Mealworm(Tenebrio molitor) was added to the feed to improve the health of dogs. As the amount of mealworm powder added to dog feed increased from 2.5 to 7.5%, the contents of crude protein, crude fat, crude fiber and crude ash increased. In the old dogs, creatinine and BUN related to nephropathy, triglyceride related to hypertension, calcium related to osteoporosis, globulin related to immunity, and ALT related to hepatic function were in the normal range. Particularly, ALT showed a tendency to increase overall, but the numerical increase tended to decrease as the amount of mealworm powder added. In the young dogs, all investigative items such as BUN and creatine were in the normal range and ALT tended to decrease at 2.5～5.0% treatment of mealworm. As a result, it was shown that adding 2.5～5.0% of mealworm powder to feed effectively improved the hepatic function of old and young dogs.

Insect cuticle tanning (pigmentation and sclerotization) is a complex and vital physiological process that begins with tyrosine and is responsible for production of both melanin- and quinoid-type pigments. In addition, these quinones undergo isomerization to quinone methides and cross-linking reactions with cuticular proteins for cuticle sclerotization. In this study, we studied the functions of TmDDC and TmY-y as well as TmNAT1, TmADC and Tmebony from Tenebrio molitor, which are involved in the tyrosine-derived melanin- and quinoid-type pigment productions, respectively. The temporal and spatial expression patterns of the genes were analyzed by real-time PCR. RNA interference was performed to understand the genetic regulation and molecular mechanism underlying the darkening and hardening of beetle cuticle.

Toll and IMD pathways play an important role in producing antimicrobial peptides (AMPs) through NF-κB in insects. The functions of IκB kinase (IKK) complex regulating the NF-κB signaling cascade have not yet been investigated in Tenebrio model. Here, we identified TmIKK-β (or TmIrd5) which contains 2,112 bp encoding 703 amino acid residues. Domain analysis shows that TmIKK-β contains one Serine/Threonine protein kinases catalytic domain. Developmental expression patterns indicate that TmIKK- β gene was highly expressed in early pupal (P1) and adult (A5) stages. Tissue specific profiles show that TmIKK-β was highly expressed in the integuments in last instar larvae, and fat body and hemocytes in 5 day-old adults. TmIKK-β1 transcripts were strongly induced at 3 and 12 h-post injection of E. coli, and 3 h-post injection of S. aureus or C. albicans in hemocytes. In gut, TmIKK-β transcripts were slightly induced by E. coli (at 6, 9 and 24 h) and C. albicans (at 24 h), while it was not induced by S. aureus challenge. Moreover, it was highly induced at 6 h-post injection of E. coli and then it was gradually decreased in the fat body. To understand the immunological role of TmIKK-β, gene specific RNAi and mortality assay was performed. Depletion of TmIKK-β mRNA leads to increase microbial susceptibility of larvae against E. coli, S. aureus and C. albicans. In addition, induction patterns of fourteen AMP genes in response to microbial challenge was tissue specifically investigated in TmIKK-β–silenced T. molitor larvae. The results suggest that expression of ten AMP genes out of fourteen genes were drastically decreased by TmIKK-β RNAi in fat body, suggesting that TmIKK-β plays an important role in antimicrobial innate immune responses.

It has been well known that IKK-β, -ε and –γ play a pivotal role in IMD pathway. In this study, TmIKK-ε was identified and their functions in countering pathogenic infections were investigated. We identified TmIKK-ε gene which including 2,196 bp nucleotides (encoding 731 amino acid residues). Domain analysis of TmIKK-ε indicates that there is one Serine/Threonine protein kinases catalytic domain. TmIKK-ε gene was highly expressed in 2 day-old pupal stage and the expression was gradually decreased until 1 day-old adults. Then the expression was slightly increased until 4 day-old adult stage. Tissue specific expression of TmIKK-ε mRNA was high in the gut, integuments and hemocytes in last instar larvae, and fat body, Malpighian tubules and testis in 5-daysold adult. In hemocytes, TmIKK-ε was drastically induced by E. coli injection after 3 h and by S. aureus at 3 and 12 h-post injection. In gut, expression level of TmIKK-ε was high at 6 h-post injection of microbial injection. Expression of TmIKK-ε in fat body was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-ε, gene specific RNAi and mortality assay were performed. TmIKK-ε RNAi caused increased larval mortality against E. coli, not S. aureus and C. albicans. Finally, to investigate the induction patterns of Tenebrio fourteen AMP genes in response TmIKK-ε RNAi, three microorganisms were treated into TmIKK-ε-silenced T. molitor larvae. Nine out of fourteen AMP genes were not induced by microbial challenge in TmIKK-β dsRNA-injected group. Taken together, our results indicate that TmIKK-ε may regulates nine antimicrobial peptide genes in response to microbial challenge in T. molitor fat body.

