International Journal of Applied Science and Technology Vol. 5, No. 6; December 2015

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Interntionl Journl of Applied Science nd Technology Vol. 5, No. 6; December 2015 Assessment of the implementtion Degree in Hndling Prctices Which Contribute to Reducing Fungl Incidence nd its Consequent Afltoxins Production in Penut Kernels Sofi G. Cuggino Fcultd de Ciencis Agropecuris Universidd Ncionl de Córdob Alejndr Pérez Agostini Fcultd de Ciencis Agropecuris Universidd Ncionl de Córdob Abstrct The penut kernel intended for direct humn consumption, while this hs high nutritionl vlue, is very likely to be ltered t different production stges. Thereby, fungl presence in grins decreses its qulity s food nd cn produce mycotoxins, cusing tht grins be unfit for consumption. The purpose of this study ws to evlute the implementtion degree in those hndling prctices which contribute to reducing fungl incidence nd its consequent fltoxins production in penut kernels. Five representtive smples of penut kernels were evluted. All these of direct consumption t three res the centrl zone from Córdob Province (Argentin). Vribles evluted were: fungl incidence, fltoxins content, nd implementtion percentge in hndling prctices relted to fltoxins contmintion. Determintions were crried out through vrince nlysis model, with which res s fixed effects bout sid model were compred to detect significnt differences; between mens, ANOVA ws performed; nd for comprisons, Tukey test (p<0.05); t lst, correltions were clculted. Fungl incidence levels remined not directly relted to fltoxins concentrtion. The higher implementtion percentge the GAP ws ssocited with lower fltoxins concentrtion, even if environmentl conditions were predisposing. Keywords: Penut - Afltoxins- Fungl incidence - Good Agriculturl Prctices. Introduction The penut (Archis hypoge L.) ws produced nd mrketed primrily s rw mteril the oil industry nd for direct humn consumption (Blengino, 2014). The Argentine penut gribusiness sector ws locted minly from Córdob Province nd constitutes very importnt regionl economy dedicted lmost exclusively for export, given tht 95% the produce ws intended for interntionl mrkets (Bols de Cereles de Córdob, 2012). The penut is high nutritionl vlue food for direct humn consumption, which needs sustinble production system to ensure its sfety. Therefore, penut kernel is considered very likely to be ltered t different production stges, due to biotic nd biotic fctors. In prticulr, fungl presence in grins decreses its qulity s food: cuses discolortion, odors, nutritionl nd chemicl disorders; nd cn produce mycotoxins, which bring bout tht penuts or their derivtives be unfit for consumption (Christensen, 1982; Mrch & Mrinelli, 2005; Pster & Bullermn, 1988; Schneider & Sieber, 1999; Pitt et l., 2012). The fungl gener Aspergillus, Penicillium, Rhizopus nd Fusrium re frequently detected in penut kernels (Mores & Mriotto, 1985; Cvllo et l., 1994; nd Cvllo et l., 2005). Among gener mentioned, Aspergillus stnds out s the most importnt fltoxins producer (Chulze, 2005; Kumed & Asso, 2001; Mrch & Mrinelli, 2005). Azizeh et l. (1990), Vn Egmont et l. (2004), nd Cornejo & Villrroel (2007) hve estblished tht low levels of fungl coloniztion hve lso presented lower levels of fltoxins. Accordingly, the A. flvus detection could be n indictor mycotoxins presence, becoming simple method to prevent toxins contmintion. While the mycotoxins presence ws trditionlly problem ssocited with storge, reserch hs shown tht contmintion cn occur before hrvest (Mrch & Mrinelli, 2005). 143

