Larval Frass Research Articles

Bioconversion of chicken meat and bone meal by black soldier fly larvae: Effects of straw addition on the quality and microbial profile of larval frass

Meat and bone meal (MBM) is a kind of animal waste with high nutritive values. Bioconversion of MBM by black soldier fly larvae (BSFL) has great potential to obtain high-quality organic fertilizers. However, limited information is available on MBM waste manipulation to enhance BSFL frass quality. In the present study, BSFL were fed with chicken MBM containing increasing levels of rice straw (CK (0%), T1 (1%), T2 (1%), and T3 (3%)). The effects of straw addition into MBM on the quality and microbial profile of BSFL frass were evaluated. Results showed that MBM amended with straw did not significantly affect the body weight of BSFL and most of the nutrients (e.g. pH, EC, TN, TP and Na) in larval frass. Compared to other treatments, T1 sample had the highest organic matter (OM) value, implying proper straw addition could increase OM contents in frass. Fourier transform infrared (FTIR) analysis showed that straw addition might enhance the decomposition of aliphatic carbons and polysaccharides during MBM digestion process. Moreover, T1 sample had the highest microbial richness and Shannon diversity indices. It was supposed that proper straw addition in MBM helped build a more balanced diet and contributed to the BSFL gut health, consequently stimulating the gut microbe-mediated substances transformation or decomposition and promoting the microbial diversity in frass. Compared to CK, straw addition had significant influence on the abundances of Firmicutes, Bacteroidetes and Fusobacteria in frass. Elements including OM, TK and Na played important roles in shaping the microbial profile of BSFL frass.

Agrochemical contaminants in six species of edible insects from Uganda and Kenya.

Edible insects are currently promoted worldwide as an alternative animal protein source, but they are mostly still harvested from the wild where they are predisposed to contamination with agrochemicals. This study analysed six species of edible insects (Ruspolia differens, Rhynchophorus phoenicis, Schistocerca gregaria, Oryctes sp, Pachnoda ephippiata and Acanthoplus sp) collected from different habitats and/or reared in the laboratory in Kenya and Uganda for safety from agrochemical contaminants using liquid chromatography tandem mass spectrometry. The residue levels were statistically compared with the Codex Alimentarius Commission maximum residue limits (MRLs). Residues of only nine agrochemicals were detected in the insects out of 374 chemicals which were screened. The detected agrochemicals include two insecticides (aminocarb and pymetrozine), three herbicides (atraton, methabenzthiazuron and metazachlor) and four fungicides (carboxin, fenpropimorph, fludioxonil and metalaxyl). Ruspolia differens and adult Oryctes sp were free from detectable levels of any agrochemical. Whereas the pesticides residue levels in most insect samples were within maximum residue limits, some of them notably P. ephippiata from black soldier fly larval frass, R. phoenicis from oil palm and P. ephippiata from plant compost contained 2-, 8- and 49-fold higher levels of atraton, methabenzthiazuron and metazachlor, respectively, than MRLs. These findings demonstrate that edible insects may accumulate harmful residues of agrochemicals from the environment where they breed or forage, rendering them unsafe for human consumption or feeding animals. The mechanisms for possible bioaccumulation of these agrochemicals in the insects remains to be investigated. Development of methods for farming edible insects under regulated indoor conditions to ensure their safety as sources of food or feed is recommended.

Plastic biodegradation: Do Galleria mellonella Larvae Bioassimilate Polyethylene? A Spectral Histology Approach Using Isotopic Labeling and Infrared Microspectroscopy.

Environmental pollution by the nearly nonbiodegradable polyethylene (PE) plastics is of major concern; thus, organisms capable of biodegrading PE are required. The larvae of the Greater Wax Moth, Galleria mellonella (Gm), were identified as a potential candidate to digest PE. In this study, we tested whether PE was metabolized by Gm larvae and could be found in their tissues. We examined the implication of the larval gut microbiota by using conventional and axenic reared insects. First, our study showed that neither beeswax nor LDPE alone favor the growth of young larvae. We then used Fourier transform infrared microspectroscopy (μFTIR) to detect deuterium in larvae fed with isotopically labeled food. Deuterated molecules were found in tissues of larvae fed with deuterium labeled oil for 24 and 72 h, proving that μFTIR can detect metabolization of 1 to 2 mg of deuterated food. Then, Gm larvae were fed with deuterated PE (821 kDa). No bioassimilation was detected in the tissues of larvae that had ingested 1 to 5 mg of deuterated PE in 72 h or in 19 days, but micrometer sized PE particles were found in the larval digestive tract cavities. We evidenced weak biodegradation of 641 kDa PE films in contact for 24 h with the dissected gut of conventional larvae and in the PED4 particles from excreted larval frass. Our study confirms that Gm larvae can biodegrade HDPE but cannot necessarily metabolize it.

