An Improved Chromosome-Level Genome Assembly of the Firefly Pyrocoelia pectoralis.

The endemic and endangered Chinese firefly Pyrocoelia pectoralis is a sexually dimorphic, nocturnal species. A previous attempt by this team to assemble a draft genome of P. pectoralis using PacBio and Illumina HiSeq X Ten platforms was limited in its usefulness by high redundancy and contamination. This prompted us to conduct an improved chromosome-level genome assembly of P. pectoralis. Ten chromosomes were further assembled based on Hi-C data to a 532.25 Mb final size with a 52.87 Mb scaffold N50. The total repeat lengths in the genome of P. pectoralis amount to 227.69 Mb; 42.78%. In total, 12,789 genes could be functionally annotated using at least one public database. Phylogenetic inference indicated that P. pectoralis and P. pyralis diverged ~51.41 million years ago. Gene family expansion and contraction analysis of 12 species were performed, and 546 expanded and 2660 contracted gene families were identified in P. pectoralis. We generated a high-quality draft of the P. pectoralis genome. This genome assembly should help promote research on the species' sexual dimorphism and its unique courtship behavior, which involves a combination of pheromonal and bioluminescent signals. It also can serve as a resource for accelerating genome-assisted improvements in the conservation of this species.

Sperm Competition and Paternity in the Endangered Firefly Pyrocoelia pectoralis (Coleoptera: Lampyridae: Lampyrinae).

The endangered terrestrial firefly Pyrocoelia pectoralis (Olivier) is endemic to China. Populations of P. pectoralis have decreased dramatically due to urbanization and pollution. Breeding and re-introduction to a suitable habitat may save the species from becoming extinct. Because of its polyandrous character, an investigation into the possibility of sperm competition and paternity outcomes from multiple matings was initiated to better understand its reproductive physiology. To achieve these goals, 13 SSR markers were developed. The results of paternity experiments indicate there is a significant difference between P3 and P1 or P2. The female reproductive system has three spermathecae which accept sperm from different matings, and no bursa or spermatophore-digesting organ is developed. Our research established that multiple inseminations with sperm from different males occur, leading to competition between ejaculates. The benefits of such competition include an increasing number of sperm in the ejaculates of competing males and the consequential increase in fertilized eggs (thus, fecundity), and thereby a higher chance of genetic diversity and fitness in the offspring of the firefly P. pectoralis.

A histochemical and morphological study of the mucus producing pedal gland system in Latia neritoides (Mollusca; Gastropoda; Hygrophila).

The freshwater gastropod Latia neritoides is endemic to the streams of New Zealand's North Island. This species has evolved a unique defence system: it exudes a luminescent mucus thought to deter predators. While the bioluminescence itself has been investigated before, the underlying gland system has remained unstudied and relevant information to understand the defence system has been missing till now. For the release of the glowing mucus of L. neritoides two places of origin were assumed: the lateral foot area or the mantel cavity. In this study the focus was on the first suggestion. To gain insight into the defence system, morphological as well as histochemical analyses were performed involving all secretory gland types in the sub-epithelial foot layer. The results were compared with the foot gland system of Neritina sp., a snail living in a comparable habitat, but using a different survival strategy. The gland types of the two gastropods were compared and their mucus types were investigated. Seven subepithelial gland cell types can be distinguished in the foot region of L. neritoides. Neritina sp., in contrast, has six gland cell types of which three laterally located ones are epithelial. Both species show a pedal gland in the anterior foot region. A striking difference between the species are two prominent subepithelial gland cell types (L1l/L2l) in the lateral foot area of L. neritoides, which are missing in Neritina sp. These gland cells are distributed throughout the entire lateral foot area of L. neritoides and make up about 85% of the mucus gland cells in this area. Defence mucus and trail mucus of L. neritoides show different specificities in lectin staining, but are not equally represented in the gland cell types. Yet, based on the huge size and high density of L1l and L2L, we envision a role for these gland types in the defence system.

