Tracking lipid synthesis using 2H2O and 2H-NMR spectroscopy in black soldier fly (Hermetia illucens) larvae fed with macroalgae.

Black soldier fly (Hermetia illucens) larvae are used to upcycle biowaste into insect biomass for animal feed. Previous research on black soldier fly has explored the assimilation of dietary fatty acids (FAs), but endogenous FA synthesis and modification remain comparatively unexplored. This study presents a 1H/2H-NMR methodology for measuring lipid synthesis in black soldier fly larvae using diluted deuterated water (2H2O) as a stable isotopic tracer delivered through the feeding media. This approach was validated by measuring 2H incorporation into the larvae's body water and consequent labelling of FA esterified into triacylglycerols. A 5% 2H enrichment in the body water, adequate to label the FA, is achieved after 24 h in a substrate with 10% 2H2O. A standard feeding trial using an invasive macroalgae was designed to test this method, revealing de novo lipogenesis was lower in larvae fed with macroalgae, probably related to the poor nutritional value of the diet.

Fatty Acid Profile and Escherichia coli and Salmonella sp. Load of Wild-Caught Seaweed Fly Fucellia maritima (Haliday, 1838) (Diptera: Anthomyiidae).

World aquaculture is expected to continue to grow over the next few decades, which amplifies the need for a higher production of sustainable feed ingredients for aquatic animals. Insects are considered good candidates for aquafeed ingredients because of their ability to convert food waste into highly nutritional biomass. However, commercially available terrestrial insect species lack n-3 long-chain polyunsaturated fatty acids (LC-PUFAs), which are essential biomolecules for marine cultured species. Nevertheless, several coastal insect species feature LC-PUFAs in their natural fatty acid (FA) profile. Here, we analysed the lipidic profile of wild-caught seaweed fly Fucellia maritima, with a focus on their FA profile, to evaluate its potential to be used as an aquafeed ingredient, as well as to screen for the presence of pathogenic bacteria. Results showed that the flies had a total lipid content of 13.2% of their total dry weight. The main classes of phospholipids (PLs) recorded were phosphatidylethanolamines (PEs) (60.8%), followed by phosphatidylcholine (PC) (17.1%). The most abundant FA was palmitoleic acid (C16:0) with 34.9% ± 4.3 of total FAs, followed by oleic acid (C18:1) with 30.4% ± 2.3. The FA composition of the flies included essential fatty acids (EFAs) for both freshwater fish, namely linoleic acid (C18:2 n-6) with 3.4% ± 1.3 and alpha-linoleic acid (C18:3 n-3) with 3.4% ± 1.9, and marine fish, namely arachidonic acid (C20:4 n-6) with 1.1% ± 0.3 and eicosapentaenoic acid (C20:5 n-3) with 6.1% ± 1.2. The microbiological analysis found 9.1 colony-forming units per gram (CFU/g) of Enterobacteriaceae and no presence of Salmonella sp. was detected in a sample of 25 g of fresh weight. These findings indicate that Fucellia maritima biomass holds the potential to be used as an additional aquafeed ingredient due to its FA profile and the low count of pathogenic bacteria, which can contribute to the optimal growth of fish and shrimp with a low risk of pathogen transfer during the feed production chain.

Body size-dependent effects on the distribution patterns of phoretic mite species assemblages on Rhynchophorus ferrugineus (Olivier, 1790).

Phoretic mites attach to different body parts of the red palm weevil (RPW), Rhynchophorus ferrugineus (Olivier, 1790), to disperse. However, the question of how the patterns of attachment sites are formed remains intriguing. Here, we examined RPW-associated phoretic mites in four districts in Northern Portugal (macrohabitat), and investigated the patterns of mite distribution on six body parts of RPW (microhabitat). At the macrohabitat level, we detected seven phoretic mite taxa using the RPW host in each of the four studied districts, all documented for the first time in association with this invasive exotic species in Portugal. However, their relative abundance (species evenness) varied between districts, as did species diversity. All examined weevils carried mites, and the prevalence of the different taxa did not differ between districts or sex of weevils. Measured by mean abundance and degree of aggregation, Centrouropoda sp. proved to be the dominant taxon, while Acarus sp. and Curculanoetus rhynchophorus were considered common subordinate taxa and Uroovobella sp., Mesostigmata, Nenteria extremica and Dendrolaelaps sp. sparse taxa. At the microhabitat level, all taxa were present on all body parts of the RPW; the highest abundance was in a region encompassing the inner surface of the elytra and the membranous hind wings (subelytral space). Analysis of niche overlap revealed that the distribution patterns of phoretic mite taxa on the RPW were not randomly structured. In the subelytral space, interspecific coexistence of mites increased as a function of body size difference with the dominant Centrouropoda sp. We found that in the subelytral space the large dominant species Centrouropoda sp. displaced the larger species Uroobovella sp. and the similarly sized species Nenteria extremica, but coexisted with smaller taxa.

