Covert RNA viruses in medflies differ in their mode of transmission and tissue tropism.

Numerous studies have demonstrated the presence of covert viral infections in insects. These infections can be transmitted in insect populations via two main routes: vertical from parents to offspring, or horizontal between nonrelated individuals. Thirteen covert RNA viruses have been described in the Mediterranean fruit fly (medfly). Some of these viruses are established in different laboratory-reared and wild medfly populations, although variations in the viral repertoire and viral levels have been observed at different time points. To better understand these viral dynamics, we characterized the prevalence and levels of covert RNA viruses in two medfly strains, assessed the route of transmission of these viruses, and explored their distribution in medfly adult tissues. Altogether, our results indicated that the different RNA viruses found in medflies vary in their preferred route of transmission. Two iflaviruses and a narnavirus are predominantly transmitted through vertical transmission via the female, while a nodavirus and a nora virus exhibited a preference for horizontal transmission. Overall, our results give valuable insights into the viral tropism and transmission of RNA viruses in the medfly, contributing to the understanding of viral dynamics in insect populations. IMPORTANCE: The presence of RNA viruses in insects has been extensively covered. However, the study of host-virus interaction has focused on viruses that cause detrimental effects to the host. In this manuscript, we uncovered which tissues are infected with covert RNA viruses in the agricultural pest Ceratitis capitata, and which is the preferred transmission route of these viruses. Our results showed that vertical and horizontal transmission can occur simultaneously, although each virus is transmitted more efficiently following one of these routes. Additionally, our results indicated an association between the tropism of the RNA virus and the preferred route of transmission. Overall, these results set the basis for understanding how viruses are established and maintained in medfly populations.

Beauveria australis finds a new host in French dung beetles introduced to Australia.

Exotic dung beetles have been introduced to Australia for over 50 years to mitigate issues caused by dung produced by livestock. This study aims at identifying fungi affecting a beetle colony and investigating their source. Fungal hyphae emerging from the cuticle of dead beetles were cultured and a multigene phylogeny showed that Beauveria bassiana and B. australis both infected these introduced beetles, likely acquired through local soil or dung. This is the first record of B. australis infecting family Scarabaeidae. This study highlights the importance of fungal outbreak management in insect rearing before release, and challenges associated with new pathogens.

Egg development and viability in three species of Cyclocephala (Coleoptera: Scarabaeidae: Dynastinae).

Different species of Cyclocephala scarab beetles (Scarabaeidae, Dynastinae) perform key functional roles in both natural and agricultural systems, such as the cycling of organic matter and pollination, while also being known as destructive pests both as immatures and adults. Therefore, the identification of biological parameters is crucial for defining strategies for their conservation and efficient pest management. In a forest fragment within the Brazilian Atlantic Forest biodiversity hotspot, we field-captured adult individuals of Cyclocephala cearae, C. celata, and C. paraguayensis then reared and bred them under controlled temperature and humidity conditions. On a daily basis, we individually weighted eggs of all three species, from oviposition until hatching, and monitored egg development parameters (i.e., incubation duration, viability, and egg weight increase). Our findings provide novel empirical evidence showing (i) a positive correlation between egg weight and incubation duration, (ii) idiosyncratic characteristics on egg development, and (iii) a negative (involuntary) effect of manipulation on egg development and viability. Thus, the successful breeding and rearing of Cyclocephala spp. is correlated with egg integrity and the targeted species. Our analyses present a quantitative understanding of the egg phase and can assist in refining strategies for ovicidal activity and pest management of Cyclocephala spp. in agriculture systems. Moreover, they can provide a basis for new studies related to captivity breeding, pollinator management, and developmental biology for biodiversity conservation.

Quality control applications for recovering an inbred colony of Bagrada hilaris (Hemiptera: Pentatomidae).

The invasive stink bug, Bagrada hilaris (Burmeister), recently became established in the southwestern United States and has become a major pest of broccoli and other cole crops. Due to concerns about its possible establishment in Florida, a colony of this pest was maintained in quarantine to conduct research on its environmental requirements. The colony was reared reliably with approximately 300 adults per generation but began to decline in generation 16. Due to unknown causes, only about 73 females were recovered to mate and oviposit during the final 46 days. However, a corresponding decrease in the number of mated pairs did not reduce the yield of eggs, nymphs, and adults per day, but the females were maintained for fewer than the normal 160 days per generation. Therefore, quality control procedures were implemented to increase the number of days the colony produced adults in subsequent generations. The goal of producing approximately 400 adults per generation was accomplished during 104, 160, and 156 days, respectively, in generations 17, 18, and 19. The purpose of this research was to develop quality control procedures for rearing B. hilaris, use the procedures to restore a colony in quarantine, and describe how quality control can be used to maintain small colonies of insects. Implementing quality control procedures when a colony is established can help to prevent its decline.