Embracing Tradition: The Vital Role of Traditional Foods in Achieving Nutrition Security.

In a press release from the Convention on Biological Diversity (CBD) in 2019, it was reported that, currently, a mere 12 plants and 5 animal species account for approximately 75% of global food production [...].

Can Molecularly Engineered Plant Galls Help to Ease the Problem of World Food Shortage (and Our Dependence on Pollinating Insects)?

The world faces numerous problems and two of them are global food shortages and the dwindling number of pollinating insects. Plant products that do not arise from pollination are plant galls, which as in the case of oak apples, can resemble fruits and be the size of a cherry. It is suggested that once research has understood how chemical signals from gall-inducing insects program a plant to produce a gall, it should be possible to mimic and to improve nature and "bioengineer" designer galls of different sizes, colorations and specific contents to serve as food or a source of medicinally useful compounds. To achieve this objective, the genes involved in the formation of the galls need to be identified by RNA-sequencing and confirmed by gene expression analyses and gene slicing. Ultimately the relevant genes need to be transferred to naïve plants, possibly with the aid of plasmids or viruses as practiced in crop productivity increases. There is then even the prospect of engineered plant galls to be produced by plant tissue culture via genetic manipulation without the involvement of insects altogether.

An anatomical and ultrastructural study of the eye of the luminescent millipede Paraspirobolus lucifugus (Gervais 1836) (Diplopoda, Spirobolida, Spiroboleliidae).

The two forward-looking eyes and their ultrastructural organization of an 18 mm long adult bioluminescent female millipede (Paraspirobolus lucifugus) were investigated by transmission electron microscopy and energy-dispersive X-ray spectroscopy. Each eye contained approximately 23 ommatidia with 50-60 µm wide and 80 um thick corneal lenses that contained calcium and silicon and proximally ended in truncated flat surfaces of around 20 µm in diameter. A maximally 28 µm thick and 25 µm long rhabdom, made up of at least 12-14 retinula cells and a 4 µm thick sleeve of screening pigment granules in a light-adapted position was present. Compared with the eyes of non-luminescent julid millipede species, those of P. lucifugus share their basic anatomy, but also exhibit features like the wide possible binocular frontal visual overlap, somewhat narrower interommatidial angles combined with relatively larger rhabdoms, which suggests that P. lucifugus has more efficient eyes and makes greater use of its photoreceptors. P. lucifugus is negatively phototactic and strictly nocturnal and its activity rhythm is apparently governed by a circadian clock.

Interest in Insects as Food and Feed: It Does Not Wane in the Public Domain.

This Special Issue of Foods represents Volume 2 of the topic "Edible Insects as Innovative Foods: Nutritional, Functional and Acceptability Assessments" [...].

In Vitro Study of Cricket Chitosan's Potential as a Prebiotic and a Promoter of Probiotic Microorganisms to Control Pathogenic Bacteria in the Human Gut.

In this study, cricket chitosan was used as a prebiotic. Lactobacillus fermentum, Lactobacillus acidophilus, and Bifidobacterium adolescentis were identified as probiotic bacteria. Cricket chitin was deacetylated to chitosan and added to either De Man Rogosa and Sharpe or Salmonella/Shigella bacterial growth media at the rates of 1%, 5%, 10%, or 20% to obtain chitosan-supplemented media. The growth of the probiotic bacteria was monitored on chitosan-supplemented media after 6, 12, 24, and 48 h upon incubation at 37 °C. Growth of Salmonella typhi in the presence of probiotic bacteria in chitosan-supplemented media was evaluated under similar conditions to those of the growth of probiotic bacteria by measuring growth inhibition zones (in mm) around the bacterial colonies. All chitosan concentrations significantly increased the populations of probiotic bacteria and decreased the populations of pathogenic bacteria. During growth, there was a significant pH change in the media with all probiotic bacteria. Inhibition zones from probiotic bacteria growth supernatant against Salmonella typhi were most apparent at 16 mm and statistically significant in connection with a 10% chitosan concentration. This study suggests cricket-derived chitosan can function as a prebiotic, with an ability to eliminate pathogenic bacteria in the presence of probiotic bacteria.