Host defense against pathogen invasion highly relies on immune defense machinery that is controlled by the nuclear factor-κB (NF-κB) of transcription factors. The Toll pathway are well known as an insect innate immune mechanism to protect host itself from invaded pathogens. Basically, in the edible insect, Tenebrio molitor, the Toll pathway is primarily activated by polymeric Lys-type peptidoglycans (PGNs), and components of fungal cell walls, β-1,3-glucan. Based on the current studies, the tremendous study has been focused on recognition and subsequent activation of spätzle in haemolymph, hence, there is a grave gap for intracellular event. Herein, in order to understand intracellular event of Toll signaling pathway, the Dorsal gene were identified. Moreover, domain analyses of TmDorsal2 gene indicate that there are two major domains such as Rel homology domain (RHD), ig-like, plexins, and transcription factors (IPT) domains. Based on the achieved results, TmDorsal2 mRNA was highly expressed in 1-day old pupa. Furthermore, TmDorsal2 was highly expressed in Malpighian tubules and fat body in last instar larvae (LL), and likewise mainly expressed in Malpighian tubules during adult 5-day old period, also the lowest expression of TmDorsal2 was observed in gonads. Moreover, TmDorsal2 mRNA levels after infection with E. coli appreciably went up at 6 and 9h time points. To investigate the effects of TmDorsal2 RNAi on larval susceptibility against various pathogens namely E. coli, S. aureus or C.albicans, dsRNA of TmDorsal2 has been synthesized the larvae dissected after 24h. As a result, TmAttacin1a, 1b and 2, TmDefencine1 and 2, TmTenecin1, 2, 3 and 4, TmCecropin2, TmColeoptericin1 and 2, Thaumatin-like protein 1 and 2 markedly reduced in the gut after injecting all mentioned microbes. In contrast, TmTenecin 2, Thaumatin-like protein 1 and 2 strikingly increased after microbe injection in the fat body. Interestingly, the most AMPs gene expression in whole body experimental case were upregulated. On the horizon, we will investigate effects of TmDorsal1 RNAi on larval susceptibility against various pathogens. Taken together, our studies may aid to understand insect innate immunity.

Tenebrio molitor beetles have been widely used in traditional and folklore medicines to treat various human diseases worldwide. Alzheimer’s disease (AD) is a neurodegenerative disorder and the most prevalent form of dementia in developed and developing countries. The human β-amyloid cleaving enzyme (BACE-1) is a target for AD treatments. This study was performed to determine BACE-1 inhibitory activity of 12 compounds that were extracted from T. molitor adults and their 65 related compounds using a fluorescence resonance energy transfer-based enzyme assay. The results were compared with those of two positive controls for BACE-1, the cell-permeable isophthalamide BACE-1 inhibitor IV and the natural BACE-1 inhibitor, epigallocatechin gallate (EGCG). Based on IC50 values, linoleic acid, ergosterol, palmitoleic acid, and tryptopol (28.41‒46.16 μM) were the most potent BACE-1 inhibitors and the anti-BACE-1 activity of these compounds was lower than either inhibitor IV (13.13 μM) or EGCG (1.12 μM). Multiple regression analysis of the anti-BACE-1 activities of compounds was examined using their IC50 values and the values of the physical parameters (molecular weight (MW), log P, and molecular refraction (MR)) for the 41 fatty acids (R2 = 0.674 (P = 0.044)) and 29 flavonoids (R2 = 0.587 (P = 0.063)). Correlation coefficient (r) analysis showed that MW, log P, and MR may be positively correlated with IC50 of the fatty acids (MW, r = 0.658; log P, r = 0.471; and MR, r = 0.713) and IC50 of the flavonoids (MW, r = 0.547; log P, r = 0.644; and MR, r = 0.591). Further studies will warrant possible applications of T. molitor adults as therapeutic BACE-1 blocker.

Tenebrio molitor beetles have been widely used in traditional and folklore medicines to treat various human diseases worldwide. This study was performed to determine radical scavenging activity of nine compounds that were extracted from T. molitor adults and their 64 related compounds using a 2,2-diphenyl-1-picrylhydrazyl assay. The results were compared with those of three positive controls, ascorbic acid, butylated hydroxytoluene (BHT), and epigallocatechin gallate (EGCG). Based on IC50 values, linolenic acid (77.65 μM) was the most potent antioxidant and the antioxidant activity of this compound was higher than that of either ascorbic acid, BHT or EGCG. The antioxidant activity of protocatechuic acid (IC50, 153.68 μM) and ascorbic acid did not differ significantly. The antioxidant activity of catechin (IC50, 192.32 μM) was higher than that of either BHT or EGCG. Multiple regression analysis of the antioxidant activities of compounds was examined using their IC50 values and the values of the physical parameters (molecular weight (MW), log P, and molecular refraction (MR)) for the 41 fatty acids (R2 = 0.729 (P = 0.031)) and 29 flavonoids (R2 = 0.521 (P = 0.054)). Correlation coefficient (r) analysis showed that MW, log P, and MR may be positively correlated with IC50 of the fatty acids (MW, r = 0.743; log P, r = 0.642; and MR, r = 0.815) and IC50 of the flavonoids (MW, r = 0.642; log P, r = 0.495; and MR, r = 0.643). Further studies will warrant possible applications of T. molitor adults as an antioxidant.