Feeding preference of insect larvae to waste electrical and electronic equipment plastics

Waste electrical and electronic equipment (WEEE) plastics not only pollute the environment, but are challenging to treat in an environmentally friendly manner. Biodegradation by insect larvae is potentially an eco-friendly method to treat WEEE plastics, but information about the feeding preference of insect larvae to WEEE plastics is lacking. In this study, a total of nine WEEE and pristine plastics were chosen to feed larvae of the following two insect species, i.e. Galleria mellonella and Tenebrio molitor. G. mellonella larvae significantly favor corresponding pristine plastics compared to two types of WEEE plastics, waste rigid polyurethane (RPU) and waste polystyrene (PS). One possible explanation is the increased chlorine or metals in the WEEE plastics measured using X-ray fluorescence spectrometer analysis. Scanning electron microscopy and Fourier transform infrared spectroscopy show that the destruction of physical structures and changes in surface functional groups were found in the two types of WEEE plastics in the larval frass, implying that the larvae partly biodegraded the plastics. Meanwhile, the powdered waste high impact polystyrene plastics (WHIPS) were ingested, but not the lumpy ones, indicating that the consumption by G. mellonella larvae is improved by the WHIPS physical modification. In addition, G. mellonella larvae presented the following decreasing preference for pristine plastics under individual-plastic-fed mode: RPU > phenol–formaldehyde resin > polyethylene (PE) > polypropylene > PS ≈ polyvinyl chloride; this is possibly due to differences in physical properties and chemical structures of the plastics; feeding preference of the larvae under multiple-plastics-fed mode is relatively consistent to that under individual-plastic-fed mode. Interestingly, the consumption by G. mellonella larvae of PE is higher than that of PS, while T. molitor larvae showed the opposite trend, implying that insect larvae have different plastics preference. The findings provide insights into biodegradation of WEEE plastics by insect larvae.

Attenuation of antibiotic resistance genes in livestock manure through vermicomposting via Protaetia brevitarsis and its fate in a soil-vegetable system

Scarab larvae (Protaetia brevitarsis) could transform large quantities of agricultural waste into compost, providing a promising bio-fertilizer for soil management. There is an urgent need to assess the risk of antibiotic resistance genes (ARGs) in soil-vegetable system with application of compost derived from P. brevitarsis larvae. We conducted a pot experiment to compare the changes of ARGs in the soil and lettuce by adding four types of manure, livestock manure (chicken and swine manure) and the corresponding larval frass. Significantly low numbers of ARGs and mobile genetic elements (MGEs) were detected in both larval frass compared with the corresponding livestock manure. Pot experiment showed that the detected numbers of ARGs and MGEs in bulk soil, rhizosphere soil, and root endophytes were significantly lower in the frass-amended treatments than the raw manure-amended treatments. Furthermore, the relative abundance of ARGs and MGEs with application of chicken-frass was significant lower in rhizosphere soil and leaf endophyte. Using non-metric multidimensional scaling analysis, the patterns of soil ARGs and MGEs with chicken-frass application were more close to those from the bulk soil in the control. Structural equation models indicated that livestock manure addition was the main driver shaping soil ARGs with raw manure application, while MGEs were the key drivers in frass-amended treatments. These findings demonstrated that application of livestock manure vermicomposting via scarab larvae (P. brevitarsis) may be at low risk in spreading manure-borne ARGs through soil-plant system, providing an alternative technique for reducing ARGs in organic waste.