Regeneration in Reptiles Generally and the New Zealand Tuatara in Particular as a Model to Analyse Organ Regrowth in Amniotes: A Review.

The ability to repair injuries among reptiles, i.e., ectothermic amniotes, is similar to that of mammals with some noteworthy exceptions. While large wounds in turtles and crocodilians are repaired through scarring, the reparative capacity involving the tail derives from a combined process of wound healing and somatic growth, the latter being continuous in reptiles. When the tail is injured in juvenile crocodilians, turtles and tortoises as well as the tuatara (Rhynchocephalia: Sphenodon punctatus, Gray 1842), the wound is repaired in these reptiles and some muscle and connective tissue and large amounts of cartilage are regenerated during normal growth. This process, here indicated as "regengrow", can take years to produce tails with similar lengths of the originals and results in only apparently regenerated replacements. These new tails contain a cartilaginous axis and very small (turtle and crocodilians) to substantial (e.g., in tuatara) muscle mass, while most of the tail is formed by an irregular dense connective tissue containing numerous fat cells and sparse nerves. Tail regengrow in the tuatara is a long process that initially resembles that of lizards (the latter being part of the sister group Squamata within the Lepidosauria) with the formation of an axial ependymal tube isolated within a cartilaginous cylinder and surrounded by an irregular fat-rich connective tissue, some muscle bundles, and neogenic scales. Cell proliferation is active in the apical regenerative blastema, but much reduced cell proliferation continues in older regenerated tails, where it occurs mostly in the axial cartilage and scale epidermis of the new tail, but less commonly in the regenerated spinal cord, muscles, and connective tissues. The higher tissue regeneration of Sphenodon and other lepidosaurians provides useful information for attempts to improve organ regeneration in endothermic amniotes.

Nutritional Composition of Honey Bee Drones of Two Subspecies Relative to Their Pupal Developmental Stages.

We examined the contents of nutritional importance, i.e., amino acids, fatty acids and minerals of different developmental stages of drones of two honey bee subspecies, namely Apis mellifera carnica and A. m. mellifera. The results revealed that, in general, individual amino acid amounts and therefore the total protein increased along with the developmental stages of the drones. No statistically significant differences were found between the same developmental stages of the two subspecies. The reverse, i.e., a decrease with developmental stage occurred in relation to the fatty acid composition. Most of the minerals were higher at advanced developmental stages. Overall, the high protein content (31.4-43.4%), small amount of fat (9.5-11.5%) and abundance of minerals such asiron and zinc, make drones a suitable nutritional resource. Even though nutrient content, especially protein, was higher in the pupae than the prepupae, we propose prepupae also as a commercial product based on their higher biomass production. Provided standard production protocols maintaining hygiene and safety will be adhered to, we propose that drone honey bees can be utilized as human food or animal feed.

Chemical Composition, Nutrient Quality and Acceptability of Edible Insects Are Affected by Species, Developmental Stage, Gender, Diet, and Processing Method.

Edible insects have been considered as either nutritious food itemsper se, or as wholesome ingredients to various dishes and components of traditional subsistence. Protein, fat, mineral and vitamin contents in insects generally satisfy the requirements of healthy food, although there is considerable variation associated with insect species, collection site, processing method, insect life stage, rearing technology and insect feed. A comparison of available data(based on dry weight) showed that processing can improve the nutrient content, taste, flavour, appearance and palatability of insects, but that there are additional factors, which can impact the content and composition of insect species that have been recommended for consumption by humans. This review focuses on factors that have received little attention in connection with the task to improve acceptability or choice of edible insects and suggests ways to guarantee food security in countries where deficiencies in protein and minerals are an acute and perpetual problem. This review is meant to assist the food industry to select the most suitable species as well as processing methods for insect-based food products.

