Welfare of the Mealworm (Tenebrio molitor) Breeding With Regard to Nutrition Value and Food Safety

Po a ina s o Slo ak Jou nal o Food Sciences Volume 11 460 No. 1/2017 Po a ina s o Slo ak Jou nal o Food Sciences ol. 11, 2017, no. 1, p. 460-465 doi: h ps://dx.doi.o g/10.5219/779 Recei ed: 12 Ma ch 2017. Accep ed: 19 June 2017. A ailable online: 14 July 2017 a www.po a ina s o.com © 2017 Po a ina s o Slo ak Jou nal o Food Sciences, License: CC BY 3.0 ISSN 1337-0960 (online) WELFARE OF THE MEALWORM ( TENEBRIO MOLITOR ) BREEDING WITH REGARD TO NUTRITION VALUE AND FOOD SAFETY Anna Adámko á, Ma in Adámek, Jiří Mlček, Ma ie Bo ko co á, Ma ina Bednářo á, Lenka Kouřimská, Jose Skácel, E a Ví o á ABSTRACT Li es ock wel a e is an impo an condi ion o ob aining high-quali y and sa e ood. Acco ding o he legisla ion edible insec s a e classi ied as li es ock; and o his eason i is necessa y o comply wi h he edible insec wel a e condi ions. This a icle ocuses on selec ed wel a e condi ions o mealwo m (Teneb io moli o ) b eeding, wi h special ocus on he a con en in luenced by di e en b eeding empe a u e (17 °C, 23 °C and 28 °C). Maximum a con en 24.56% was obse ed a 23 °C. To ob ain maximum a con en his appea s o be he op imal b eeding empe a u e. Ano he e alua ed aspec was he nu i ional s ess and a way o killing, and hei impac on a con en , which showed o dec ease wi h he nu ien s ess. The mos decline was de ec ed owa ds he end o he obse a ion pe iod. The analysis showed ha in e ms o p ese a ion o he a con en , he bes way is killing by eezing, due o he me abolism slowdown. We also analysed he con en o hea y me als in a mealwo m la ae using cyclic ol amme y wi h subsequen e alua ion. In he measu ed sample concen a ions o hea y me als did no exceed he maximum allowable concen a ion o hea y me als in his commodi y. F om his poin o iew mealwo m appea s o be a sa e ood. Keywo ds: wel a e; Teneb io moli o ; nu i ional s ess; way o killing; hea y me al INTRODUCTION Animal wel a e is a condi ion, when he animal o ganism ies o deal wi h he su ounding (B oom, 1986). Wel a e is de ined as a s a e o ul ilmen o all ma e ial and imma e ial condi ions, which a e p e equisi es o he heal h o he animal, and a e also in acco dance wi h i s en i onmen (Angelo ičo á and Polačko á, 2015; Doležal, Bílek and Dolejš, 2004). Webs e (2016) desc ibed he i e eedoms o animal wel a e. I is eedom om hunge , hi s , discom o , pain, inju y, disease, ea and dis ess, and he exp ession o no mal beha iou ( an Huis e al., 2013). Acco ding o he Regula ion (EC) No. 1069/2009 insec b eeding is conside ed as b eeding o common li es ock. Because o his i is necessa y o ollow he ules o insec wel a e as well as wi h o he li es ock. Tha ’s why Eisemann e al. (1984) sugges ed, ha insec should be gi en he bene i o he doub ( an Huis e al., 2013). Among he species wi h he g ea es po en ial as ood and eed wi hin he Eu opean Union a e mealwo m (Teneb io moli o ), gian mealwo m (Zophobas a a us), lesse mealwo m (Alphi obus diape inus), g ea e wax mo h (Galle ia mellonella), silkwo m (Bombyx mo i), house c icke (Ache a domes icus) and A ican mig a o y locus (Locus a mig a o a mig a o ioides) (EFSA, 2015). Mealwo m (Teneb io moli o ) in gene al belongs o he ood pes s (Sch oeckens ein, Meie -Da is and Bush, 1990; Wang, Liao and Chen, 2012), howe e , hanks o i s nu