Chryseobacterium oryctis sp. nov., isolated from the gut of the beetle Oryctes rhinoceros, and Chryseobacterium kimseyorum sp. nov., isolated from a stick insect rearing cage.

Two strains of Chryseobacterium identified from different experiments are proposed to represent new species. Strain WLa1L2M3T was isolated from the digestive tract of an Oryctes rhinoceros beetle larva. Strain 09-1422T was isolated from a cage housing the stick insect Eurycantha calcarata. Sequence analysis of the 16S rRNA and rpoB genes found both strains to be similar but not identical to other Chryseobacterium species. Whole-genome sequencing suggested the isolates represent new species, with average nucleotide identity values ranging from 74.6 to 80.5 %. Genome-to-genome distance calculations produced values below 25.3 %, and digital DNA-DNA hybridization values were 13.7-29.9 %, all suggesting they are distinct species. The genomic DNA G+C content of WLa1L2M3T is approximately 32.53 %, and of 09-1422T is approximately 35.89 %. The predominant cellular fatty acids of strain WLa1L2M3T are C15 : 0 iso, summed feature 9 (C16 : 0 10OH or C17 : 1 iso ω6c), C17 : 0 iso 3OH, summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), C15 : 0 iso 3OH, C15 : 0 anteiso and C13 : 0 iso, and those of strain 09-1422T are C15 : 0 iso, summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), C17 : 0 iso 3OH, C15 : 0 anteiso, C15 : 0 iso 3OH, C16 : 1 ω7c, C17 : 0 2OH and C18 : 0. In addition, physiological and biochemical tests revealed phenotypic differences from related Chryseobacterium type strains. These cumulative data indicate that the two strains represent novel species of the genus Chryseobacterium for which the names Chryseobacterium oryctis sp. nov. and Chryseobacterium kimseyorum sp. nov. are proposed with WLa1L2M3T (=BCRC 81350T=JCM 35215T=CIP 112035T) and 09-1422T (=UCDFST 09-1422T=BCRC 81359T=CIP 112165T), as type strains, respectively.

Thiamine (vitamin B1) as an insect repellent: a scoping review.

While the desire for systemic repellents is high, ineffective repellents put one at risk of insect-vectored pathogens. Vitamin B1, or thiamine, has been touted as a systemic insect repellent since 1943, and denounced as an ineffective placebo for just as long. This paper presents a scoping review of 104 relevant case reports, research studies, and review articles to trace the evolution of this idea and identify an evidence-based, scientific consensus. Reports of thiamine's systemic repellency are primarily anecdotal and based on uncontrolled trials and/or used bite symptoms as a proxy for reduced biting. Controlled experiments on insect landing and feeding found no evidence of repellency. Of the 49 relevant review papers, 16 insect bite prevention guidelines, and 4 government documents, none after the 1990s claimed thiamine is a repellent. The findings of this review are that thiamine cannot repel arthropods in any dosage or route of administration. Due to limited available evidence, the possibility that thiamine reduces the subjective symptoms of insect bites cannot currently be ruled out. Unfortunately, many medical professionals and travelers today still believe thiamine may be effective despite the evidence stating otherwise. Continued promotion of debunked repellents on the commercial market poses a serious risk in countries with the endemic, mosquito-vectored disease.

Pseudomonas schmalbachii sp. nov., isolated from the gut of a millipede (Trigoniulus corallinus) from a coconut tree.

During an investigation of microbes associated with arthropods living in decaying coconut trees, a Pseudomonas isolate, Milli4T, was cultured from the digestive tract of the common Asian millipede, Trigoniulus corallinus. Sequence analysis of 16S rRNA and rpoB genes found that Milli4T was closely related but not identical to Pseudomonas panipatensis Esp-1T, Pseudomonas knackmussi B13T and Pseudomonas humi CCA1T. Whole genome sequencing suggested that this isolate represents a new species, with average nucleotide identity (OrthoANIu) values of around 83.9-87.7% with its closest relatives. Genome-to-genome distance calculations between Milli4T and its closest relatives also suggested they are distinct species. The genomic DNA G+C content of Milli4T was approximately 65.0 mol%. Phenotypic and chemotaxonomic characterization and fatty acid methyl ester analysis was performed on Milli4T and its related type strains. Based on these data, the new species Pseudomonas schmalbachii sp. nov. is proposed, and the type strain is Milli4T (=BCRC 81294T=JCM 34414T=CIP 111980T).