Bioconversion of olive oil pomace by black soldier fly increases eco-efficiency in solid waste stream reduction producing tailored value-added insect meals.

Olive oil is one of the most important agricultural products in Mediterranean areas, and currently the European Union is the largest producer. Due to technological innovations, Portugal has become one of the main olive oil producing countries over the last few years, accompanied by large amounts of olive oil pomace (OOP), the most representative residue of the olive oil extraction process. This is causing serious waste management problems since current management solutions also present environmental impacts. Here we explored the black soldier fly (Hermetia illucens) potential to biotransform OOP into valuable insect meals by feeding them OOP-based diets as substrates. Results show that despite survival rates not being affected by higher replacement (75% and 50%) levels of OOP, there was an increase in larval instar duration. Substrate reduction was significantly lower for higher replacement levels but was not affected up to the 50% replacement level. Feed conversion rate differed among all the treatments, increasing as the replacement level increased, while bioconversion rate, which also differed among all the treatments, decreased as replacement level increased. Differences in larval protein content were only seen at higher replacement levels (75%), with an increase in protein content for replacements of up to 25%. One of the most striking results was the change in fatty acid profile, which became more abundant in monounsaturated fatty acids (mostly oleic acid) as the olive pomace replacement levels increased in comparison with the control substrate, rich in saturated fatty acids (palmitic acid). These results show that BSF can be an effective OOP bioconversion agent, and resulting insect meals can be used as alternatives to currently available saturated fatty acid insect meals.

A roadmap for ladybird conservation and recovery.

Ladybirds (Coleoptera: Coccinellidae) provide services that are critical to food production, and they fulfill an ecological role as a food source for predators. The richness, abundance, and distribution of ladybirds, however, are compromised by many anthropogenic threats. Meanwhile, a lack of knowledge of the conservation status of most species and the factors driving their population dynamics hinders the development and implementation of conservation strategies for ladybirds. We conducted a review of the literature on the ecology, diversity, and conservation of ladybirds to identify their key ecological threats. Ladybird populations are most affected by climate factors, landscape composition, and biological invasions. We suggest mitigating actions for ladybird conservation and recovery. Short-term actions include citizen science programs and education, protective measures for habitat recovery and threatened species, prevention of the introduction of non-native species, and the maintenance and restoration of natural areas and landscape heterogeneity. Mid-term actions involve the analysis of data from monitoring programs and insect collections to disentangle the effect of different threats to ladybird populations, understand habitat use by taxa on which there is limited knowledge, and quantify temporal trends of abundance, diversity, and biomass along a management-intensity gradient. Long-term actions include the development of a worldwide monitoring program based on standardized sampling to fill data gaps, increase explanatory power, streamline analyses, and facilitate global collaborations.

