Harmful Effects on Oryzaephilus Surinamensis (L.) and Tribolium Castaneum by Food Grade Antioxidants and Their Formulations in Peanut Kernel
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ournal of JFood Chemistry & Nanotechnology https://doi.org/10.17756/jfcn.2017-042 Research Article Open Access Harmful Effects on Oryzaephilus surinamensis (L.) and Tribolium castaneum by Food Grade Antioxidants and their Formulations in Peanut Kernel Daiana Garcia1*, Natalia Soledad Girardi2, María Alejandra Passone1, Andrea Nesci1 and Miriam Etcheverry1 1Research Career, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina 2Doctoral fellow of Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina *Correspondence to: Abstract Daiana Garcia Research Career, Consejo Nacional de The aim of this study was to investigate effect of 2(3)-tert-butyl-4 Investigaciones Científicas y Técnicas hydroxyanisole (BHA) and 2,6-di(tert-butyl)-p-cresol (BHT) and their (CONICET), Argentina formulations on mortality, body weight and total protein content of Oryzaephilus Tel: +54 358-4676113 surinamensis (L.) and Tribolium castaneum, Herbst; two natural pest of stored Fax: +54 358-4676231 E-mail: [email protected] peanut. The effect of free BHA and BHT and their formulations (F-BHA and F-BHT) at dose range to 10 from 45 mM, in peanut kernel was evaluated. BHA Received: April 25, 2017 and BHT free antioxidant showed significant mortality percentages upper than Accepted: June 27, 2017 80%. Besides, microencapsulated antioxidants at 20 and 30 mM significantly Published: July 05, 2017 affected the survival of insect populations after 45 days of exposure. BHA and Citation: Garcia D, Girardi NS, Passone MA, BHT formulation showed insecticidal activity of 100 and 70%, respectively. Nesci A, Etcheverry M. 2017. Harmful Effects Insect weight was significantly affected by all studied treatments and the major on Oryzaephilus surinamensis (L.) and Tribolium castaneum by Food Grade Antioxidants and their effect for O. surinamensis was produced by time especially with the application of Formulations in Peanut Kernel. J Food Chem BHA follow by F-BHA and F-BHT. Body mass weight of T. castaneum showed Nanotechnol 3(3): 86-92. the highest reduction after application of F-BHA, followed F-BHT, BHT and Copyright: © 2017 Garcia et al. This is an Open BHA. Total protein content of insects also was affected by both free antioxidants Access article distributed under the terms of the and formulation treatments. Formulations preserved insecticidal effectiveness Creative Commons Attribution 4.0 International of antioxidants and extended their effect for up to 45 days which could act as License (CC-BY) (http://creativecommons. new strategies to control these insect pests. Besides, sub-lethal levels of these org/licenses/by/4.0/) which permits commercial compounds results in a loss of insects’ weight and change in protein content. use, including reproduction, adaptation, and distribution of the article provided the original Results showed the first step in the toxicity of free and microencapsulated author and source are credited. formulations of BHA and BHT against O. surinamensis and T. castaneum. Published by United Scientific Group Keywords BHA, BHT, Oryzaephilus surinamensis, Tribolium castaneum, Microcapsules Introduction Argentina is one of the major producer and supplier of food worldwide. Córdoba, from the center of this country, is an important peanut producing area. In 2015/16 harvest season Argentina had a total production around of 1.17 million tons ranking seventh among the world’s largest producers of peanut [1]. Primary peanut production is intended for both industrialization and exportation and for the last case there is fluctuation between 0.44 and 0.68 million tons since 2012 [2]. Peanut, in their different presentations, is very little consumed internally which makes all production is oriented to exportation. At the moment industry elaborate peanut for direct consume and flavored peanut, pasta and peanut butter, oil and peanut pellets. Considering that 6% of the raw material can be lost due to fungi contamination and infest with insect during harvest, transport and storage, so it is necessary to have safe methods of preservations and control to the different contaminants. Garcia et al. 86 Harmful Effects on Oryzaephilus surinamensis (L.) and Tribolium castaneum by Food Grade Antioxidants and their Formulations in Peanut Kernel Garcia et al. Two important species of insect pest saw-toothed Preparation of antioxidant formulation grain beetle [Oryzaephilus surinamensis (L.) (Coleoptera; Industrial grade antioxidants, 2(3)-tert-butyl-4 Silvanidae)] and the red flour beetle [Tribolium castaneum hydroxyanisole (BHA) and 2,6-di(tert-butyl)-p-cresol (BHT), Herbst (Coleoptera; Tenebrionidae)] which attack already obtained from Eastman Chemical Company (Kingsport, infested or malformed grains, preferring stored cereal products Tennessee, USA) were used as core material. BHA had a purity and oleaginous seeds [3-5]. Besides, they can acts as vector of 98.5% containing as trace elements sulphated ash 100 μg/g, for