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Sustainable expansion of the insect farming industry requires valorization of the generated frass, a by-product with fertilizer and soil improver potential. Commercialization is currently restricted by Regulation (EU) No 2021/1925, considering insect frass as animal manure and imposing a pathogen-reducing heat treatment. This study investigated the microbiological safety of frass from industrial black soldier fly larvae (BSFL) and yellow mealworm (YM) production, untreated and hygienized by drying, pelleting or composting. Results showed that despite high microbial counts for both viable cells and aerobic endospores, Salmonella spp. were absent in untreated frass, Escherichia coli counts were low compared to other manure types, while the number of Enterococci was similar. The hygienisation procedures induced little change in the total number of viable cells and aerobic endospores and exerted more impact on temperature-sensitive Enterobacteriaceae compared to more tolerant Enterococcaceae. Drying, pelleting and composting were not equally suited for hygienization of frass from both insect species due to differences in initial microbial load and physical composition. Generally, it could be concluded that YM frass will meet the EU microbiological criteria more easily than BSFL frass and may even do so without hygienization. In case hygienization is necessary, reduction of the number of E. coli by the applied treatment will be the factor determining regulation compliance. Compared to Enterococcaceae, E. coli abundance in frass is lower and the species is more efficiently reduced. Oven drying is suited for both BSFL and YM frass but is expensive as it requires an external heat source. Pelleting seems applicable to YM frass, BSFL frass requires preliminary heating to reduce its moisture content. Composting of BSFL frass may produce a safe product if temperatures reached throughout the process are high enough. Composted YM frass can be considered microbiologically more unstable/unsafe than untreated YM frass, due to water addition at the process start. It may be concluded that flexibility in treatment procedures for insect frass, aimed for use as fertilizer, can be allowed. Microbiological safety is ensured after reference treatment (70 °C–60 min) but can also be achieved after milder processing or even without treatment, which would reduce costs and preserve frass quality. Graphical abstract

Trichogramma spp., among the most common parasitoids used for augmentation biological control, often are mass-reared on eggs of the Mediterranean flour moth, Ephestia kuehniella (Zeller). To evaluate removal of nutritional components from the E. kuehniella larval diet and reduce production costs, colonies were maintained using one of three diets: a standard diet consisting of eight ingredients, a reduced diet containing whole wheat flour, glycerol, and Brewer’s yeast, or a third minimal diet of only whole wheat flour. The standard diet sustained the fastest larval development, female pupae with the greatest mass, the highest level of adult emergence, and production of the most eggs per female. Eggs from moths reared as larvae on the standard or reduced diet had equivalent mass, length, and percent hatch. Females from larvae fed the minimal diet produced eggs with the least mass that were shorter and had the lowest percent hatch. Eggs from the three E. kuehniella colonies were exposed separately to Trichogramma brassicae Bezdenko females to determine their acceptance for oviposition. More of the eggs from the standard diet were parasitized by the females, eggs from the reduced and minimal diets being less acceptable. The percent emergence of the parasitoids was the same regardless of diet; however, the largest wasps emerged from the standard diet eggs and a greater proportion of them were females. Consequently, the standard E. kuehniella larval diet resulted in the highest rate of reproduction and robust eggs that produced superior T. brassicae wasps.

Laricobius nigrinus (Coleoptera: Derodontidae) Fender and Laricobius osakensis (Coleoptera: Derodontidae) Montgomery and Shiyake have been mass produced by Virginia Tech as biological control agents for the hemlock woolly adelgid (HWA), Adelges tsugae (Hemiptera: Adelgidae) Annand, for the past 15 and 9 yr, respectively. Herein, we describe modifications of our rearing procedures, trends and analyses in the overall production of these agents, and the redistribution of these agents for release to local and federal land managers. Based on these data, we have highlighted three major challenges to the rearing program: 1) high mortality during the subterranean portion of its life cycle (averaging 37% annually) reducing beetle production, 2) asynchrony in estivation emergence relative to the availability of their host HWA minimizing food availability, and 3) unintended field collections of Laricobius spp. larvae on HWA provided to lab-reared larvae complicating rearing procedures. We further highlight corresponding avenues of research aimed at addressing each of these challenges to further improve Laricobius spp. production.