Terpenes from herbivore‐induced tomato plant volatiles attract Nesidiocoris tenuis (Hemiptera: Miridae), a predator of major tomato pests

Biological control plays a key role in reducing crop damage by Tuta absoluta (Meyrick) and Trialeurodes vaporariorum (Westwood), which cause huge yield losses in tomato (Solanum lycopersicum L.). The mirid predator Nesidiocoris tenuis (Reuter) preys heavily on these pests, with satisfying control levels in tomato greenhouses. Although N. tenuis is known to be attracted to volatiles of tomato plants infested by T. absoluta and whitefly, little is known about the specific attractive compounds and the effect of prey density on the predator response. Y-tube olfactometer bioassays revealed that the attraction of N. tenuis to tomato volatiles was positively correlated with the density of T. absoluta infestation, unlike T. vaporariorum infestation. The predator was also attracted to volatiles of T. absoluta larval frass, but not to T. vaporariorum honeydew or T. absoluta sex pheromone. Among the herbivore-induced plant volatiles (HIPVs) that characterised the attractive plants infested with 20 T. absoluta larvae, olfactometer bioassays revealed that N. tenuis is attracted to the monoterpenes α-pinene, α-phellandrene, 3-carene, β-phellandrene and β-ocimene, whereas (E)-β-caryophyllene was found to repel the predator. In dose-response bioassays, the five-component blend of the attractants elicited a relatively low attraction in the predator, and removal of β-phellandrene from the blend enhanced the attraction of the predator to the resulting four-component blend, suggesting synergism among four monoterpenes. These findings suggest that a four-component blend of α-pinene, α-phellandrene, 3-carene and β-ocimene could be used as a kairomone-based lure to recruit the predator for the biological control of T. absoluta and T. vaporariorum.

Adult House Fly (Diptera: Muscidae) Response to Black Soldier Fly (Diptera: Stratiomyidae) Associated Substrates and Potential Volatile Organic Compounds Identification

Black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae), larvae are suspected to inhibit house flies, Musca domestica (L.) (Diptera: Muscidae), seeking oviposition sites. However, the mechanisms regulating these interactions are not well known. In this study, a Y-tube olfactometer was used to determine whether black soldier fly eggs, larvae, puparia (without pupae inside), adult carcasses, or resulting frass of black soldier fly decrease attractiveness of adult house flies to an oviposition substrate (i.e., sugar and wheat bran diet, used as the control). Secondly, two symbiotic bacteria associated were isolated, identified, and tested for house fly attractiveness with a Y-tube olfactometer. Thirdly, volatile organic compounds produced by swine feed before and after black soldier fly, or house fly larval digestion were analyzed with gas chromatography-mass spectrometry (GC-MS). Results indicate feed mixed with black soldier fly larval frass, and symbiotic bacteria, Paenalcaligenes sp. associated with black soldier fly, were less attractive to house flies than the control. However, another symbiotic bacterium Lysinibacillus sp. associated with black soldier fly pupae was attractive. In total, 52 volatile organic compounds were identified from treatments previously described. Quantitative differences in volatile profiles across treatments potentially explain the mechanism regulating house fly substrate choices in the presence, or absence, of black soldier fly larvae.

Applications of food waste-derived black soldier fly larval frass as incorporated compost, side-dress fertilizer and frass-tea drench for soilless cultivation of leafy vegetables in biochar-based growing media

• Food waste-derived black soldier fly larval frass and waste-wood biochar could substitute for peat compost and soil. • Sidedress application of frass yielded results comparable to those of the liquid inorganic fertilizer control. • The direct application of frass was more effective in promoting plant growth than frass tea application. • Frass and frass tea applications were free of heavy metals and foodborne pathogens. Black soldier fly (BSF) larval bioconversion can recycle nutrients in organic wastes into larval biomass and frass. While the frass has been commonly marketed as a soil amendment, its usefulness in soilless cultivation remains largely unexplored. Growth experiments were conducted to investigate the effectiveness of surplus food-derived and okara-derived BSF larval frass as an incorporated compost, side-dress fertilizer and frass-tea drench for the cultivation of pak choi and lettuce in waste-wood derived biochar growing media. Pak choi yields from treatments with surplus food-derived frass and biochar at a 10:90 (v/v) ratio and inorganic fertilizer were comparable to those of the control which consisted of soil, peat-based compost and inorganic fertilizer. However, yields decreased with increasing frass incorporation rates owing to high salinity and potentially low oxygen conditions in the growing media. When used as a fertilizer on biochar-coir growing media, the direct application of frass as a side-dress fertilizer was 1.6–6.8 times more effective in promoting lettuce growth than the application as a frass-tea drench. Frass fertilizers derived from surplus food outperformed those derived from okara by 1.3–5.3 times. Lettuce yields were not significantly different between the treatment with surplus food-derived frass applied as a side-dress fertilizer and the control of liquid inorganic fertilizer. Variations in fertilizing potential were attributed to nutrient availability and the presence of plant growth promoting microbes in the growing media. BSF larval frass derived from food waste shows promise in partially replacing unsustainable agricultural inputs for leafy vegetable cultivation, including soil and inorganic fertilizers.