i ional alues, easy b eeding and minimal en i onmen al bu den i has a po en ial o become a ully- ledged sou ce o p o ein and a o mankind (Wang, Liao and Chen, 2012). Nu i ionally, i is impo an in e ms o p o ein and a , and he e is a p esump ion o good use in he ood indus y (Pa k e al., 2014). Ano he mealwo m bene i could be biodeg ada ion o o ganic was e o p o eins (Veldkamp e al., 2012). Based on he abo e ad an ages, possibili ies o ea ing his species in space a e being explo ed (Ka ayama e al., 2008). F om he economical poin o iew, mealwo m b eeding is cheap and as (Wang, Liao and Chen, 2012). Acco ding o Li, Zhao and Liu (2013) i s li e cycle is sho , egg s age las s 3 – 9 days, la al s age 26 – 76 days and pupal s age 5 – 17 days. Rea ing usually akes place in b eeding anks wi h eed a he bo om (o en ce eal meal, milk powde and bone meal) (Hů ka, 2005). G ow h a e and size o indi iduals is a ec ed by se e al ac o s, including empe a u e, humidi y, ligh in ensi y, composi ion o he eed and densi y o specimens wi hin he b eeding anks (Wu e al., 2009). One o he main ac o s in luencing he g ow h speed is he empe a u e (Xu e al., 2012). Wi h g owing Po a ina s o Slo ak Jou nal o Food Sciences Volume 11 461 No. 1/2017 empe a u e he li e cycle sho ens. In 20 °C he leng h o he li e cycle was 97 days, and in 32 °C only 60 days (Xu e al, 2012). Howe e , wi h g owing empe a u e he weigh o pupae and adul specimens ends o dec ease om ce ain empe a u e le el (Kim e al., 2015). Ano he ac o is he insec eed. Sui able eed is necessa y o main aining good heal h and op imal specimen pe o mance (Dussu ou e al., 2016). Howe e , eely li ing animals do no necessa ily ha e he access o he eed ha e lec s hei nu i i e needs. The in luence o he nu i ional composi ion o eed on he nu i ional alue o insec s had been in es iga ed (Li, Zhao and Liu, 2013). Nu i i e s ess o small b eeding space leads o cannibalism (Wu e al., 2009) and dec eases he b eeding yield. Killing is ca ied ou in di e en ways, mos commonly by eezing, di ec g inding, o cooling and subsequen killing by boiling. Me hods o killing insec s, howe e , should be chosen so as o a oid insec su e ing ( an Huis, 2013). P io o he slaugh e , dead indi iduals mus be so ed ou . Due o inadequa e sa e y e iew o edible insec s as a no el ood o humans in he EU, i is necessa y no only o know he ules o b eeding and p o ide su icien wel a e, bu also o know he e ec o ea ing insec s o human heal h and sa e y isks (EFSA, 2015). EFSA (2015) d aws a en ion o he lack o sa e y e iews o he insec consump ion and a ious isks, including he con en s o hea y me als (mainly Cd, Pb, Hg, and As) and calls o se ing limi s o sa e consump ion. This a icle ocuses on he impac o nu ien s ess as a wel a e aspec , and ways o killing on he nu i ional alue ( a con en ) o he inal p oduc . Fu he mo e, he a icle deals wi h he sa e y o edible insec s in e ms o he con en o hea y me als (Cd and Pb), which ha e a signi ican impac on human heal h. MATERIAL AND METHODOLOGY Ma e ial Insec La ae o mealwo m (Teneb io moli o ) we e used o he analysis (Figu e 1). Samples we e bough in he pe supplies s o e K mi a Hos i ice. Feed Whea b ans wi h he ollowing nu i ional alues we e used as eed ( alues o 100 g o p oduc ): Ene gy: 1210 kJ / 292kcal, Fa 5.3 g, o which sa u a ed a y acids 0.88 g, ca bohyd a es 24.9 g, o which suga s 2.2 g, ib e 40.2 g a p o ein 16.2 g, sal 0.1 g. Company: Coun y Li e, s. .o., Be oun 1. Chemicals  Pe ol e he 40/65, p.a., CAS No.: [8032-32-4], Ing. Pe Š ec – PENTA s. .o., P aha,  HNO3 65%, p.a., CAS No.: [7697-37-2], Ing. Pe Š ec – PENTA s. .o., P aha,  H2O2 30%, p. a., CAS No.: [7722-84-1], Ing. Pe Š ec – PENTA s. .o., P aha,  CdCl2 p. a. – M . 