing to other alternative livestock feeds, in regards to DM and CP values, the wasted larval diet could satisfy the minimum requirement for a healthy ruminal gear.


The crude fat fraction or EE content was also higher in the wasted diet (29.2 ± 4.1 g/kg), probably also caused by the presence of larvae. Insects can store large amounts of lipids in the fat body, since these reserves are important e.g. to support metamorphosis (Pontes at al 2008). According to Nestel et al (2003), there is a high accumulation of lipids before the pupariation and the fat body of larvae can represent almost 9% of the total body weight in Medfly.   


These EE levels are not so far from the ones of some native pasture leafs (around 53 g/kg) (Palmquist and Jenkins 1980). Lousada Junior et al (2006) reported EE contents of tropical fruit by-products ranging from 11.9 to 60 g/kg. The EE level is relevant for various ruminants because the DM intake trends to decrease with higher fat concentrations (Berchielli et al 2006). According to Rodrigues et al (2003), cashew nut meal (DM = 932.7 g/kg; CP = 221.5 g/kg; EE = 359.7 g/kg; CF = 62.4 g/kg; ash = 30.9 g/kg) can be included up to 24% in diets for sheep raised in feedlot.


Wide variations were observed for CF, NDF and ADF (Table 1), probably caused by the sugarcane bagasse origin which is part of the larval diet. During the year, materials of different ages and sugarcane varieties are collected from several producers. In general, crude sugarcane bagasse presents high contents of fiber (450 g/kg) and lignin (230 g/kg), having low digestibility in cattle (Santana and Souza 1984). According to Mertens (1998) and Lousada Junior et al (2006), some by-products present NDF values of 523 g/kg (brewer’s grain), 386 g/kg (alcohol distillery by-product), 185 g/kg (soybean meal), 114 g/kg (corn grain), 213 g/kg (citrus pulp) and 562-735 g/kg (tropical fruit by-products). Cândido et al (2004) stated that the addition of dried passion fruit by-product up the level of 14% by the time of Elephant grass (Pennisetum purpureum) ensiling promotes reductions on its NDF and hemicelluloses levels, sustaining adequate EE contents. The ADF for citrus and tomato pulp and apple pomace were estimated in 210-220, 366-435 and 252-302 g/kg respectively (Bampidis  and Robinson 2006; Mirzaei-Aghsaghali and Maheri-Sis 2008). Chumpawadee et al (2007) found ADF levels ranging from 94.8 g/kg (soybean meal) to 456.5 g/kg (Leucaena sp. meal).


No significant difference was found to ash and OM contents from initial and wasted diets, being around 40 and 965 g/kg respectively. MM values ranges from 34.3 to 145.7 g/kg for tropical fruit by-products (Lousada Junior et al 2005, 2006). Wadhwa et al (2006) studying non-conventional vegetable wastes (e.g. cabbage leaves, cauliflower leaves and pea pods and vines) found OM values ranging from 85 to 172 g/kg.


Mahgoub et al (2008) estimated the ash content of a commercial by-products based concentrate in 118 g/kg. Ash levels in Cassava sp. leaves from Africa is in the range 550-161 g/kg (Asaolu 1998). Chumpawadee et al (2007) working with various feed mills and organizations in the North East of Thailand, reported that ash contents ranged from 10.8 for coconut meal to 176.9 g/kg for Leucaeana sp. meal. As for most of the tropical forages, the larval diet could not attend exclusively the mineral demand of ruminant animals (Berchielli et al 2006).


Differences between initial and wasted diet were observed for TF, TT and CT (Table 2). However, their values were very low and, assuming that the CT threshold considered safe for ruminal digestion is up to 30-40 eq-g leucocyanidin/kg DM (Barry et al 1986), the larval should offer no risk for small ruminants.