Concentration and speciation of arsenic in an insect feeding on the leaves of Pteris vittata

The development of an effective and green bioinsecticide is a research hotspot. This study demonstrated the possibility of using an arsenic (As) hyperaccumulator as a bioinsecticide. When the As concentration in the Pteris vittata fronds exceeded 138 mg kg−1, the larva of the hawk moth (Theretra clotho) displayed apparent preference to lower-As-concentration P. vittata fronds. The As concentration in the larva body was as high as 850 mg kg−1. Such high concentration of As in the larva body might have been the case that T. clotho lacks a process to exclude As. The larval frass showed an As concentration of only 1%–4% of that in the larva body. The predominant As species in the larva body and frass was As(III)-SH. The percentage of As(III)-SH was slightly higher in the frass than that in the larval body. Chelation with thiols may be a universal detoxification mechanism for As in both plants and insects. In general, the adoption of P. vittata as a bioinsecticide should be feasible. However, the exact processes to achieve this goal still need further study. The mechanism of different animals to detoxify As is another interesting research topic.

Gut Structure and Microbial Communities in Sirex noctilio (Hymenoptera: Siricidae) and Their Predicted Contribution to Larval Nutrition.

The European woodwasp, Sirex noctilio Fabricius, is a major invasive quarantine pest that attacks and kills pine trees outside of its native range. Insect gut structure and gut microbiota play crucial roles in various life activities. Despite a few reports in nutrition and survival, an extensive study on the S. noctilio larval gut microbiome is lacking. We studied the gut structure using a stereo microscope and used high throughput sequencing of the bacterial 16S rRNA genes and fungal internal transcribed spacer 2 (ITS2) regions to investigate gut microbiota in different developmental stages of S. noctilio, including larvae, adults, and larval frass. We used PICRUSt2 to predict the functional profiles. The larval gut was thin and thread-like from the oral cavity to the anus, carrying few xylem particles in the crop. Pseudomonas, Ralstonia, and Burkholderia s.l were the dominant bacteria in the guts of larvae, adults, and frass, respectively. Even though Pseudomonas was the most abundant among all bacteria, Zoogloea, Ruminobacter, and Nitrosospira, which might be involved in degrading organic matter and fixing nitrogen occurred exclusively in the larval gut indicating their possible role in the growth and development of larvae in pine tree xylem. Fungal communities did not change significantly across different developmental stages or the frass. Amylostereum was dominant in the woodwasp’s larval gut. Functional prediction of bacterial and fungal communities revealed that they may encod enzymes involved in degrading lignocellulose and fixing nitrogen. Ours is the first study that compares gut microbial communities present in S. noctilio larvae, adults, and frass. This study could provide an understanding of larval nutrient acquisition in nutrient-deficient host xylem to some extent. Our study may unlock novel strategies for the development of pest management approaches based on interfering with the gut microbiota and restricting their role in larval survival and development.

Cottonseed Press Cake as a Potential Diet for Industrially Farmed Black Soldier Fly Larvae Triggers Adaptations of Their Bacterial and Fungal Gut Microbiota.

Black soldier fly larvae (Hermetia illucens, Diptera: Stratiomyidae) are used for the bioconversion of organic side products into valuable compounds such as proteins, lipids and chitin. However, the economic competitiveness of farmed insects compared to conventional protein production systems in agriculture and aquaculture depends on the availability of large quantities of inexpensive insect feed. Cottonseed press cake (CPC) is a side-stream of cotton production that is rich in proteins and lipids but unsuitable as feed for several farmed animals, except ruminants, due to the presence of the anti-nutritional sesquiterpenoid gossypol. Here, we tested CPC as a feed for black soldier fly larvae and studied the impact of this diet on the gut microbiome. Larvae reared on CPC developed normally and even showed a shorter life-cycle, but were smaller at the end of larval development than control larvae reared on chicken feed. The adaptability of the larvae to different diets is mediated by their versatile gut microbiome, which facilitates digestion and detoxification. We therefore used amplicon sequencing to analyze the bacterial and fungal communities associated with larvae reared on each diet, revealing differences between the larval guts and frass (residual feed substrate) as well as differences between the two diet groups. For example, Actinomycetaceae and Aspergillaceae were significantly enriched in guts of the CPC diet group and may help to metabolize compounds such as gossypol. Potentially probiotic yeasts and beneficial Enterobacteriaceae, which presumably belong to the core microbiota, were detected in high relative abundance in the gut and frass, indicating a functional role of these microbes, especially the protection against pathogens. We conclude that CPC may be suitable as an inexpensive and environmentally sustainable feed for the industrial rearing of black soldier flies.

