We introduce a method for preserving yellow mealwom (Tenebrio molitor) larvae for an extended period and show that a high percentage of larvae can survive in good health under low-temperature storage conditions combined with specific diapause termination conditions. When storing larvae for 140 days, the storage temperature can be varied based on our goals, giving us control over yellow mealworm production to meet specific demands. To produce adult beetles, storing larvae at 15 ℃ with wheat bran and ending diapause at 30 ℃ resulted in 90% pupation rate, with 60% becoming adults in 21 days. If our aim is larvae production, storing them at 10–12 ℃ with wheat bran and ending diapause at 25–30 ℃ allows the larvae to reach a suitable weight for processing. This approach ensures long-term storage of yellow mealworm larvae and provides a practical way to control their development, allowing efficient mass production tailored to market demands.
The yellow mealworm beetle, Tenebrio molitor L. (Coleoptera: Tenebrionidae), has long been used as a key study organism in many fundamental researches, including biochemistry, physiology, and behavior. Lifespan and reproduction are two of the most important components of fitness in all insects, but it remains largely unexplored how these two traits are influenced by macronutrient intake in this beetle. In this study, we used the nutritional geometry framework to analyze the complex and interactive effects of dietary protein and carbohydrate intake on lifespan and reproductive performance in T. molitor beetle. Lifespan and the number of eggs laid throughout the lifetime were quantified from more than 2,000 individual beetles provided with one of 35 chemically defined diets representing a full combination of seven protein-to-carbohydrate ratios (P:C= 0:1, 1:5, 1:2, 1:1, 2:1, 5:1, or 1:0) and five protein plus carbohydrate concentrations (P+C=25.2, 33.6, 42, 50.4, or 58.8 %, dry mass). All measures of lifespan and egg production were expressed highly at high caloric intake, but they differed in the optimal P:C ratio where traits peaked. While lifespan was the longest at a moderately carbohydrate-biased P:C ratio of 1:1.36, the rate of egg production was maximized at a protein-biased P:C ratio of 1.75:1, suggesting a possible nutrient-mediated trade-off between lifespan and daily reproductive efforts in T. molitor beetles. Lifetime egg production was maximized at a P:C ratio of 1.31:1, which was still protein-biased but lower than that maximized egg production rate. Reproductive lifespan was the longest at a P:C ratio of 1:1.06. When given a food choice, T. molitor beetles preferred a P:C ratio of 1:1, which is closest to the ratio that enables T. molitor beetles to stay reproductively active as long as possible.
The yellow mealworm, Tenebrio molitor L. (Coleoptera: Tenebrionidae), is an important industrial insect commercially produced around the world as food and feed. Temperature and nutrition are the two most influential environmental factors determining the rearing conditions in insects, but little is known about how these two factors interact to affect the performance of T. molitor larvae. In this study, we investigated the combined effects of temperature and dietary protein:carbohydrate (P:C) ratio on key performance traits in T. moltior larvae. Throughout their larval stage, the insects were reared on one of 36 treatment combinations of six temperatures (19, 22, 25, 28, 31, 34 °C) and six protein:carbohydrate ratios (P:C = 1:5, 1:2, 1:1, 2:1, 5:1, 1:0) and their survivorship, development, growth rate, and pupal mass were monitored. Survivorship was high at low temperatures (< 25°C) and high P:C ratios (>1:1), but decreased with increasing temperature and decreasing P:C ratio. Increase in rearing temperature accelerated larval development but resulted in a reduced pupal mass. Thermal optimum for pupal mass (19.3°C) was thus lower than that for development time (28.1°C). The growth rate was maximized at 27.9°C and P:C 1.65:1 and decreased as both the temperature and the P:C ratio deviated from their optimum. All four key performance traits (survivorship, development time, pupal mass, growth rate) were optimized at temperatures between 25.7 and 27.4°C and P:C ratios between 1.17:1 and 2.94:1. Our data provide insights into how the production and nutritional value of T. molitor larvae can be improved through adjusting their rearing conditions.
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).