Selection and Validation of Suitable Reference Genes for RT-qPCR Analysis in the Rare Aquatic Firefly Aquatica leii (Coleoptera: Lampyridae).

Aquatica leii Fu and Ballantyne is a species of rare aquatic firefly and endemic in China. It is considered good material to study the molecular mechanism of sexual flash communication systems. To improve conservation and behavioral research strategies, large-scale genetic studies involving gene-expression analysis are required and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is the most commonly used method. However, there have been very few reports on appropriate reference genes in any species of firefly. Here, we evaluated eight widely utilized reference genes including 18S, Actin, Reep5, Odc1, Tub, Gapdh, Ef1a and S27Ae for their expression stabilities in A. leii under three different conditions, i.e., life stage, tissue and dsRNA injection. Based on the gene stability ranking calculated by RefFinder, which integrates four algorithms (geNorm, delta Ct method, NormFinder, and BestKeeper), we recommend S27Ae and Reep5 as the most appropriate reference genes for molecular studies in different life stages; Ef1a and Odc1 for different tissues; Tub and Odc1 for RNAi studies. The most appropriate reference genes in all treatments are S27Ae and Tub. The results of this study will help improve accuracy and reliability to normalize RT-qPCR data in A. leii for further molecular analysis.

Observations on How People in Two Locations of the Plateau Département of Southeast Benin Perceive Entomophagy: A Study From West Africa.

We surveyed the local populations of Kétou and Pobè in Southeast Benin through interviews and with the aid of a semi-structured questionnaire in order to understand how they currently perceive entomophagy, an age-old tradition in their communities. The study revealed that the majority of the population was familiar with the use of insects as food, and a sizable number of people were still interested in insect consumption. Gender differences were not apparent. Tradition or culture was identified as the most influential factor, followed by taste, as determinants for eating or rejecting insects. However, identifying the edible species and comparing practices how they were prepared for consumption, we found that the knowledge was not homogenous across the society of Benin, with differences depending on ethnicity, culture, respondent's age, and educational background. Awareness and promotion of food insects in the society should help to preserve the practice of entomophagy and in turn could lead to the provision of much needed nutritional supplements to the poorer and disadvantaged sections of the society.

Chemical Composition and Nutritional Value of Different Species of Vespa Hornets.

We genetically identified three different species of hornets and analyzed the nutrient compositions of their edible brood. Samples were collected from a commercial production unit in Shizong province of China and from forests near Andong City in Korea. The species were identified as Vespa velutina, V. mandarinia, and V. basalis from China and V. velutina from Korea. Farmed V. velutina and V. mandarinia were found to have similar protein contents, i.e., total amino acids, whereas V. basalis contained less protein. The V. velutina brood collected from the forest contained the highest amount of amino acids. Altogether 17 proteinogenic amino acids were detected and quantified with similar patterns of distribution in all three species: leucine followed by tyrosine and lysine being predominant among the essential and glutamic acid among the non-essential amino acids. A different pattern was found for fatty acids: The polyunsaturated fatty acid proportion was highest in V. mandarinia and V. basalis, but saturated fatty acids dominated in the case of V. velutina from two different sources. The high amounts of unsaturated fatty acids in the lipids of the hornets could be expected to exhibit nutritional benefits, including reducing cardiovascular disorders and inflammations. High minerals contents, especially micro minerals such as iron, zinc, and a high K/Na ratio in hornets could help mitigate mineral deficiencies among those of the population with inadequate nutrition.

Entomotherapy: a study of medicinal insects of seven ethnic groups in Nagaland, North-East India.