Larval response to frass and guaiacol: detection of an attractant produced by bacteria from Spodoptera littoralis frass

Larval frass in herbivorous lepidopterans is mainly composed of plant-derived material and microbes from the gut. Despite the fact that frass from conspecific larvae repels female moths in Spodoptera littoralis from oviposition, the role of frass volatiles on larval foraging behavior is largely unknown. Here, we show that larvae of S. littoralis walk upwind to larval frass volatiles in a wind tunnel assay. We identified the frass volatile guaiacol (2-methoxyphenol) as key ligand for the S. littoralis odorant receptor (OR) SlitOr59 which we expressed heterologously. We isolated guaiacol-producing bacteria identified as Serratia marcescens from frass of larvae that were fed on cotton, and Enterobacter cloacae, E. ludwigii and Klebsiella sp. from frass derived from cabbage-fed larvae. In addition to guaiacol, we also identified volatiles acetoin, 3-methyl-1-butanol and dimethyl disulfide, in large proportions in headspace collections from the bacteria. A Y-tube olfactometer assay showed that fourth instar S. littoralis larvae are attracted to guaiacol. Moreover, cotton leaves treated with the insecticide Spinosad and guaiacol were highly attractive to the larvae. Our results provide a basis for management of the pest by directly targeting larvae, based on an attract-and-kill strategy. Further studies are needed to test the application of guaiacol for semiochemical-based pest management of Spodoptera pest species.

Evaluating the potential of okara-derived black soldier fly larval frass as a soil amendment

Bioconversion of organic waste by the black soldier fly (BSF) larva yields a by-product commonly known as ‘frass’. Although BSF larval frass has often been marketed as a biofertilizer, few studies have evaluated this claim. In this study, BSF larvae reared on a pure okara diet achieved an 85% waste reduction in the fresh weight of the okara. Subsequently, the frass was mixed with soil at concentrations of 10, 20, and 30% (vol/vol), and used to cultivate lettuce plants. At 10% concentration, the lettuce plants had biomasses comparable to those of the controls. Higher frass concentrations stunted the growth of the lettuce, likely because of the low C:N ratio of larval frass resulting from the rapid mineralization of nutrients. Larval frass was also found to be able to provide sufficient nutrients for lettuce growth as fertilizer application was only necessary after the first growth cycle, suggesting its suitability as a soil amendment. Analyses of the microbial community of all the growing media showed that the growth medium treatments with BSF larval frass tended to have a lower number of microbial species than the controls. Inherently higher micronutrient levels present in the frass resulted in the growth of lettuce plants. More importantly, the microbial analysis revealed that common foodborne pathogens were absent in the BSF larval frass and elemental analysis also indicated no heavy metal pollutants present. Overall, BSF larval frass was found to be a suitable soil amendment and more in-depth studies could facilitate its sensible use in agriculture.

Development of a loop-mediated isothermal amplification (LAMP) assay for the identification of the invasive wood borer Aromia bungii (Coleoptera: Cerambycidae) from frass

The red-necked longhorn beetle Aromia bungii (Faldermann, 1835) (Coleoptera: Cerambycidae) is native to east Asia, where it is a major pest of cultivated and ornamental species of the genus Prunus. Morphological or molecular discrimination of adults or larval specimens is required to identify this invasive wood borer. However, recovering larval stages of the pest from trunks and branches causes extensive damage to plants and is timewasting. An alternative approach consists in applying non-invasive molecular diagnostic tools to biological traces (i.e., fecal pellets, frass). In this way, infestations in host plants can be detected without destructive methods. This paper presents a protocol based on both real-time and visual loop-mediated isothermal amplification (LAMP), using DNA of A. bungii extracted from fecal particles in larval frass. Laboratory validations demonstrated the robustness of the protocols adopted and their reliability was confirmed performing an inter-lab blind panel. The LAMP assay and the qPCR SYBR Green method using the F3/B3 LAMP external primers were equally sensitive, and both were more sensitive than the conventional PCR (sensitivity > 103 to the same starting matrix). The visual LAMP protocol, due to the relatively easy performance of the method, could be a useful tool to apply in rapid monitoring of A. bungii and in the management of its outbreaks.