Harnessing thermal plasticity to enhance the performance of mass-reared insects: opportunities and challenges.

Insects are mass-reared for release for biocontrol including the sterile insect technique. Insects are usually reared at temperatures that maximize the number of animals produced, are chilled for handling and transport, and released into the field, where temperatures may be considerably different to those experienced previously. Insect thermal biology is phenotypically plastic (i.e. flexible), which means that there may exist opportunities to increase the performance of these programmes by modifying the temperature regimes during rearing, handling, and release. Here we synthesize the literature on thermal plasticity in relation to the opportunities to reduce temperature-related damage and increase the performance of released insects. We summarize how and why temperature affects insect biology, and the types of plasticity shown by insects. We specifically identify aspects of the production chain that might lead to mismatches between the thermal acclimation of the insect and the temperatures it is exposed to, and identify ways to harness physiological plasticity to reduce that potential mismatch. We address some of the practical (especially engineering) challenges to implementing some of the best-supported thermal regimes to maximize performance (e.g. fluctuating thermal regimes), and acknowledge that a focus only on thermal performance may lead to unwanted trade-offs with other traits that contribute to the success of the programme. Together, it appears that thermal physiological plasticity is well-enough understood to allow its implementation in release programmes.

Gross Morphology of Diseased Tissues in Nezara viridula (Hemiptera: Pentatomidae) and Molecular Characterization of an Associated Microsporidian.

Nezara viridula (L.) (Hemiptera: Pentatomidae), commonly known in the U.S. as the southern green stink bug (SGSB), is a cosmopolitan, highly polyphagous feeder that causes severe damage to a wide range of agronomically important crops such as fruit, vegetable, grain, tobacco, and cotton, throughout much of the United States, and is a global pest of considerable ecological, agricultural, and economical interest. During dissection of female Nz. viridula, conspicuous black and brown spots or lesions were observed on various internal organs. To determine the cause of these spots or lesions, tissues of fat body, spermatheca, ovaries, and ovulated eggs were collected from healthy and infected individuals. The gross morphology of the spots was characterized, and the microorganisms associated with the infection were identified by amplicon sequencing of the V4 region of the small subunit rRNA gene. The presence of a microsporidian pathogen Nosema maddoxi, Becnel, Solter, Hajek, Huang, Sanscrainte, & Estep (Microsporidia: Nosematidae) which has been observed on other species of stink bug, was evidenced for the first time. The characterization of the gross morphology of this associated microsporidian may enable more rapid determination of microsporidia infection in stink bug colonies and field populations.

Effects of flaxseed cake fortification on bread shelf life, and its possible use as feed for Tenebrio molitor larvae in a circular economy: preliminary results.

BACKGROUND: Bread represents a significant share of food waste worldwide. The extension of the bread shelf life together with innovative systems of food waste treatment might decrease waste biomass decay, the need for transportation, and the need for storage. In recent years, insects have been selected as a valuable tool for food waste treatment owing to their capability to transform low-value food waste into biomass with high nutritional value. Bakery wastes can be used profitably for this purpose. This work had two objectives: (i) to measure the impact of flaxseed cake fortification on bread shelf life depending on the leavening agent (baker's yeast vs sourdough); (ii) to evaluate the possible reuse of the stale bread fortified with flaxseed cake for Tenebrio molitor rearing. RESULTS: Our results showed that fortification seemed to slow the hardening rate of bread, particularly if baker's yeast was used. The time necessary for mold to appear in sourdough bread doubled (from 2 to 4 days). The addition of flaxseed cake to the recipe determined an increase of its scrap consumption by T. molitor larvae. We also observed a significant increase in the body mass of the T. molitor larvae fed with bread obtained with the brewer's yeast with respect to larvae fed with the sourdough. CONCLUSION: Taken together, these preliminary data can indicate that sourdough bread fortified with 5% of flaxseed cake can represent a promising tool to reduce food waste and to recycle bread scraps by a novel zero-waste approach. © 2021 Society of Chemical Industry.