183,32, CAS No.: [10108-64-2], Fluka analy ical, Sigma Ald ich,  PbCl2 p. a. – M . 278,11, Lachema, n. p., B no, CZ,  Deionized wa e – 18.2 MOhm cm, Milli-Q, Millipo e. All chemicals we e o analy ical eagen g ade o equi alen analy ical pu i y. Me hods Insec p ocessing In he i s expe imen , mealwo m la ae we e di ided in o h ee expe imen al g oups. These expe imen al g oups we e placed in a plas ic boxes in a he mos a (HS 62A, Chi ana) by ea ing empe a u e o 17 °C, 23 °C and 28 °C. The g oups we e ed ad libi um. Be o e analysis, la ae in he las and penul ima e ins a de elopmen ( he ull leng h o he body jus be o e pupa ion) we e collec ed om b eeding. Subsequen ly he la ae we e le o s a e o 24 hou s and hen killed wi h boiling wa e (100 °C). The p epa ed samples we e d ied a 105 °C, homogenized and s o ed in a cold s o age box a 4 – 7 °C un il analysis. Fo he second expe imen , wo g oups o mealwo ms we e b ed in oom empe a u e 22 °C ±2 °C and di ided by he nu ien s ess. The i s g oup was ed whea b ans ad libi um du ing he whole es . The o he g oup was in nu i ional s ess, which means wi hou eed o 27 and 39 days. Be o e analysis, la ae in he las and penul ima e Figu e 1 La ae o mealwo m (Teneb io moli o ) Po a ina s o Slo ak Jou nal o Food Sciences Volume 11 462 No. 1/2017 ins a de elopmen ( he ull leng h o he body jus be o e pupa ion) we e collec ed om b eeding. Subsequen ly he la ae we e le o s a e o 24 hou s. Each g oup was hen di ided in o wo sub-g oups. The i s sub-g oup was killed by eezing (-18 °C) in he eeze and he second one was killed by boiling wa e (100 °C). Dead la ae we e d ied a 105 °C, homogenized and s o ed in a cold s o age box a 4 – 7 °C un il analysis. Fa con en de e mina ion The a con en de e mina ion was pe o med by ex ac ion using Soxhle me hod (Soxhle , 1879) on he Ge ha d Sox he m SOX414 (C. Ge ha d GmbH & Co. KG, Ge many). App oxima ely 5 g o d ied and homogenized samples (wi h he accu acy o 0.0001 g) we e pu in o ex ac ion himbles and ex ac ed wi h 150 ml o pe oleum e he ia cold wa e ex ac ion (p og am: 70 °C o 120 minu es). The ex ac ion lask was hen d ied a 103 °C and weighed un il a cons an sample weigh was a ained. Hea y me als con en 0.1 g o homogenized sample was pu in o a ial, and hen 2 mL HNO3 wi h 65% concen a ion was added. Me als we e being ex ac ed o 24 hou s a oom empe a u e and hen hea ed o each he empe a u e 110 °C o 1 hou . Subsequen ly, 200 µL 30% H2O2 was added and he sample was kep wa m o ano he 30 minu es. A e cooling he sample was dilu ed 5 imes wi h deionized wa e ( / ) (18.2 MOhm cm, Milli-Q, Millipo e). P io o he analysis he sample was dilu ed 10 imes using an ace a e bu e ( / ). De ec ion o selec ed me als (Cd, Pb) was ca ied ou using cyclic ol amme y wi hin he ange U = <-1000; +400 >mV wi h scanning speed 10 mV s-1. Accumula ion ime was cond = 45 s wi h po en ial Econd = -1000 mV. To e alua e me als, analy ic elec ochemical s a ion µAUTOLAB, Type III/ FRA 2 was used wi h NOVA 1.11 applica