Who's afraid of DEET? Fearmongering in papers on botanical repellents.

DEET (N,N-Diethyl-meta-toluamide) is considered the gold standard in mosquito repellents, not only for its effectiveness, but also for its safety. DEET has been more extensively studied for safety than any other repellent, and is accepted as completely safe when used correctly (i.e. not consumed or bathed in). Researchers studying botanical repellents, however, often paint DEET as far more toxic than it really is, falsely claiming it is a menace to the public health or even the environment. These claims are unfounded, and often the only evidence given by such publications are references to other publications also studying botanical repellents. Such publications are biased, and may be attacking DEET's excellent safety record to justify their existence and the need for their research. The inconvenient yet undisputable fact is that no botanical repellent has been proven to be as safe as DEET, and the majority never had any safety testing whatsoever. The automatic assumption that botanical repellents are safer than DEET is the 'appeal to nature fallacy,' which also drives most of the market for "natural" repellents, yet natural repellents have side effects and even a body count. Finding a botanical repellent that works as well as DEET and is equally safe is a legitimate research goal on its own, and need not be justified by fear-mongering and irrational chemophobia. Researchers studying these alternatives should strive for integrity, raising the real issue of the lack of safety testing for botanical repellents rather than denying the proven safety of DEET.

Transcriptome and microbiome of coconut rhinoceros beetle (Oryctes rhinoceros) larvae.

BACKGROUND: The coconut rhinoceros beetle, Oryctes rhinoceros, is a major pest of palm crops in tropical Asia and the Pacific Islands. Little molecular data exists for this pest, impeding our ability to develop effective countermeasures and deal with the species' growing resistance to viral biocontrols. We present the first molecular biology analyses of this species, including a metagenomic assay to understand the microbiome of different sections of its digestive tract, and a transcriptomics assay to complement the microbiome data and to shed light on genes of interest like plant cell wall degrading enzymes and immunity and xenobiotic resistance genes. RESULTS: The gut microbiota of Oryctes rhinoceros larvae is quite similar to that of the termite gut, as both species feed on decaying wood. We found the first evidence for endogenous beta-1,4-endoglucanase in the beetle, plus evidence for microbial cellobiase, suggesting the beetle can degrade cellulose together with its gut microfauna. A number of antimicrobial peptides are expressed, particularly by the fat body but also by the midgut and hindgut. CONCLUSIONS: This transcriptome provides a wealth of data about the species' defense against chemical and biological threats, has uncovered several potentially new species of microbial symbionts, and significantly expands our knowledge about this pest.

Differential expression of endogenous plant cell wall degrading enzyme genes in the stick insect (Phasmatodea) midgut.

BACKGROUND: Stick and leaf insects (Phasmatodea) are an exclusively leaf-feeding order of insects with no record of omnivory, unlike other "herbivorous" Polyneoptera. They represent an ideal system for investigating the adaptations necessary for obligate folivory, including plant cell wall degrading enzymes (PCWDEs). However, their physiology and internal anatomy is poorly understood, with limited genomic resources available. RESULTS: We de novo assembled transcriptomes for the anterior and posterior midguts of six diverse Phasmatodea species, with RNA-Seq on one exemplar species, Peruphasma schultei. The latter's assembly yielded >100,000 transcripts, with over 4000 transcripts uniquely or more highly expressed in specific midgut sections. Two to three dozen PCWDE encoding gene families, including cellulases and pectinases, were differentially expressed in the anterior midgut. These genes were also found in genomic DNA from phasmid brain tissue, suggesting endogenous production. Sequence alignments revealed catalytic sites on most PCWDE transcripts. While most phasmid PCWDE genes showed homology with those of other insects, the pectinases were homologous to bacterial genes. CONCLUSIONS: We identified a large and diverse PCWDE repertoire endogenous to the phasmids. If these expressed genes are translated into active enzymes, then phasmids can theoretically break plant cell walls into their monomer components independently of microbial symbionts. The differential gene expression between the two midgut sections provides the first molecular hints as to their function in living phasmids. Our work expands the resources available for industrial applications of animal-derived PCWDEs, and facilitates evolutionary analysis of lower Polyneopteran digestive enzymes, including the pectinases whose origin in Phasmatodea may have been a horizontal transfer event from bacteria.