Historic Assessment and Analysis of the Mass Production of Laricobius spp. (Coleoptera: Derodontidae), Biological Control Agents for the Hemlock Woolly Adelgid, at Virginia Tech.

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.

Almond by-product composition impacts the rearing of black soldier fly larvae and quality of the spent substrate as a soil amendment.

BACKGROUND: Insect biomass is a sustainable alternative to traditional animal feeds, particularly when insects are produced on low-value high-volume agricultural by-products. Seven samples of almond by-product (hulls and shells) were obtained from processors in California and investigated for larvae production. Experiments were completed with and without larvae and spent substrate samples were assessed for their potential as soil amendments based on standard compost quality indicators. RESULTS: On average, specific larvae growth and average larval harvest weight were 158% and 109% higher, respectively, when larvae were reared on Monterey and pollinator hulls compared to nonpareil hulls and mixed shells. Larvae methionine and cystine contents were highest when larvae were reared on Monterey hulls and mixed shells, respectively. Available phytonutrients in spent substrate were affected by feedstock sample and larvae rearing. Spent nonpareil substrate without larvae had the highest NH4 -N levels and spent pollinator substrate incubated without larvae had the highest PO4 -P levels. Spent mixed shell substrate had the lowest availability of phytonutrients. CONCLUSION: The findings demonstrate that by-product composition has a significant impact on larvae growth and the properties of the spent substrate, and that spent substrate from larvae rearing requires further stabilization before application as a soil amendment. © 2020 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Olive fruit fly rearing procedures affect the vertical transmission of the bacterial symbiont Candidatus Erwinia dacicola.

BACKGROUND: The symbiosis between the olive fruit fly, Bactrocera oleae, and Candidatus Erwinia dacicola has been demonstrated as essential for the fly's larval development and adult physiology. The mass rearing of the olive fruit fly has been hindered by several issues, including problems which could be related to the lack of the symbiont, presumably due to preservatives and antibiotics currently used during rearing under laboratory conditions. To better understand the mechanisms underlying symbiont removal or loss during the rearing of lab colonies of the olive fruit fly, we performed experiments that focused on bacterial transfer from wild female flies to their eggs. In this research, eggs laid by wild females were treated with propionic acid solution, which is often used as an antifungal agent, a mixture of sodium hypochlorite and Triton X, or water (as a control). The presence of the bacterial symbiont on eggs was evaluated by real-time PCR and scanning electron microscopy. RESULTS: DGGE analysis showed a clear band with the same migration behavior present in all DGGE profiles but with a decreasing intensity. Molecular analyses performed by real-time PCR showed a significant reduction in Ca. E. dacicola abundance in eggs treated with propionic acid solution or a mixture of sodium hypochlorite and Triton X compared to those treated with water. In addition, the removal of bacteria from the surfaces of treated eggs was highlighted by scanning electron microscopy. CONCLUSIONS: The results clearly indicate how the first phases of the colony-establishment process are important in maintaining the symbiont load in laboratory populations and suggest that the use of products with antimicrobial activity should be avoided. The results also suggest that alternative rearing procedures for the olive fruit fly should be investigated.

Improvements in the Rearing of the Tachinid Parasitoid Exorista larvarum (Diptera: Tachinidae): Influence of Adult Food on Female Longevity and Reproduction Capacity.

Exorista larvarum (L.), a polyphagous gregarious larval parasitoid of lepidopterans, can be mass produced both in vivo, using the greater wax moth Galleria mellonella (L.) (Lepidoptera: Pyralidae) as a factitious host, and in vitro, on artificial media composed of crude components. The present study was focused on another aspect of E. larvarum rearing, namely the influence of adult food on parasitoid performance. The standard food, consisting of lump sucrose and cotton balls soaked in a honey and water solution (1), was compared with other foods or food combinations, namely lump sucrose alone (2), honey and water solution (3), sucrose and water solution either alone (4) or combined with bee-collected pollen (5), and, finally, pollen alone (6). All foods were provided together with distilled water supplied in drinking troughs. Based on the parameters considered (i.e., female longevity, number of eggs laid on host larvae, puparia obtained from eggs, and adults emerged from puparia), pollen alone was deemed to be the most suitable food for adult females of E. larvarum. In particular, the pollen showed a longevity-promoting effect, increasing the number of eggs laid on host larvae throughout the female lifespan. The use of this adult food may also result in a higher flexibility of the management of E. larvarum colonies because it can be replaced weekly, as no desiccation or mold infections were ever found to occur.