Insect cuticle tanning (pigmentation and sclerotization) is a complex and vital process, which includes hydroxylation of initial amino acid, tyrosine, to DOPA and decarboxylation of DOPA to dopamine. In the pigmentation process, dopamine further undergoes two N-acylation reactions to yield N-acetyldopamine (NADA) and N-β-alanyldopamine (NBAD). In the former reaction, arylalkylamine N-acetyltransferase (AANAT1) converts dopamine to NADA, and in the later reaction, aspartate 1-decarboxylase (ADC) provides β-alanine, which is conjugated with dopamine catalyzed by NBAD synthase (Ebony) for production of NBAD. In this study, we performed functional genomics of TmAANAT1, TmADC and Tmebony to determine whether they are required for cuticle pigmentation in Tenebrio molitor adults. Loss of function of these genes by RNAi caused the significantly darker body color than that of control animals. Note that, although all phenotypes exhibited dark cuticle pigmentation, RNAi of either TmADC or Tmebony only altered brownish outer region of the cuticle to dark/black. In contrast, RNAi of TmAANAT1 had no effect on the brown hue of the outer cuticle layer, but less or no pigmented inner region of the cuticle became significantly darker than those of control adults. These results suggest that, like that seen in TcAANAT1- or TcADC-deficient Tribolium castaneum adults, NADA produced by a reaction by TmAANAT1 contributes the lighter inner cuticle layer(s), whereas NBAD appears to do the highly pigmented outer cuticle layer(s) of the cuticle of T. molitor adults. This work was supported by NRFs (NRF-2015R1A6A3A04060323 and NRF-2018R1A2B6005106).

IKK-γ is an essential protein to form IKK complex which regulate NF-κB. We identified TmIKK-γ (or TmKenny) gene which has 1,521 bp of nucleotides encoding 506 amino acid residues. Domain analysis of TmIKK-γ shows that there are one NF-κB essential modulator (NEMO) domain and a leucine zipper domain. Expression of TmIKK-γ gene was gradually increased from egg to 2-day-old pupal stage, dramatically decreased until 7 day-old pupal stage, and then it was gradually increased. TmIKK-γ transcripts were highly expressed in fat body and hemocytes in late instar larvae and integuments, fat body and Malpighian tubules in 5 day-old adult. TmIKK-γ was drastically induced by E. coli after 3 h challenges and by S. aureus at 3 and 12 h-post injection in hemocytes. TmIKK-γ was not induced by C. albicans although it was significantly induced by E. coli (at 3, 6 and 24 h) and S. aureus (at 9 h) in gut. In fat body, expression of TmIKK-γ was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-γ, gene specific RNAi and mortality assay was performed. larval mortality against microbial challenge was dramatically increased by TmIKK-γ RNAi. Furthermore, we investigate the tissue specific induction patterns of fourteen AMP genes in response TmIKK-γ dsRNA-treatment. In fat body, ten AMP genes out of fourteen was not significantly induced by microbial challenge in TmIKK-γ dsRNA-treated group. Based on these results, TmIKK-γ might play an important role in antimicrobial innate immune responses in Tenebrio molitor.

Tyrosinase is a key enzyme in animal melanogenic pathway that is the rate-limiting step for the production of melanin. Several synthetic and naturally occurring tyrosinase inhibitors have been studied for skin whitening, the development of natural agents is becoming more important due to the disadvantages of synthetics such as high cytotoxicity, insufficient penetration power, and low activity. The purpose of this study was to evaluate the polyphenols, antioxidant and tyrosinase inhibition activity of mealworm (Tenebrio molitor) extract and optimization of extraction condition using statically-based optimization. The results showed that extraction temperature of 92.0°C, extraction time of 36.7 min, and ethanol concentration of 79.5% provided the maximum levels of compounds of 5.69 mg GAE/g DW, tyrosinase inhibition of 78.8%, and radical scavenging activity of 100.8 μg/ml in the validation experiment.