ion. Ag/AgCl Re e ence Elec ode Me ohm AG, Swi ze land was used as a e e ence elec ode. P Rod Elec ode Me ohm AG, Swi ze land, was used as auxilia y elec ode. Wo king elec ode was c ea ed using he hick laye me hod on a ce amic subs a e (Al2O3). Senso size was 25.4 mm ×7.25 mm × 625 μm. The sil e duc ou come was c ea ed by ESL 9562-G pas e (ESL Elec oscience, England). Wo king elec ode wi h 3 mm diame e was c ea ed using DuPon 7102 pas e (DuPon , USA). ESL 4917 pas e (ESL Elec oscience, England) was used o c ea e he p o ec i e co e laye . Measu ed cha ac e is ics we e de i ed and analysed in Excel 2013 (Mic oso Co po a ion, USA). S a is ical analysis Da a ob ained om chemical analysis was p ocessed using MS Excel (Mic oso Co po a ion, USA), o calcula e basic pe cen age o a con en . These da a was hen p ocessed using he S a is ica 12 (S a So , Inc., USA) applica ion. Figu e 2 Fa con en in mealwo m (Teneb io moli o ) -dependency on he b eeding empe a u e (boxplo ). Table 1 Fa con en in mealwo m (Teneb io moli o ) and i s dependency on he b eeding empe a u e. Sample Fa (%) TM 17 °C 14.67 TM 23 °C 24.56 TM 28 °C 23.32 M M ± SE M ± SD 17 °C 23 °C 28 °C Tempe a u e 12 14 16 18 20 22 24 26 28 30 Fa (%) Po a ina s o Slo ak Jou nal o Food Sciences Volume 11 463 No. 1/2017 RESULTS AND DISCUSSION B eeding empe a u e is one o he mos impo an ac o s a ec ing he de elopmen speed (Xu e al., 2012). In ou s udy, we examined he a con en dependency on he empe a u e (17 °C, 23 °C and 28 °C). Table 1 and Figu e 2 show ha he a con en a 17 °C is lowe han a 23 °C, bu u he inc ease o he empe a u e o 28 °C did no lead o g ea e change. These a e, howe e , he ini ial esul s, which will be u he e i ied. While di e en s udies show he gene al in luence o empe a u e on he edible insec nu i ional alue, a ailable sou ces do no s a e any speci ic dependence o nu i ional alues on he b eeding empe a u e (Oonincx and an de Poel, 2011). Xu e al. (2012) in es iga ed he op imum empe a u e o b eeding mealwo m (Teneb io moli o ) and compa ed i e di e en empe a u es (20 °C, 23 °C, 26 °C, 29 °C a 32 °C). Thei esul s showed ha he mealwo m de elopmen ge s as e wi h g owing empe a u e. Also he numbe s o laid eggs g ew (max. a 29 °C) (Xu e al., 2012), howe e , he weigh o pupa and adul dec eased (Kim e al., 2015). The change in weigh and size o specimens in di e en empe a u es can sugges he change o hei nu i i e alue. Role o nu i ion is o p o ide he needed nu ien s in o he body ia he eed, in o de o p ese e li e, g ow h, ep oduc ion, good heal h and pe o mance. The body can elie e he lack o ood (nu i ional s ess) by d awing ene gy om adipose issue, mine al sa u a ion om skele ons, ni ogenous subs ances sa u a ion o mo e economical exc e ion o ce ain elemen s. The deple ion o ce ain nu ien wi hou e illing may esul in dea h o he indi idual. Lack o a s educes he iabili y, slows g ow h, educes ep oduc ion, pe o mance and leads o diseases; long- e m de iciency has le hal e ec s (Ho niako á e al., 2010). To obse e he e ec o nu i ional s ess, insec was di ided in o wo expe imen al g oups. The i s one was ed ad libi um; he o he was in nu i ional s ess by s a ing o 27 and 39 days. In he second g oup he cannibalism occu ed mo e o en, as also men ioned by E ens e al. (2012). Subsequen ly, bo h g oups o s a ing insec s we e di ided in o sub-g oup killed