Body Mass Increase Induced by Eight Years of Artificial Selection in the Yellow Mealworm (Coleoptera: Tenebrionidae) and Life History Trade-offs.

Efforts to improve rearing conditions of Tenebrio molitor L. (Coleoptera: Tenebrionidae) for insect biomass production included selecting for larger size pupae. The effects of an 8-yr continuous selection of T. molitor pupae for larger size were studied. Data consisting of daily counts and weights of pupae were analyzed using regression to determine the effects of selection over time. A preliminary evaluation of food conversion, growth, fecundity, and larval survival was done to compare ancestral versus selected strains. A significant positive correlation was identified between pupal size and time indicating a significant increase in pupal size over time in the selected T. molitor strain. A preliminary comparison of ancestral and selected strains showed significantly larger pupal size, growth rate, fecundity, and efficiency of conversion of ingested food in the selected strain. However, the selected strain also showed significantly lower larval survival than the ancestral strain. The low larval survival impacted the overall productivity of the selected strain resulting in no significant differences in biomass production when compared with the ancestral strain. The potential of using selection to improve biomass productivity in T. molitor is discussed.

Microbial Dynamics during Industrial Rearing, Processing, and Storage of Tropical House Crickets (Gryllodes sigillatus) for Human Consumption.

In this study, the microbiota during industrial rearing, processing, and storage of the edible tropical house cricket, Gryllodessigillatus, was investigated. To this end, we analyzed samples from the cricket feed, obtained before feeding as well as from the cages, and from the crickets during rearing, after harvest, and after processing into frozen, oven-dried, and smoked and oven-dried (smoked/dried) end products. Although the feed contained lower microbial numbers than the crickets, both were dominated by the same species-level operational taxonomic units, as determined by Illumina MiSeq sequencing. They corresponded, among others, to members of Porphyromonadaceae, Fusobacterium, Parabacteroides, and Erwinia The harvested crickets contained high microbial numbers, but none of the investigated food pathogens Salmonella spp., Listeria monocytogenes, Bacillus cereus, or coagulase-positive staphylococci. However, some possible mycotoxin-producing fungi were isolated from the crickets. A postharvest heat treatment, shortly boiling the crickets, reduced microbial numbers, but an endospore load of 2.4 log CFU/g remained. After processing, an increase in microbial counts was observed for the dried and smoked/dried crickets. Additionally, in the smoked/dried crickets, a high abundance of a Bacillus sp. was observed. Considering the possible occurrence of food-pathogenic species from this genus, it is advised to apply a heat treatment which is sufficient to eliminate spores. Nevertheless, the microbial numbers remained constant over a 6-month storage period, whether frozen (frozen end product) or at ambient temperature (oven-dried and smoked/dried end products).IMPORTANCE The need for sustainable protein sources has led to the emergence of a new food sector, producing and processing edible insects into foods. However, insight into the microbial quality of this new food and into the microbial dynamics during rearing, processing, and storage of edible insects is still limited. Samples monitored for their microbiota were obtained in this study from an industrial rearing and processing cycle. The results lead first to the identification of process steps which are critical for microbial food safety. Second, they can be used in the construction of a Hazard Analysis and Critical Control Points (HACCP) plan and of a Novel Food dossier, which is required in Europe for edible insects. Finally, they confirm the shelf-life period which was determined by the rearer.

Microbial dynamics during production of lesser mealworms (Alphitobius diaperinus) for human consumption at industrial scale.

In this study, the microbial dynamics during an industrial production cyle of lesser mealworms (Alphitobius diaperinus), sold for human consumption, were characterised. The microbial numbers as well as the microbial diversity were generally higher for the substrate, existing of remaining feed, faeces and exuviae, than for the larvae. Most of the species-level operational taxonomic units, identified using Illumina MiSeq sequencing, that were present in the feed were also detected in the larvae and vice versa. However, bacterial diversity decreased in the larvae during rearing. These results suggested that the feed is an important determinant of the insect bacterial community, but that some bacterial species show a competitive advantage inside the insect gut and become dominant. A blanching treatment of the larvae after harvest reduced most microbial counts, but the number of aerobic endospores remained at 4.0 log cfu/g. Whereas food pathogens Salmonella spp., Listeria monocytogenes, Bacillus cereus or coagulase-positive staphylococci were not detected in our study, fungal isolates corresponding to the genera Aspergillus and Fusarium were recovered. Therefore, it cannot be excluded that mycotoxins were present. The results of this study contribute to a better understanding of the microbial dynamics and food safety aspects during the production of edible insects.