This study was performed to evaluate the quality characteristics of functional Dasik prepared with a mixture of freeze-dried mealworm (Tenebrio molitor) powder and dried pollack powder along with assessment of the general and fatty acid compositions of mealworms. General compositions, except for moisture content of freeze-dried mealworm powder, were higher than those of raw mealworms. The ratios of saturated fatty acids and unsaturated fatty acids of freeze-dried mealworm powder and raw mealworms were 1:3.31 and 1:3.19, respectively. Amounts of oleic acid, which was the most abundant among unsaturated fatty acids, of freeze-dried mealworm powder and raw mealworms were 41.12 and 37.84%, respectively. For color values, greater content of freeze-dried mealworm powder in functional Dasik resulted in lower L and b scores, whereas a value increased. In the case of mechanical properties, greater content of freeze-dried mealworm powder resulted in significant reduction of hardness, chewiness, and gumminess. In the case of sensory evaluation, color, taste, and overall quality of DPMD50, which was prepared with a 1:1 ratio of freeze-dried mealworm powder and dried pollack, were the highest. It was concluded that DPMD50 is a nutritious combination of edible insects and fish for protein fortification for growth and the elderly.

본 연구는 고기능 미래 식량자원으로서 이용 가치가 높은 밀웜과 검은콩을 첨가한 쿠키를 제조하고, 밀웜 및 검은콩 첨가량에 따른 품질특성, 항산화 활성, 기호성을 비교해 보고자 수행되었다. 밀웜의 첨가량을 0%, 10%, 20%, 30%로 증가한 쿠키와 밀웜과 함께 검은콩 분말 10%, 20% 를 첨가한 쿠키를 제조하여, 품질특성, 항산화 활성, 관능 특성을 비교한 결과는 다음과 같다. 팽창율, 손실율은 밀웜만 첨가한 쿠키와 밀웜과 검은콩 분말을 함께 첨가한 쿠키 모두에서 밀웜 분말 첨가량이 증가함에 따라 유의적으로 증가하였다. 총 폴리페놀 함량, DPPH 라디칼 소거능, FRAP, ABTS 측정 결과 밀웜 분말과 검은콩 분말을 첨가한 쿠키 모두에서 아무것도 첨가하지 않은 쿠키보다 활성이 높게 측정되었다. 밀웜 분말의 첨가량이 증가할수록 항산화 활성은 증가하여 밀웜 분말 30% 첨가군에서 가장 높은 항산화 활성을 나타내었다. 또한 동일한 양의 밀웜에 검은콩 분말을 함께 첨가한 쿠키는 밀웜 분말만 첨가한 쿠키 보다 총페놀함량, DPPH 라디칼 소거능, ABTS 라디칼 소거능이 높게 측정되었다. 밀웜만 첨가한 쿠키의 색도는 첨가량이 증가할수록 쿠키의 밝기(L 값)와 황색도(b 값)는 유의적으로 감소하고, 적색도는(a 값) 증가 하였고, 밀웜과 검은콩을 함께 첨가한 쿠키는 적색도(a 값), 밝기 (L 값), 황색도 (b 값)가 모두 감소하는 것으로 측정 되었다. 쿠키의 조직감 측정결과 밀웜 첨가량이 증가할수록 쿠키의 강도가 유의적으로 감소하였으나(p < 0.05), 검은콩을 함께 첨가한 쿠키는 강도가 강하게 나타나 밀웜 증가에 의한 쿠키 강도의 감소를 검은콩을 첨가함으로써 개선할 수 있었다. 이상의 결과에서 밀웜 쿠키의 품질특성, 항산화 활성, 기호성을 모두 향상하기 위해서는 밀웜 분말을 10% 첨가하여 제조하는 것이 가장 우수하다는 것 을 알 수 있었다.

This study aimed to investigate quality characteristics of white bread with Tenebrio molitor L. powder (TMP). Because of the proximate composition analysis of the white bread with TMP, the content of protein and crude fat increased slightly in proportion to brown dough, and ash was highest in white breads with 7% added TMP. The bread volume decreased as TMP content increased. The control bread lightness was highest at 78.3, and was negatively correlated with the TMP content. The degree of redness increased with the TMP concentration. The total free amino acid contents were in the following order: white bread with 7% TMP > white bread with 5% TMP > white bread with 3% TMP. With respect to the texture, white bread springiness and cohesiveness with TMP decreased by storage period. The chewiness and brittleness of white breads with TMP decreased significantly with higher concentrations. With extended storage, the bread with TMP added showed lower springiness and cohesiveness; while bread with a lower TMP percentage maintained better chewiness and brittleness. The sensory taste preference was highest for white bread with 3% TMP. Residual appetite scores showed a lower taste preference as TMP content increased and was lowest for white bread with 3% TMP.