Table 2.  Phenolic and tannin contends of the diets, before and after the larval development

Constituent (eq-g TA or leucocyanidin /kg DM)

Initial Diet

Wasted Diet

t-Tests (LSD)

Total phenols

4.4 ± 0.15a bb

7.8 ± 0.21 a

(F = 182.3; P = 0.005)

Total tannins

2.2 ± 0.21 b

3.8 ± 0.11 a

(F = 48.2; P = 0.02)

Condensed tannins

0.4 ± 0.01 a

0.2 ± 0.02 b

(F = 780; P < 10-3)

a Standard error; b Original means in this table. Means within lines followed by the same letter are not significantly different, ANOVA at P ≤ 0.05.

Much work has been done on tanniniferous browses in several countries as some levels of tannins can limit the utilization of many fodder trees and shrubs in tropical regions (Balogun et al 1998; Kumar and Vaithiyanathan 1990). Vitti et al (2005) determined TP, TT and CT contents for tropical legumes, e.g. Leucaena leucocephala (25.7, 21.6 eq-g TA/kg and 12.7 eq-g leucocyanidin/kg), Sesbania sesban (28.7, 24.3 eq-g TA/kg and 23.5 eq-g leucocyanidin/kg) and Cajanus cajan (8.2, 5 eq-g TA/kg and 0.5 eq-g leucocyanidin/kg). 


Godoy (2007) reported similar CT contents for Calopogonio sp. (0.4 eq-g leucocyanidin/kg) and Medicago sativa (0.2 eq-g leucocyanidin/kg). Cabral Filho et al (2005) obtained higher apparent DM degradability for sorghum grains with lower CT levels and correlations between CT content and gas production following polyethylene glycol (PEG) addition. Guimaraes-Beelen et al (2006), studying forages from a semi-arid Brazilian area, found CT values higher than 100 g/kg for all phonological stages of Mimosa hostiles, M. caesalpinifolia and Bauhinia cheilantha.


Studies had shown that the consumption of plants presenting a CT concentration of 30-40 g/kg was associated to positive effects on digestion and that protein digestibility is not affected at tannin concentrations lower than 20 g/kg DM (Barry and McNabb 1999; Poncet and Rémond 2002). However, Vitti et al (2005) cast doubt on the generalization that small amounts of condensed tannins (20–40 g/kg) produce beneficial effects or that high levels (>50 g/kg) are necessarily harmful, proposing that it is not yet possible to predict beneficial or harmful nutritional effects from total tannin concentrations per se. That is why in vivo trials are essential to obtain precise information about voluntary intake, digestibility, nitrogen retention and weight gain.


 In vitro gas production


Figures 1 and 2 depict the gas production over a 24 h period.

Figure 1.  In vitro gas production on time for diets before and after larval development

In vitro fermentation was higher during the first 6-8 h either for initial and wasted diet. It might have occurred due to the high amount of sugar of the larval diet. Sugar is used as phagostimulant for Medfly larvae and represents 12% of the diet content. Although the accumulated gas production for the two diets did not differ significantly (F = 3.3; P = 0.12), gas production was slighter lower in the wasted diet, what was probably caused by the consumption of sugar and other nutrients by larvae.


The slopes of the linear regression curves (Figure 2) did not differ (F = 0.19; P = 0.67). Biologically, parallelism of curves can be interpreted to mean that changes in activity per unit change in rate are the same (i.e., potency as defined by Finney (1971)).

Figure 2.  Linear regression of cumulative gas production on time for diets before and after larval development

According to Machado (2001), the cumulative gas production in 24 h for sugarcane bagasse, rice husk, Leucaena leucocephala, brewer’s grain, and cotton seed meal was < 50 mL/g DM, whilst it was > 100 mL/g DM for citrus pulp (with a potential production of 372.3 mL/g DM). The same author states that feedstuffs with high contents in fiber, e.g. sugarcane bagasse and Leucaena leucocephala, can present long lag times.                                       





This study was made possible by a scholarship from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).



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Received 26 November 2009; Accepted 10 January 2010; Published 1 March 2010

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