Rearing Larvae of the Avian Nest Parasite, Philornis downsi (Diptera: Muscidae), on Chicken Blood-Based Diets.

Captive rearing of insect pests is necessary to understand their biology and to develop control methods. The avian nest fly, Philornis downsi Dodge and Aitken, is a blood-sucking parasite during its larval stage and a serious threat to endemic birds in the Galapagos Islands where it is considered invasive. In order to procure large numbers of flies for biological studies, rearing media and diets were trialed for rearing the larval stage of P. downsi under controlled conditions in the absence of its avian host. P. downsi eggs were obtained from field-caught female flies, and once eggs hatched they were reared on chicken blood for the first 3 d. Following this, three diets were tested on second- and third-instar larvae: 1) chicken blood only; 2) chicken blood, hydrolyzed protein and dried milk powder; and 3) chicken blood, hydrolyzed protein and brewer's yeast. Out of 385 P. downsi larvae tested, we were able to rear 50 larvae to the adult stage. The highest level of mortality was found in the first-instar larvae. Survivorship of second- and third-instar larvae was similar irrespective of diet and diet did not significantly influence larval or pupal development times; though larvae fed the diet with brewer's yeast developed marginally faster. Pupal weights were similar to those of larvae that had developed on bird hosts in the field. To our knowledge, this is the first effective protocol for rearing a hematophagous parasitic avian fly from egg to adult in the absence of a living host.

Optimisation of a cheap and residential small-scale production of edible crickets with local by-products as an alternative protein-rich human food source in Ratanakiri Province, Cambodia.

BACKGROUND: The health status of the indigenous people of Ratanakiri Province, Cambodia, is significantly lower compared with that of the rest of the nation. The domestication and mass production of insects may represent a sustainable, cost effective and high quality alternative source of protein to traditional livestock. This study aimed to optimise a cheap and residential cricket breeding system based on unused wild resources. The development of crickets, Teleogryllus testaceus (Walker), under seven diets composed of taro aerial parts, young cassava leaves, young cashew leaves and brown rice flour (with or without banana slices), versus a traditionally used broiler feed diet was studied. RESULTS: Cricket mortality was low in all diets, except the two cashew-based diets. Total biomass was significantly higher under the broiler feed, in addition to the two diets containing a combination of cassava leaf powder and brown rice. Yet, crickets fed with the taro diet had the highest percentage of protein. Concerning the breeding system cost, units using cassava leaves were the cheapest. CONCLUSION: Diets based on cassava leaves seem to be the most promising. Nevertheless, to produce crickets with a high body mass and a high protein level, a new experiments must be devised in which cassava leaf maturity will be adapted to fit with the cricket growth stage. Moreover, to reduce the cost of the breeding units, handmade local products should be used instead of purchased components.

Effects of New Dietary Ingredients Used in Artificial Diet for Screwworm Larvae (Diptera: Calliphoridae).

Spray-dried whole bovine blood, dry poultry egg, and a dry milk substitute are the constituents of the artificial diet currently used for mass rearing screwworm larvae, Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae). Due to high cost and uncertainty of the commercial supply of spray-dried blood, research was conducted to identify alternative, locally available, inexpensive, dietary ingredients which could reduce cost of rearing and eliminate concerns of short supply. Experimental diets were prepared without blood component and with various ratios of bovine blood or blood cell product and defatted soy flour. Results indicate that spray-dried bovine blood can be replaced by a readily available and less expensive blood cell product. When the quantity of whole dried blood or blood cell component was reduced or removed completely from the diet, the larvae did not feed adequately, resulting in high mortality. Those larvae that survived produced pupae that were of unacceptable quality. When the milk product was replaced by soy flour, pupae were slightly smaller than those reared using the current diet; however, replacement of egg product with soy flour produced even smaller pupae. Longevity of adult flies that emerged from these small pupae was short and the females deposited few eggs. These results indicate that soy flour cannot replace the blood component from the diet, but can replace the milk product successfully. It is likely that some factor or a combination of factors in the blood act as feeding stimulants, without which larvae are unable to feed normally, resulting in high larval mortality.