Low-cost technology for recycling agro-industrial waste into nutrient-rich organic fertilizer using black soldier fly

Efforts to recycle organic waste using black soldier fly (BSF) larvae into high-quality alternative protein ingredients in animal feeds and organic fertilizers have gained momentum worldwide. However, there is limited information on waste manipulation to increase nutrient retention for enhanced larval performance and frass fertilizer quality. In the present study, brewer’s spent grain with a carbon to nitrogen (C/N) ratio of 11 (control) was amended with sawdust to obtain substrates with C/N ratios of 15, 20, 25 and 30. The effects of substrate C/N ratios on BSF larval yield, waste degradation, biomass conversion efficiency, compost maturity and nutrient levels of frass fertilizer were evaluated. Substrates amended with sawdust did not significantly affect waste degradation efficiency and biomass conversion rates of BSF larvae. The wet and dried larval yields were significantly higher for substrates with C/N ratio of 15 compared to the other amended substrates. An amended substrate with C/N ratio of 15 enhanced nutrients uptake by BSF larvae, and increased nitrogen (N) and phosphorus retention in frass compost by 21 and 15%, respectively. Compost maturation time was shortened to five weeks, as indicated by the stable C/N ratios and high seed germination indices. This study has demonstrated that the amendment of the substrate with sawdust to C/N ratio of 15 could generate compost with desirable nutrients for use as high-quality fertilizer for organic farming.

Preventative Approach to Microbial Control of Capnodis tenebrionis by Soil Application of Metarhizium brunneum and Beauveria bassiana.

Management of the Mediterranean flat-headed root-borer, Capnodis tenebrionis, is critical due to the larvae’s root localization. Neonate larvae can be exposed to natural enemies before penetrating the roots. Application of Metarhizium brunneum strain Mb7 and Beauveria bassiana strain GHA formulations on rice granules was investigated for their efficacy against C. tenebrionis larvae. Mb7 application, evaluated on apricot twigs, significantly and dose-dependently reduced colonization rates of neonates, with highest mortality at 108 conidia/g soil. Neonate susceptibility to Mb7 and GHA was evaluated on potted rootstocks (GF677 almond × peach, 2729 plum) planted in entomopathogenic fungi (EPF)-premixed soil (1.3–1.6 × 105 conidia/cm3 soil) or in EPF-free soil surface-treated with 5 g Mb7 fungal granules (1.25 × 109 conidia). Larval colonization rates were reduced 7.4-fold in 2729 by both fungi; only Mb7 completely prevented colonization of GF677 by larvae. Larvae inside plant galleries exhibited mycosis with EPF-treated soils and both fungi proliferated on larval frass. Mb7 conidia germinated in the rhizosphere of GF677, and conidia of both fungi remained viable throughout the trial. Galleria baiting technique was used on EPF-treated soil to evaluate EPF infectivity over time; Mb7 and GHA persisted 180 and 90 days post inoculation, respectively. The formulation (fungus-covered rice grains), delivery method (mixing with soil) and persistence (3–6 months) of Mb7 and GHA are feasible for potential field application to control C. tenebrionis.

Symbiotic yeasts from the mycangium, larval gut and woody substrate of an African stag beetle Xiphodontus antilope (Coleoptera: Lucanidae).

Female stag beetles (Lucanidae) possess internal mycangia to maintain microbial cultures. Yeasts from these mycangia may help with larval nutrition in nutrient poor woody substrates, but only a few Lucanidae taxa have been studied and all reports originate from Europe and Asia. We identify the first mycangial yeasts of a South African endemic Lucanidae beetle, Xiphodontus antilope, using nuclear ribosomal RNA and ITS DNA sequence data. In addition we identified yeasts from the larval gut, fecal matter, frass and woody substrate surrounding larvae and pupae. The mycangium of X. antilope was confined to females and is structurally similar to all other Lucanidae. Unlike most Lucanidae that seemingly associate with single species of yeast, or whose mycangia contain yeast monocultures, three yeast species were commonly isolated from X. antilope. Scheffersomyces coipomoensis was the most numerically dominant species on most substrates and in most individuals, but a second, undescribed, Scheffersomyces species was present in high numbers. A third species, also undescribed and unrelated to Scheffersomyces, was recovered from all mycangia but could not be detected in the larval gut, fecal matter, frass or woody substrates. We confirm a close association of Scheffersomyces yeasts with Lucanidae globally, but other taxa may also be involved. We show that the predominant mycangial yeasts also form the predominant yeasts within the larval gut and the woody substrates around the larvae and pupae. This combined external and internal colonization by the same yeasts may provide enhanced opportunities for nutrient acquisition, but this needs validation in future studies.