by eezing (-18 °C) and boiling wa e (100 °C). The esul s o he a con en show ha wi h he leng h o s a ing he a con en dec eases (see Figu e 3). A di e ence was de ec ed be ween he g oups o bo h ways o killing. This end became mo e signi ican owa ds he end o he expe imen . I ’s supposed ha he eason is dec easing o he a deposi s in insec bodies due o hei consumma ion by basal me abolism (Na ion, 2015). Changes in a con en in insec bodies as well as insec beha iou we e obse ed also by (Dussu ou , 2016) in an s. Dec ease o he a con en was de ec ed a e educing he nu ien concen a ion in he die , when he a io o p o eins o ca bohyd a es was inc eased, and when he an s we e s a ing. Reg ession model shows ha he a con en was de e mined almos exclusi ely by he ca bohyd a e in ake. The e was also a change in he a con en among a ious cas es (Dussu ou , 2016). Howe e , nu i ional s ess can be caused no only by he lack o eed as a whole, bu also by he lack o a speci ic nu ien . Fo example Simpson a Raubenheime (2012) s a e ha excessi e consump ion o ca bohyd a es leads o obesi y when eed is low in p o ein wi h a io o ca bohyd a es, while he lack o ca bohyd a es in he die leads o a lack o ene gy. Figu e 3 Mealwo m (Teneb io moli o ) a con en dependency on he leng h o s a a ion and he way o killing. 0 5 10 15 20 25 30 0 5 10 15 20 25 30 35 40 Fa con en (g.100 g-1) Lengh o s a a ion (days) F eezing Boiling Po a ina s o Slo ak Jou nal o Food Sciences Volume 11 464 No. 1/2017 Some ways o killing insec , desc ibed by E ens e al. (2012), a e conside ed humane, bu as he s a es, some p ocesso s ejec hem o inancial easons. Bene i o eezing, o example, is ha by pu ing insec in o he eeze , i s me abolism slows down (hibe na ion), un il i eezes. F eezing should be ca ied ou quickly o p e en issue damage (D dák, 1989). Conside ing he nu i ional s ess, he e was no di e ence be ween he ways o killing. Howe e , Figu e 3 shows ha he g oup killed by eezing had a highe a con en han he g oup killed by boiling wa e . We suppose ha he loss o a in he second g oup may be caused by a ex ac ion in o he boiling wa e . When using he a o p o ec ion om he cold and eeze, he losses we e smalle han wi h boiling. Hea y me al concen a ion in ood and eed o animal b eeding is an impo an opic o heal h sa e y and has an impac on he consume s’ heal h (Cho anco á e al., 2014; Lukáčo á e al., 2014). In ou esea ch we ocused on he de ec ion o cadmium and lead in edible insec using ca bon elec ode c ea ed wi h he echnology o hick laye s. A e c ea ing he calib a ion cu es he cadmium and lead concen a ion was e alua ed in mealwo m la ae (Teneb io moli o ) in las and penul ima e ins a de elopmen , b ed in 23 °C and ed by whea b ans ad libi um. Bo h cadmium and lead concen a ion was below he limi o de ec abili y o he used senso (de ec ion limi is 2 µg.L-1 o lead and 3 µg.L-1 o cadmium). The senso can be used o de ec he le els o cadmium and lead, s a ed as a maximum limi in Regula ion (EC) No. 1881/2006 (Pb 0.5 mg.kg-1 ha is 2.41 µg.L-1 and Cd 4.46 µg.L-1). Al hough hese a e ini ial esul s, gained wi h using senso wi h de ec ion limi s sligh ly below he maximum allowable concen a ion in his commodi y, he con en o cadmium and lead seems o be sa e o using insec as human ood. Poma (2017) e alua ed nine mine al elemen s in his s udy. In mealwo m (Teneb io moli o ) he