Development of a successful in vitro rearing protocol has been essential for pesticide safety assessment of immature honey bee workers under laboratory conditions. In contrast, pesticide safety testing of honey bee drones is limited, in part due to the lack of successful laboratory rearing protocols for this reproductive caste. Considering that healthy drones are essential for successful mating and reproduction of the honey bee queen, a standardized in vitro rearing protocol for honey bee drones is necessary to support reproductive safety studies, as well as to gain a deeper understanding of honey bee drone development. Using the established in vitro rearing protocol for honey bee workers, we modified the days of grafting and pupal transfer, as well as the diet volume, pupation plate orientation, and absorbent tissue in the pupal wells to successfully rear honey bee drones in vitro. In vitro-reared drones were evaluated for gross wing abnormalities, body weight, testes weight, and abdominal area, and compared with age-matched drones reared in field colonies. We found that honey bee drones reared in a vertically oriented pupation plate containing WypAll.sup.® absorbent tissue in each well had a mean survival to adulthood of 74 ± 3.5% (SEM) until adulthood. In contrast, drones reared in a horizontally oriented pupation plate containing Kimwipe.sup.® absorbent tissue in each well had significantly lower survival (5.5 ± 2.3%) and demonstrated gross wing abnormalities. All in vitro-reared drones had significantly lower body weight, testes weight and abdominal area relative to colony-reared control drones. Accordingly, we successfully developed an in vitro rearing protocol for honey bee drones which has the potential to improve future reproductive safety assessment of pesticides for honey bees.

Lilioceris egena (Weise) (Coleoptera: Chrysomelidae) is an approved classical biological control agent that feeds on the aerial reproductive structures (i.e., bulbils) of the invasive air potato vine in Florida. The Florida Department of Agriculture and Consumer Services, Division of Plant Industry, has mass-produced and released over 100,000 beetles since 2021. Mass rearing can be challenging for highly specialized herbivores like L. egena. Therefore, we present a detailed protocol to mass rear L. egena under laboratory settings. This protocol ensures consistent production of approximately 675 beetles per month/colony for research and field applications while maintaining efficiency and affordability. Additionally, we determined how the number of mating pairs and the condition of air potato bulbils (whole or bisected) used in the mass rearing system influenced the number of adults produced. Results showed that 17 mating pairs per bulbil (of approximately 150 g) was an optimal beetle density to maximize beetle production and maintain a healthy colony. The condition of bulbils utilized did not influence the number of adults produced. This protocol can be adopted by integrated pest management (IPM) and master gardener programs, universities, and regulatory agencies in other southeastern states currently facing air potato infestations.

Mass Production of Beneficial Organisms: Invertebrates and Entomopathogens is an essential reference and teaching tool for researchers in developed and developing countries working to produce \"natural enemies\" in biological control and integrated pest management programs.As we become aware of the negative impact of pesticides in human health and.

Louse-borne diseases have caused millions of deaths around the world and are currently re-emerging in some countries. Implementing a simple and accessible lice-rearing system would significantly advance research on the louse–pathogen cycle and vectorial capacities. Several in vivo and in vitro lice-rearing methods have been developed. However, these approaches have drawbacks, making lice production more difficult. Here, we aimed to adapt the Orlando (Or) strain of body lice on an artificial membrane. The Hemotek system and a Petri dish system covered with a Parafilm membrane were tested on newly hatched first-stage larvae (L1). Rearing follow-up consisted of recording dead, fed and moulted specimens throughout the experiments. In addition, microscopic ultra-structures, blood meal digestion and sterility were evaluated and compared to those of larvae being reared on rabbit hosts. When using heparinised blood on a Petri dish, we were able to maintain one generation of body lice. Development into adulthood was recorded 21 days after hatching, and 52 eggs were deposited. Inspection of the blood meal revealed a colour difference among lice fed in vitro and in vivo, while microscopic investigations did not show any differences. The in vitro lice-rearing experiments were conducted in accordance with animal welfare requirements and therefore have the potential to replace animal models in various biological assays. Human lice are the only hematophagous ectoparasites specific to human hosts. They transmit epidemic typhus, trench fever and relapsing fever, diseases which have already caused millions of deaths worldwide. In order to further investigate lice vectorial capacities, laboratory-controlled live lice colonies are essential. Previously developed lice-rearing methods significantly advanced research on louse-borne diseases and louse biology. In this study, we aimed to develop a rearing technique for the Orlando (Or) strain of body lice on an artificial membrane. We tested two systems, namely the Hemotek feeding system and a Petri dish with the lice being fed through a Parafilm membrane. Lice longevity and development were drastically affected by the blood anticoagulant. Additionally, heparinised human blood on a Petri dish was the best candidate when compared to the control group (reared on a rabbit). Therefore, this strategy was applied to 500 lice. Development into adulthood was recorded after 21 days (17 days for the rabbits), and 52 eggs were deposited (240 for the rabbits). In this study, we were able to maintain one generation of body lice on an artificial membrane with comparable feeding and longevity rates to those fed on live rabbits. However, lice fecundity decreased on the artificial membrane. In vitro lice-rearing experiments will enable pathogen infection assays and pesticide bioassays to be carried out in accordance with animal welfare requirements.