potential toxigenic molds such as Aspergillus spp. and citric acid 2.5 μg/g, arsenic 3 μg/g, and heavy metals 10 μg/g. the constant migration of insect populations within granary BHT had a purity of 99% containing as contaminants ash 100 ecosystem efficiently contributes to dispersion of viable fungal μg/g, arsenic 3 μg/g and heavy metals 10 μg/g. Contaminant spores which are carried on the vector’s body surface or are compounds of industrial grade antioxidants did not exceed deposited with its feces [6]. Some research has shown that allowed levels by JECFA [14]. Gelatin (type A, gel strength both type of genera of these insects’ acts as vector of Aspergillus 240 bloom) and gum arabic were used as the wall material. flavus in storage seeds as maize and peanut [7-9]. Control of All other chemicals used in this work were of analytical grade. these types of insects is considered of great importance as they Microcapsules were made by complex coacervation following can remain in continuous contact with raw material in the the methodology proposed by Girardi et al. [13]. Microcapsules storage system, thus they can disperser fungal spores of fungi obtained were frozen at -80 °C during 3 hours and freeze-dried potentially producing mycotoxins. with a chamber (L-T8-A-B3-CT, RIFICOR) pressure <0.05 Some works have demonstrated butylated hydroxyanisole mbar and -45 °C for 72 h. Empty capsules were performed (BHA) and butylated hydroxytoluene (BHT) (phenolic with the same methodology but without the addition of BHA antioxidants) effectiveness against toxigenic molds and these or BHT, in order to be used as control by replacing the core storage pests [10, 11]. However, analysis of antioxidant residual material with peanut oil. levels in stored peanuts showed a fast reduction of these substances, probably due to the environmental and biological Microcosm assay factor interactions [12]. To improve the performance of Insecticidal effect determination: To determine these preservatives “in situ” both antioxidants were subjected insecticidal activity of free chemicals and in their formulations, to a microencapsulation process in order to protect them of 150 g of sterile peanut kernels were distributed into plastic jars different environmental agents according to Girardi et al. [13], of 500 mL capacity and treatments were added at different who reported residual levels after 2 and 3 of storage months. doses: 10, 20 and 30 mM for O. surinamensis and 15, 30, 45 Previous studies showed that both formulations maintained mM for T. castaneum. Fifteen adults of each type of insect were the insecticidal effect against O. surinamensis for more than 45 introduced into a jar and incubate in chamber under controlled days [9]. Insecticidal activity of pure BHA and formulation conditions (25 ± 1 °C, 70 ± 5% r.h.). Assay was done using of both antioxidants were demonstrated in O. surinamensis by three replicates per treatment and number of dead insect was Nesci et al. [7] and Garcia et al. [9], respectively. However, determined at different times during a 45 days holding period. insecticidal activity on T. castaneum and harmful effects on Treatments with empty capsules (without BHA or BHT) and both types of insects it is unknown. To advance in this sense, without capsules were used as control. the proposal of this study was to investigate effect of free and Body weight evaluation: Insects were weight before to microencapsulated BHA and BHT on i) mortality; ii) body start the assay (time 0) and at different period time till the end weight and iii) total protein content in O. surinamensis and T. of the experiment for all conditions (treatment and controls). castaneum. Weight loss was calculated by difference between the data obtained at the different evaluation times. Materials and Methods Protein content evaluation Substrate and insects Protein content extraction: To determine protein content Natural peanuts collected during the harvest season 2014- of insects affected by the presence of free and microencapsulated 2015 from Córdoba, Argentina, with an initial water content antioxidants, 100 g of sterile peanut kernels were distributed into plastic jars of 300 mL capacity with 15 adults of each of 0.67 were used throughout this study. Seeds were sterilized insect species. Antioxidants pure and their formulations were by autoclave at 120 °C for 20 min and water activity (aw) was checked with an AquaLab Water Activity Meter 4TE added at non-lethal doses: 3 and 1 mM of BHA and BHT (Decagon Devices, Inc.) with an accuracy of ± 0.001. Strain respectively, for O. surinamensis while for T. castaneum 25 and of the saw-toothed grain beetle O. surinamensis and red flour 3 mM for BHA and BHT respectively, were used. Jars were beetle T. castaneum were obtained from the Laboratory of incubated in chamber under controlled conditions (25 ± 1 °C, Agricultural Zoology, Faculty of Agronomy, and University 70 ± 5% r.h.). Total proteins were extracted by Lee et al. [15] of Buenos Aires, Argentina. Mixed-sex adults 1-3 weeks old method with some modifications: insects exposed to chemical were used in the assays separately.