Las catarinas (Coleoptera: Coccinellidae) proporcionan servicios que son críticos para la producción de alimento, y juegan un papel ecológico como fuente de alimento para depredadores. Sin embargo, la riqueza, abundancia y distribución de catarinas están en peligro debido a muchas amenazas antropogénicas. La carencia de conocimiento sobre el estatus de conservación de la mayoría de las especies y los factores que inciden en su dinámica poblacional dificulta el desarrollo e implementación de estrategias de conservación para las catarinas. Realizamos una revisión de la literatura sobre la ecología, diversidad y conservación de catarinas para identificar sus amenazas ecológicas clave. Las poblaciones de catarinas fueron afectadas mayormente por factores climáticos, composición del paisaje e invasiones biológicas. Proponemos acciones de mitigación para la conservación y recuperación de catarinas. Acciones a corto plazo incluyen programas de ciencia y educación ciudadana, medidas de protección para la recuperación de hábitat y de especies amenazadas, prevención de la introducción de especies no nativas y el mantenimiento y restauración de áreas naturales y la heterogeneidad del paisaje. Acciones a mediano plazo implican el análisis de datos obtenidos de programas de monitoreo y colecciones de insectos para desenmarañar el efecto de las diferentes amenazas a las poblaciones de catarinas, comprender el uso del hábitat por taxa de los que se tiene conocimiento limitado y cuantifica las tendencias temporales de la abundancia, diversidad y biomasa a lo largo de un gradiente de intensidad de manejo. Acciones a largo plazo incluyen el desarrollo de un programa de monitoreo a nivel mundial basado en muestreos estandarizados para subsanar la falta de datos, incrementar el poder explicativo, optimizar los análisis y facilitar colaboraciones globales.

The Influence of Non-Optimal Rearing Conditions and Substrates on the Performance of the Black Soldier Fly (Hermetia illucens).

Among the insect species reared as alternative protein sources, Hermetia illucens (black soldier Fly, BSF) has shown a huge potential mostly due to its high protein content, its bioconversion rates, and versatility in using different feeding substrates. Insect rearing may use continuous or batch feeding regimes and, among the used substrates, supermarket feedstock waste has gained recent interest under a circular economy perspective, but several uncertainties remain regarding the heterogeneity and the potential effects of the quantity and quality of these substrates on BSF larvae (BSFL) development. In this experimental work, five replicates of a hundred BSFL were fed in a continuous feeding regime, using seven different isolated vegetables as substrates (wheat bran, pumpkin, apple, grape pomace, red onion, red cabbage, and spinach), at three different temperatures (20, 25, and 30 °C) and two substrate moisture conditions (natural and 70% substrate moisture), until 50% of the larvae achieved the prepupal stage. BSFL performance and bioconversion parameters were evaluated. Our results show that some substrates should be avoided when rearing Hermetia illucens on feedstocks. Among these, apple feed led to poorer and slower development performances with more than 100 days of larval stage, while grape pomace and spinach showed higher mortality rates, which may be due to some anti-nutritional compounds. Larvae fed on pumpkin, red cabbage, and red onion presented good bioconversion results with higher values of efficiency of conversion of digested feed between 14.4 and 25. This work delivers relevant results for black soldier fly reared on a continuous feeding system using vegetable feedstock substrates and their potential trade-offs.

Gall midge Baldratia salicorniae Kieffer (Diptera: Cecidomyiidae) infestation on Salicornia europaea L. induces the production of specialized metabolites with biotechnological potential.

Saltmarsh plants have several defense mechanisms against threatening abiotic conditions, such as salinity, inundation, or exposure to intense radiation, less is known regarding response to insect pests attack. Salicornia europaea L. plant stands are produced as cash crops in Portuguese coastal areas. In 2017, these crops suffered significant attacks from a gall midge fly (Baldratia salicorniae Kieffer), reducing its economic value. To understand how this attack influenced S. europaea chemical composition, infested and non-infested branches were collected, and their extracts were analysed by GS-MS and UHPLC-MS. Results revealed that different degrees of infestations displayed different chemical composition. Several compounds were for the first time identified in S. europaea, such as, arachidic acid, alpha-tocopherol, henicos-1-ene, and squalene. Most evident results were the reduced amount of alkanes in the infested conditions, which seems to be a direct consequence of insect infestation. Several compounds identified in the infested branches are known to have negative effects on insect larvae by reducing larval growth (linoleic acid) or increasing insect mortality (oleic acid). Halophyte plants production is increasing and it is accompanied by the urge to develop early control strategies against potential pests. These strategies may include ecological friendly solutions such as endogenous production of specialized metabolites to retrieve plant self-defences. Further, our results showed that B. salicorniae herbivory also induced the production of higher number of specialized metabolites with important known biological activities. In years in which high infestations reduce organoleptic qualities for fresh consumption plants can be used in biorefinery industries for metabolite extraction.

Bioconversion and performance of Black Soldier Fly (Hermetia illucens) in the recovery of nutrients from expired fish feeds.