ISSN 2221-0997 (Print), 2221-1004 (Online) Center for Promoting Ides, USA www.ijstnet.com Best wy to prevent fltoxins emergence in grins is dopt preventive mesures for fungl controlling t ll production stges (WHO, 2012). For this, it is necessry to identify, mong crop hndling prctices, those which fvor toxigenic fungl prolifertion nd its consequent mycotoxins genertion. About, Chulze (2005) rgues tht hndling strtegies to minimize or eliminte fltoxins contmintion begins t countryside nd ends t industril process. Thus, Good Agriculturl Prctices (GAP) nd Good Mnufcturing Prctices (GMP) implementtion decreses fltoxins contmintion risks in penut kernels. (Bongiovnni et l., 2012). So fr, hs not been estblished t Argentine production zone the implementtion degree in those prctices which contribute to reducing fungl incidence nd its consequent fltoxins production. The purpose of this study ws to evlute the implementtion degree in those hndling prctices which contribute to reducing fungl incidence nd its consequent fltoxins production in penut kernels, from centrl zone the Córdob Province (Argentin). Mterils & Method This reserch ws crried out with penut kernels intended for direct humn consumption t three res of production the penut centrl zone from Córdob Province (Argentin). The res were identified s: Are 1 (Onctivo), Are 2 (Psco), nd Are 3 (L Plestin). Five smples per re t Cmpign 2011-2012 were worked. Smples of 8.8 lb weight were tken representtively, ccording to Rule XIII (SAGPyA, 1994). The picking ws performed with Geis-Cl invert torn mchine, consisting of swth from four grooves. Completed the period t swth, its top ws removed with Aipridec mchine of single row. The pods were plced into polyethylene plstic burlp bgs nd were sent immeditely to lbortory for nlysis. Then, these were husked mnully nd evlutions were conducted with penut kernels of confectionery ctegory. Vribles evluted were: Fungl incidence: It ws determined through modified blotter test method (Mthur & Kongsdl, 2003). Superficilly, penut kernels with helthy ppernce were disinfected with sodium hypochlorite solution t 1.5% for two minutes nd were rinsed twice with sterile distilled wter. Lter, these were plced on sterile filter pper moistened with sterile distilled wter on plstic trys (6.7 x 9 x 1.6 in) formerly disinfected with lcohol t 95%. Finlly, 3 replictes of 50 grins per ech smple evluted were plced to incubte into chmber t 69.8 ± 35.6 ºF under UV light during cycles 12 hours light & 12 hours drk. Assessments in individul grins were performed seven dys fter sowing, through binoculr lens nd stereoscopic microscope (40x). Fungi were clssified by techniques commonly used in mycology nd with help of keys (Mthur & Kongsdl, 2003). Vribles registered were: percentge the totl fungl incidence nd infected grins with A. flvus, A. niger, Alternri, Penicillium, Fusrium nd Rhizopus (Bringel et l., 2001). Totl fltoxins: It ws preceded through direct competitive ELISA technique for totl fltoxins by Romer Lbs AgrQunt. For extrction, it ws worked with 4.4 lb of penut with husk for ech smple. Rndomly spced, 1.76 lb of penut ws husked, which then were milled with Romer Series II so tht 75% pssed through No. 20 mesh. 0.04 lb of milled smple were weighed, nd it ws derived to lcoholic extrction with solution methnol/wter t 70/30 (v/v). Concentrtions were clculted by extrpolting opticl density (OD) with respective clibrtion curve obtined. Results were expressed in prts per billion (ppb). Evlution the implementtion degree in hndling prctices, relted to fltoxins contmintion: Dt collection ws performed using semi-structured personl surveys to technicl mngers for ech re, under GAP for penut crop (Bongiovnni et l., 2012; Pedelini, 2012). The checklist ws grouped ccording to ech stge the production process, from sowing to trnsport goods to plnt. Assessment of ech item ws estblished s impct the executed tsk in reltion to occurrence or possible fungl contmintion. From ddition of vlues obtined in ech item, totl score ws obtined; nd from this, implementtion percentge ws clculted. Determintions were crried out with vrince nlysis model, with which res considered s fixed effects bout sid model were compred. Men vlues of ech vrible were subjected to sttisticl nlysis to detect significnt differences through ANOVA; comprisons were mde with Tukey test (p<0.05) nd correltions were clculted using InfoStt sttisticl progrm (Di Rienzo et l., 2014). Results & Discussion Averge vlues the totl fungl incidence obtined showed sttisticlly significnt differences t Are 2 reltive to other res evluted (Figure 1). 144

Interntionl Journl of Applied Science nd Technology Vol. 5, No. 6; December 2015 100 Versión Estudintil 90 Versión Estudintil Versión Estudintil Versión Estudintil Versión Estudintil Versió Versión Estudintil 80 Versión Estudintil Versión Estudintil Versión Estudintil Versión Estudintil Versió tudintil 70 Versión Estudintil Versión Estudintil Versión Estudintil Versión Estudintil Versión Estudin Fungl incidence (%) tudintil 60 Versión Estudintil Versión Estudintil Versión Estudintil Versión Estudintil Versión Estudin 50 b Versión Estudintil 40 Versión Estudintil Versión Estudintil Versión Estudintil Versión Estudintil Versió 30 tudintil 20 Versión Estudintil Versión Estudintil Versión Estudintil Versión Estudintil Versión Estudin tudintil 10 Versión Estudintil Versión Estudintil Versión Estudintil Versión Estudintil Versión Estudin Versión Estudintil 0 Versión Estudintil Versión Estudintil Versión Estudintil Versión Estudintil Versió Are 1 Are 2 Are 3 Figure 1: Totl fungl incidence t three representtive res the penut centrl zone from Córdob Province. Equl letters indicte no significnt differences (Tukey p<0.05). In Figure 2, percentges the infected grins with ech genus t res evluted re observed. Dt indicted tht fungl incidence with different gener wsn't homogeneous between res evluted. Gener detected were: Aspergillus, Penicillium, Rhizopus nd Fusrium; stnding, t Are 2, Penicillium presence which reched vlues of up to 35%. In ll btches, there ws fungl contmintion; nd ll them, these ws lso coincidence with pollutnts mostly found in penut kernels (Cvllo et l., 1994; Cvllo et l., 2005; Mores & Mriotto, 1985; Mzzni, 1989). e b Fungl Incidence (%) d f h f h d h b c b c b c g Are Figure 2: Fungl incidence per genus in penut kernels t three representtive res the penut centrl zone from Córdob Province. Different letters indicte significnt differences between evluted res (Tukey p<0.05). A. flvus incidence between res evluted showed no significnt differences (p<0.05) Figure 2. This fungl specie, potentil precursor the mycotoxins, ws found eqully t three res. So tht, this sitution represents potentil risk which depends on environmentl conditions nd hndling prctices executed, ccording to proposls by Fonsec (1991), Melouk & Shokes (1995) nd Schpovloff et l. (2010). 145