First report of Oryctes rhinoceros nudivirus (Coleoptera: Scarabaeidae) causing severe disease in Allomyrina dichotoma in Korea.

Oryctes rhinoceros nudivirus (OrNV) has been known to cause severe disease in coconut palm rhinoceros beetle, Oryctes rhinoceros, in Southeastern Asia and is used as a biological control to reduce the pest population. Here, we report for the first time that the OrNV may have landed on Korea and may be the major pathogen for diseased larvae of Korean horn beetle, Allomyrina dichotoma. After peroral inoculation, over 60% of infected larvae perished in 6 wk. This viral disease spreads very fast in several locations throughout Korea. This threat not only makes economic loss of local farms rearing A. dichotoma larvae but also may disturb the ecosystem by transmitting to wild A. dichotoma.

Postegression Feeding Enhances Growth, Survival, and Nutrient Acquisition in the Endoparasitoid Toxoneuron nigriceps (Hymenoptera: Braconidae).

Toxoneuron nigriceps Viereck (Hymenoptera: Braconidae), a koinobiont endoparasitoid of the tobacco budworm, Heliothis virescens F. (Lepidoptera: Noctuidae), derives nutrition from the host hemolymph during the internal portion of its larval development but feeds destructively on host tissues externally after egression. To investigate the importance of this tissue-feeding phase, and to evaluate the behaviors associated with postegression feeding, T. nigriceps larvae were subjected to one of four treatments: 1) allowed to carry out normal tissue feeding, 2) deprived of tissue feeding, 3) presented with tissues scraped away from the host remains, and 4) fed tissues scraped from an unparasitized H. virescens larva. Additionally, total carbohydrates, lipids, and proteins were quantified from pre and posttissue feeding T. nigriceps larvae to examine the effect of postegression feeding on parasitoid nutritional physiology. Parasitoids that received no tissues after egression, or that received tissue from an unparasitized H. virescens larva, had significantly smaller body masses at all stages than those allowed to feed naturally or fed tissues scraped from a parasitized host. Parasitoids that underwent normal host feeding after egression also reached larger masses then those fed scraped host tissue. Parasitoids that received no tissue after egression survived to adulthood significantly less often than those that were presented with any H. virescens tissue. This suggests that postegression tissue feeding is a vital developmental step for T. nigriceps, and that T. nigriceps will not only feed when normal postegression behavior is disrupted, but will also feed on unparasitized tissue. The quantification of macronutrients in the tissues of pre and posttissue feeding T. nigriceps larvae showed significantly elevated proportions of proteins, lipids, and carbohydrates in the tissues of larvae that had completed feeding, with the greatest difference being in total lipids.

Nesting cavity diameter has implications for management of the alfalfa leafcutting bee (Hymenoptera: Megachilidae).

Body size influences performance in many bee species and may be influenced by nesting cavity diameter in cavity-nesting bees. Megachile rotundata (Fabricius) (Hymenoptera: Megachilidae) is a commercially-managed, solitary cavity-nesting bee. In M. rotundata body size has low heritability and is strongly influenced by the size of the larval provision and the diameter of the nesting cavity. Commercial nesting boxes have cavities that are 7 mm in diameter. Our goal was to examine the effects that nesting cavity diameter has on M. rotundata body size and performance by manipulating the size of cavities that are available for nesting. We provided bees with nesting cavities that ranged in size from 4 to 9 millimeters in 1 mm increments. To assess body size we measured mass and intertegular span. To assess performance we measured wing area, wing loading, sex, overwintering survival, pollen ball occurrence, and diapause status in the offspring. We also examined the reproductive output from the different nest cavity diameters. We found that the 8 mm cavities reared bees with the largest mass, and 4 mm cavities reared bees with the smallest mass. We determined that the 7 mm nesting cavity is optimal for offspring yield, the 8 mm nesting cavity is optimal for performance, and the 5 mm nesting cavity may be optimal for conservation efforts of other cavity-nesting bees. Based on the desired outcome of the bee managers, nest sizes differing from the standard may provide an advantage.