The mass production of insects for food and feed is an expanding North American industry. Facilities that mass rear insects are at risk of pest infestations because the optimal environmental conditions for rearing beneficial species may also support the development of pest species. Here, we present the first recorded results detailing the interactions between dermestids and farmed crickets. Cricket farms have reported extremely low harvest yield during heavy dermestid infestations, but the exact reasons for this low yield are unknown. Many dermestid larvae are covered in dense, detachable, barbed setae called hastisetae, which are used by the larvae as an active trapping system against arthropod predators. We designed a series of experiments to test the hypotheses that a dermestid pest of cricket farms, black larder beetle (Dermestes ater DeGeer (Coleoptera: Dermestidae)), may be directly impacting Gryllodes sigillatus Walker (Orthoptera: Gryllidae) yield through the physical effects of hastisetae ingestion and/or indirectly impacting cricket yield through competition for fishmeal, a primary source of protein in conventional cricket feed. Our predictions that G. sigillatus life history and survival would be negatively affected by dermestids were largely refuted. Females fed infested diets grew less mass, but not smaller body size, compared to females fed uninfested diets. We also found that while G. sigillatus experienced delayed growth early in life after living with dermestids, they were able to tolerate living with, and consuming, dermestid larvae. We discuss how these findings have led to new hypotheses concerning how dermestid infestations drive reductions in cricket farm yield. Key words: insects as food and feed, interaction, insect rearing, competition

Over the past 2 decades, the potential of insects as food and feed has been recognized globally. Insects as feed ingredients can improve sustainability because of their lower greenhouse gas emissions and their potential to transform organic wastes into high-quality feed rich in nutrients. However, currently, the practical use of insects as food or feed is limited by the high costs of insect production. A great deal of effort is required to improve the rearing technology necessary to establish the principles of insect farming. Several insect species have become industrialized using existing methods of production. The most common industrialized insect species intended as feed and food include the yellow mealworm, Tenebrio molitor L. (Coleoptera:Tenebrionidae); the house cricket, Acheta domesticus L. (Orthoptera: Gryllidae); the black soldier fly, Hermetia illucens L. (Diptera: Stratiomyidae); and the house fly, Musca domestica L. (Diptera: Muscidae). This review focuses on describing the existing rearing methods for these 4 insect species, which may provide a basis for future research to enhance insect production capabilities.

Edible insects offer a viable alternative protein source to help meet the protein demands of a growing population. Optimizing insect mass-rearing for food and feed production depends on an understanding of insect life history. However, supporting data on growth, development, and reproduction from hatch to adulthood is often not available for many farmed insects, such as the decorated cricket (Gryllodes sigillatusWalk.). Here, we describe the life history of G. sigillatus from hatch to adulthood at 30 °C for traits relevant for mass-rearing and colony management. Female crickets first reached adulthood after 29 d and weighed 292.0 mg ± 74.09 mg, and male crickets first reached adulthood after 35 d and weighed 200.96 mg ± 34.51 mg. Crickets had 7 nymphal instars most characterizable by head width. Sex was identified by the development of ovipositors in females, and wings in both sexes. Crickets oviposited 56.74 ± 31.77 eggs every 48 h over 30 d and eggs hatched after 10.6 ± 0.5 d. This information provides the foundation to start and manage a cricket colony, to conduct research on life history and performance, and to facilitate practitioners to make informed decisions about rearing practices or identify arising issues. We highlight ways that a fundamental understanding of cricket biology can be informative for optimizing cricket growth, reducing variability in yield, and informing future precision farming practices.