BACKGROUND: The ethnic communities in Nagaland have kept a close relationship with nature since time immemorial and have traditionally used different kinds of insects and their products as folk medicine to treat a variety of human ills and diseases. The present study was conducted to record the entomotherapeutic practices of seven different ethnic groups of Nagaland. METHOD: Documentation is based on semi-structured questionnaires and group discussions with a total of 370 informants. The data collected were analysed using fidelity level (FL) and informant consensus factor (ICF). RESULTS: Fifty species of medicinal insects belonging to 28 families and 11 orders were identified in connection with treatments of at least 50 human ailments, of which the most frequently cited were coughs, gastritis, rheumatoid arthritis, stomach ache and wound healing. Mylabris sp. showed the highest fidelity level (FL) of 100% for its therapeutic property as a dermatologic agent, while the informant consensus factor (ICF) ranged from 0.66 to 1.00. The use of medicinal insects varies amongst the seven ethnic groups, suggesting that differences in cultures and geographic location can lead to the selection of specific insect species for specific medicinal purposes. The largest number of insect species appear to be used for treating gastrointestinal, dermatological and respiratory diseases. CONCLUSION: The list of medicinal insect species, many of which are reported for the first time in the present study, suggests the presence of a considerable diversity of therapeutically important insect species in the region and elaborate folk medicinal knowledge of the local ethnic groups. This knowledge of insects not just as a food, but also as therapy is passed down verbally from generation to generation, but is in danger of being lost if not documented in a systematic way. Having stood the test of time, traditional folk medicinal knowledge and its contribution through entomotherapy should not be regarded as useless as it has the potential to lead to the development of novel drugs and treatment methods.

Synodic lunar phases and suicide: based on 2605 suicides over 23 years, a full moon peak is apparent in premenopausal women from northern Finland.

Suicide data for this study were available for the period of March 1988 to June 2011, and involved 2111 male and 494 female victims from the Finnish province of Oulu. Data for lunar phases during that period were categorised into three groups: new moon (<25% visible), full moon (>75% visible) and other times with values in between. Seasonal effects were controlled with definitions for winter (Nov, Dec, Jan), spring (Feb, Mar, Apr), summer (May, June, July), and autumn (Aug, Sep, Oct). Suicide occurrences during different lunar phases were compared with their expected distribution using multinomial tests with all tests being two-tailed. Statistical significance was set at p < 0.05. No correlation between suicides and moon phase in any of the four seasons was apparent for male victims, but in winter for women it was (p = 0.001). Further analysis of the data revealed that the full moon association was statistically significant only for premenopausal women, defined as female victims younger than 45 years of age. To explain this unexpected finding a number of factors were considered, e.g., the darkness of a northern Finnish winter with increases of SAD and depression especially in premenopausal women, the influence of the lunar periodicity on the menstrual cycle, and cosmogeophysical effects on the humoral and autonomous nervous system.

Insects Used as Food and Feed: Isn't That What We All Need?

This Special Issue of Foods explores different aspects of how insects can be used as a novel resource for food and feed. Some contributions deal with questions of acceptability and legality, others tackle problems related to innovative techniques in processing and marketing food, and yet another group of papers highlights the use of insects and their bio-active products in the context of promoting human health. The collective aim of the contributions by the researchers from at least 20 countries is to examine whether the use of insects-be it for food, feed, or therapeutic purposes-has a future. We conclude that positive aspects undoubtedly exist regarding the nutritional and pharmacological value of various insect species but that environmental and bio-functional issues could even outweigh the nutritional value of food insects.

Squid sucker teeth and cocoons of a terrestrial flatworm: amino acid content of two nano-structurally identical tissues in phylogenetically unrelated taxa.

An extraordinary micro-structural similarity between squid sucker teeth and the egg shell of a terrestrial planarian worm has been reported, but to date only the amino acid content of the squid sucker tooth has been available. This prompted us to analyse the amino acid content of the planarian egg shell. Although both share an absence of detectable chitin and metal ions and both possess relatively high amounts of the amino acids GLY and HIS, the planarian egg shell is considerably richer in GLU, LYS and ASP. Most dramatic was the difference in TYR, which was the second most abundant amino acid in the squid, but hardly featured at all in the planarian egg shell. In the light of these new findings the different functional roles that the structures in question play in the lives of the two taxa are discussed. An EDS-analysis revealed clear C, N, and O peaks and additional very small peaks less than 0.1w% suggesting the presence of one or both S and Cl.