Hermetia illucens (L.) larvae as chicken manure management tool for circular economy

The increased request for poultry meat and eggs of a rising human population requires more efficient and cleaner methods to manage increasing quantities of chicken manure. The black soldier fly Hermetia illucens is known as an efficient bio-converter of organic waste in proteins and fats, with the advantage that the larval frass is supposed to have compost-like properties. In the view to identify the operating conditions for the sustainable management and valorization of livestock waste at a pre-industrial scale, this study is aimed at: i) optimizing the growth of H. illucens on a mixture of chicken manure, chabazite and water; ii) assessing the soil amendment properties of the larval frass obtained from the optimized mixture. Preliminary trials allowed defining the basic rearing conditions in terms of temperature and substrate components. A mixture design based on a special cubic model allowed identifying the best mixture for H. illucens larvae growth, which consists in 34.5% chicken manure, 58.3% water and 7.2% coarse chabazite. This mix led to about 86% of alive prepupae weighting 90 mg on average, and to a reduction of the initial substrate amount by more than 75%. The larval frass obtained from this mixture showed soil improver properties, suggesting its use to supply the common peat based growing media for potted baby-leaf lettuce production. Overall, H. illucens larvae have proved to be a useful tool to favor a more sustainable management of chicken manure by strongly reducing its amount and closing its recovery cycle obtaining high value products for agricultural purposes.

Inspection of frass ejection could decrease the risk of white-spotted longicorn beetle Anoplophora malasiaca (Coleoptera: Cerambycidae) infestation of Japanese pine bonsais to negligible levels

To evaluate the infection risk of Anoplophora malasiaca (Thomson) (Coleoptera: Cerambycidae) in two species of Japanese pine bonsais (Japanese black pine and Japanese white pine), feeding and oviposition preferences of female A. malasiaca to pine bonsai and mandarin orange were examined. A female was released into a net cage together with a pair of one pine bonsai and one mandarin orange. All mandarin oranges used in preference tests were damaged by feeding and deep biting scars from females, and larval frass ejection from the trunk was found in some mandarin oranges within 4 months of observation. However, these scars were found on only a few pine bonsais, and frass ejection was not found in any pine bonsais. No A. malasiaca larvae were detected in these pine bonsais. Therefore, frass ejection from trunks might be indicative of the presence of larvae. In field surveys at bonsai nurseries, no A. malasiaca adults were found on the pine bonsais and no pine bonsais exhibited frass ejection. Statistical analyses of these results suggested that the risk of A. malasiaca infestation in pine bonsais could be ruled out as a negligible level by confirming an absence from the trunk of frass ejection.

Nutritional value of frass from black soldier fly larvae, Hermetia illucens, in a channel catfish, Ictalurus punctatus, diet

Frass is the by-product of the larval meal industry and includes larval waste, exoskeleton sheds and residual feed ingredients. Experimental frass was derived from the larvae of black solder flies fed distillers' dried grains with solubles and had a protein and fat content of 216 and 60 g/kg, respectively. A 10-week study was conducted to evaluate the effect of dietary levels of frass on growth, feed utilization, and body proximate and mineral composition of channel catfish, Ictalurus punctatus. Five diets containing 0, 50, 100, 200 and 300 g frass per kg diet were fed to channel catfish (5.24 ± 0.04 g) in quadruplicate aquaria to apparent satiation twice daily. Final weight gain was significantly increased in fish fed diets containing frass at levels from 100 to 300 g/kg. Fish fed diets without frass, and with 300 g/kg frass, showed the lowest and highest feed intake, respectively. Feed and protein efficiencies, however, were significantly lower in fish fed frass at levels of 200 g/kg and higher compared to the control diet. Survival, whole-body composition and mineral content were not affected by frass. In summary, black soldier fly larval frass has potential as a protein source or just an ingredient for enhancing palatability of catfish diets.