In modern aquaculture systems, feed is the main source of the waste being produced, including expired aquafeeds. There is a link between the expiration date of aquafeeds enriched with fish oil for marine fish and the observation of several physical and microbiological changes. Among these, lipid oxidation is worth highlighting, as this process is responsible for the loss of palatability of aquafeeds, which can lead to feeding rejection by the species being farmed. In this study, we used an expired fish aquafeed, which otherwise would be discarded as waste, as a substrate to feed Black Soldier Fly (BSF) larvae. Different replacement levels of expired aquafeed were used which unravelled the amount of n-3 fatty acids added to larval tissues of BSF larvae after 2, 7, and 10 days of feeding. Our results also showed that shorter trials and higher diet replacement levels induced a deleterious effect on final larval weight. Furthermore, amino acid and fatty acid larval contents were shaped by the supplied diet, with results supporting the inclusion of BSF meal in aquafeeds, due to the levels of lysine (5.6-8.9%), methionine (1.9-3.2%), and omega-3 fatty acids (14.5%) recorded. These results demonstrate that the re-introduction of an expired resource aiming to diversify the source of aquafeeds raw materials can be safely achieved through BSF biotransformation. Overall, BSF larvae can successfully recover important nutrients for aquafeeds targeting marine species and foster the production of value-added insects under a circular bioeconomy framework.

An annotated checklist of ladybeetle species (Coleoptera, Coccinellidae) of Portugal, including the Azores and Madeira Archipelagos.

A comprehensive annotated checklist of the ladybeetle species of Portugal, including the Azores and Madeira archipelagos, is presented. The Coccinellidae fauna comprises a total of 101 species: 83 from the Mainland, 39 from Madeira, and 32 from the Azores. The listed species are distributed among 2 sub-families and 13 tribes: within the subfamily Microweiseinae, Madeirodulini (1 species), Serangiini (2 species), and within the subfamily Coccinellinae, Azyini (1 species), Chilocorini (4 species), Coccidulini (7 species), Coccinellini (30 species), Epilachnini (4 species), Hyperaspidini (7 species), Noviini (2 species), Platynaspini (1 species), Scymnini (37 species), Stethorini (3 species), and Sticholotidini (2 species). The Portuguese fauna comprises 10 exotic species: 5 present in the Mainland, 7 in Madeira, and 6 in the Azores. Harmoniaaxyridis (Pallas, 1773) from Madeira, Propyleaquatuordecimpunctata (Linnaeus, 1758) from the Azores, Delphastuscatalinae (Horn, 1895) from the Azores and Madeira, Nephus (Geminosipho) reunioni (Fürsch, 1974) and Nephus (Nephus) voeltzkowi Weise, 1910 from Madeira and Microserangium sp. from the Mainland, are reported for the first time. Some species are considered doubtful records, as explained in the text. These results were obtained by compiling information on the available literature regarding ladybeetle species on the Portuguese mainland and insular territories, and original data.

Nonlinearities lead to qualitative differences in population dynamics of predator-prey systems.

Since typically there are many predators feeding on most herbivores in natural communities, understanding multiple predator effects is critical for both community and applied ecology. Experiments of multiple predator effects on prey populations are extremely demanding, as the number of treatments and the amount of labour associated with these experiments increases exponentially with the number of species in question. Therefore, researchers tend to vary only presence/absence of the species and use only one (supposedly realistic) combination of their numbers in experiments. However, nonlinearities in density dependence, functional responses, interactions between natural enemies etc. are typical for such systems, and nonlinear models of population dynamics generally predict qualitatively different results, if initial absolute densities of the species studied differ, even if their relative densities are maintained. Therefore, testing combinations of natural enemies without varying their densities may not be sufficient. Here we test this prediction experimentally. We show that the population dynamics of a system consisting of 2 natural enemies (aphid predator Adalia bipunctata (L.), and aphid parasitoid, Aphidius colemani Viereck) and their shared prey (peach aphid, Myzus persicae Sulzer) are strongly affected by the absolute initial densities of the species in question. Even if their relative densities are kept constant, the natural enemy species or combination thereof that most effectively suppresses the prey may depend on the absolute initial densities used in the experiment. Future empirical studies of multiple predator - one prey interactions should therefore use a two-dimensional array of initial densities of the studied species. Varying only combinations of natural enemies without varying their densities is not sufficient and can lead to misleading results.