con en he de ec ed was: Cd 0.06 mg.kg-1 and con en o Pb was lowe han he de ec ion limi . The le el o hea y me als e alua ed in his s udy was lowe han eques ed by he Regula ion (EC) No. 1881/2006 and lowe o compa able wi h o he commodi ies o animal o igin. CONCLUSION Ou wo k was ocused on he impac o wel a e on he nu i ional alue and sa e y o he mealwo m (Teneb io moli o ), as a po en ial ood sou ce. As an analysis o a con en indica es, b eeding empe a u e – as one o he wel a e ac o s – signi ican ly a ec s he a con en . O he e alua ed wel a e ac o was nu i ional s ess, while he expe imen al g oup loaded wi h nu i ional s ess had lowe a con en . I can be concluded, ha nu i ional dep i a ion a ec s no only he animal wel a e, bu also nu i ional yield wi h a u he impac on he economic aspec s o b eeding. Fo comme cially aised li es ock he momen o dea h is an impo an pa o animal wel a e, ha ing an impac on he nu i ional alue and quali y o he mea ob ained. The way o killing also in luenced he nu i ional alues o mealwo m la ae in ou s udy. F om he iewpoin o a yield, killing by eezing can be ecommended. Al hough he ques ion o hea y me als con en is no pa o wel a e, i is an impo an aspec o he ood sa e y. Based on ou s udy, we can conclude ha hea y me al con en o he analysed species was lowe han he maximum pe mi ed le els o hea y me als in ood. F om his poin o iew, he consump ion o mealwo m la ae is sa e. REFERENCES Angelo ičo á, M., Polačko á, D. 2015. Assessmen o wel a e and egg p oduc ion o laying hens Mo a ia SSL in small-scale b eeding. Po a ina s o, ol. 9, no. 1, p. 365- 374. h ps://doi.o g/10.5219/514 B oom, D. M., 1986. 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Con ac add ess: Anna Adámko á, Czech Uni e si y o Li e Sciences P ague, Facul y o Ag obiology, Food and Na u al Resou ces, Depa men o Quali y o Ag icul u al P oduc s, Kamýcká 129, 165 21 P ague 6 – Suchdol, Czech Republic, E-mail: [email p o ec ed] Ma in Adámek, B no Uni e si y o Technology, Facul y o Elec ical Enginee ing and Communica ion, Depa men o Mic oelec onics, Technická 3058/10, 616 00 B no, Czech Republic, E-mail: [email p o ec ed] Jiří Mlček, Tomas Ba a Uni e si y in Zlin, Facul y o Technology, Depa men o Food Analysis and Chemis y, Va ecko a 275, 760 01 Zlin, Czech Republic, E-mail: [email p o ec ed] Ma ie Bo ko co á, Mendel Uni e si y in B no, Facul y o Ag onomy, Depa men o Zoology, Fishe ies, Hyd obiology and Ag icul u e, Zemědělská 1, 613 00 B no, Czech Republic, E-mail: ma ie.bo ko co [email protected] Ma ina Bednářo á, Mendel Uni e si y in B no, Depa men o In o ma ion Technology, Zemědělská 1, 613 00 B no, Czech Republic, E-mail: bedna o [email protected] Lenka Kouřimská, Czech Uni e si y o Li e Sciences P ague, Facul y o Ag obiology, Food and Na u al Resou ces, Depa men o Quali y o Ag icul u al P oduc s, Depa men o Mic obiology, Nu i ion and Die e ics, Kamýcká 129, 165 21 P ague 6 – Suchdol, Czech Republic, E-mail: [email p o ec ed] Jose Skácel, B no Uni e si y o Technology, Facul y o Elec ical Enginee ing and Communica ion, Depa men o Mic oelec onics, Technická 3058/10, 616 00 B no, Czech Republic, E-mail: [email p o ec ed] E a Ví o á, B no Uni e si y o Technology, Facul y o Chemis y, MRC-labo a o y o bio echnology and bioma e ials esea ch, Pu kyňo a 118, 612 00 B no, Czech Republic, E-mail: [email p o ec ed]