ISSN 2221-0997 (Print), 2221-1004 (Online) Center for Promoting Ides, USA www.ijstnet.com Determintion the totl fltoxins showed significnt differences between res (Figure 3). The Are 2 ws it lowers fltoxins contmintion. 2,0 Versión Estudintil 1,8 n tudintil 1,6 Versión Est 1,4 Totl Afltoxins (ppb) b Versión Estudintil 1,2 n tudintil 1,0 Versión Est 0,8 Versión Estudintil 0,6 n tudintil 0,4 Versión Est 0,2 Versión Estudintil 0,0 n tudintil Versión Estudintil Versión Are Estudintil 1 Versión Estudintil Are Versión 2 Estudintil Versión Are Estudintil 3 Versión Estudint Figure 3: Afltoxins concentrtion in penut smples t three representtive res the penut centrl zone from Córdob Province. Different letters indicte significnt differences (Tukey p<0.05). Results presented by PAHO (1983); Mrch & Mrinelli, (2005); Fernández et l. (2006); nd Cornejo & Villrroel (2007) indicte tht A. flvus presence is closely ssocited with fltoxins formtion. Results found in this reserch didn't show direct correltion between A. flvus percentge nd fltoxins vlues detected (R 2 = 0.03). By compring studied res, different fltoxins concentrtion ws estblished for sme level A. flvus contmintion. Regrding hndling prctices tht reduce fungl incidence nd subsequent fltoxin contmintion, the higher implementtion percentge, ccording to surveyed dt, ws obtined for Are 2 (Tble 1). Tble 1: Implementtion percentge the prctices t three res of production from Córdob Province. Etp Cumplimiento (%) Áre 1 Áre 2 Áre 3 Previo l siembr 25.0 78.3 53.3 Siembr 85.7 85.7 85.7 Crecimiento del cultivo 100.0 100.0 100.0 Cosech 20.0 33.3 20.0 Post cosech 0.0 0.0 0.0 Totl 32.5 52.1 40.5 The comprtive nlysis between evluted res demonstrtes tht, while A. flvus incidence levels showed no significnt differences (Figure 2), the higher implementtion percentge the hndling prctices t Are 2 determined lower fltoxins level (p<0.05). These results re contrsted to proposls by Cornejo & Villrroel (2007) nd Nyirhkizimn et l. (2013), who indicte simple nd direct reltion between percentge the fungl incidence with A. flvus nd fltoxins contmintion level in penut kernels. However, these re consistent with proposls by Chulze (2005) nd Pitt et l. (2012), who hold tht mycotoxin formtion depends not only of fungl incidence, but intercting environmentl fctors such s humidity, temperture nd crop hndling. The predisposing environmentl conditions for fltoxins formtion weren't those tht occurred t pre-hrvest stges t res evluted (tempertures no higher thn 77 ºF, bsence moisture deficit Figure 4). Nevertheless, t post-hrvest stges, during the time of permnence t swth, rinflls ccumulted during the months of April nd My were higher t Are 2 (52 mm), regrding Are 1 (22 mm) nd Are 3 (42 mm). c 146

Interntionl Journl of Applied Science nd Technology Vol. 5, No. 6; December 2015 Figure 4: Rinflls (mm) nd mximum tempertures (ºF) monthly verge for Agriculturl Cmpign 2011-2012 t Ares 1, 2 & 3 (Córdob Grin Exchnge). The higher implementtion percentge the GAP (Tble 1) ws ssocited with lower fltoxins genertion, even if environmentl conditions were predisposing (Are 2). The stges in which this re hd the higher GAP implementtion were: t prior-sowing stge (crop rottion, fllow execution nd mechnicl tillge), t sowing stge (qulity ssurnce seed, determintion sowing time), nd t hrvest stge (decrese permnence t swth). Stges with the lower implementtion percentge the GAP were observed t hrvest stges until the time of entry to processing plnt. Conclusion The fungl incidence levels t studied res remined not directly relted to fltoxins concentrtion, which cnnot be estimted mycotoxin risk just to helth test the penut kernels. The higher implementtion percentge the GAP, relted mycotoxins formtion, ws ssocited with lower fltoxins concentrtion even if environmentl conditions were predisposing. It's noteworthy tht while t different res evluted, the higher contmintion risk ws verified t post-torn stges, it's necessry to implement GAP throughout penut crop cycle, order to encourge grins production of better qulity intended for direct humn consumption. 147

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