Mitigation Strategies against Food Safety Contaminant Transmission from Black Soldier Fly Larva Bioconversion.

The black soldier fly larva, Hermetia illucens, can efficiently convert organic waste into biomatter for use in animal feed. This circularity comes with a risk of contaminating downstream consumers of the larval products with microbes, heavy metals, and other hazards potentially present in the initial substrate. This review examines research on mitigation techniques to manage these contaminants, from pretreatment of the substrate to post-treatment of the larvae. While much research has been done on such techniques, little of it focused on their effects on food safety contaminants. Cheap and low-technology heat treatment can reduce substrate and larval microbial load. Emptying the larval gut through starvation is understudied but promising. Black soldier fly larvae accumulate certain heavy metals like cadmium, and their ability to process certain hazards is unknown, which is why some government authorities are erring on the side of caution regarding how larval bioconversion can be used within feed production. Different substrates have different risks and some mitigation strategies may affect larval rearing performance and the final products negatively, so different producers will need to choose the right strategy for their system to balance cost-effectiveness with sustainability and safety.

Coconut rhinoceros beetle digestive symbiosis with potential plant cell wall degrading microbes.

Coconut rhinoceros beetle (CRB, Oryctes rhinoceros) is an invasive palm pest whose larvae eat wood, yet lack the necessary digestive enzymes. This study confirmed endogenous CRB cellulase is inactive, suggesting microbial fermentation. The inner lining of the CRB hindgut has tree-like structures covered with a conspicuous biofilm. To identify possible symbionts, 16 S rRNA amplicon sequencing was used on individuals from across Taiwan. Several taxa of Clostridia, an anaerobic class including many cellulolytic bacteria, were highly abundant in most individuals from all locations. Whole metagenome sequencing further confirmed many lignocellulose degrading enzymes are derived from these taxa. Analyses of eggs, larvae, adults, and soil found these cellulolytic microbes are not transmitted vertically or transstadially. The core microbiomes of the larval CRB are likely acquired and enriched from the environment with each molt, and enable efficient digestion of wood.

Mad scientist: the unique case of a published delusion.

In 1951, entomologist Jay Traver published in the Proceedings of the Entomological Society of Washington her personal experiences with a mite infestation of her scalp that resisted all treatment and was undetectable to anyone other than herself. Traver is recognized as having suffered from Delusory Parasitosis: her paper shows her to be a textbook case of the condition. The Traver paper is unique in the scientific literature in that its conclusions may be based on data that was unconsciously fabricated by the author's mind. The paper may merit retraction on the grounds of error or even scientific misconduct "by reason of insanity," but such a retraction raises the issue of discrimination against the mentally ill. This article asks what responsibilities journals have when faced with delusions disguised as science, what right editors have to question the sanity of an author, and what should be done about the Traver paper itself. By placing higher emphasis on article content than author identity, scientific integrity is maintained and a balance is struck between avoiding discrimination against the mentally ill and not preventing patients from seeking needed treatment.

A practical guide to collections-based research on ecogeographic rules.

Ecogeographic research into how species' forms vary across space, time, and climate has taken on new urgency due to contemporary global climate change. Research using museum specimens and other records to study biological rules like Bergmann's, Allen's, and Gloger's Rules has a long history and continues to generate publications and robust scientific debates. Despite the prevalence and history of the field, however, no simple guide on how to carry out such work has ever been published. To lower the barriers of entry for new researchers, this review was created as a practical guide on how to perform ecogeographic research. The guide consolidates disparately published methodologies into a single, convenient document that reviews the history and present of the field of ecogeographic rule research, and describes how to generate appropriate hypotheses, design experiments, gather, and analyze biotic and geographic data, and interpret the results in an ecologically meaningful manner. The result is a semi-standardized guide that enables scientists at all levels from any institution to carry out an investigation from start to finish on any biological rule, taxon, and location of their choice.

Anatomical changes of the beetle digestive tract during metamorphosis correspond to dietary changes.

During pupation, the tissues of holometabolous insects change in preparation for the adult lifestyles, although little literature exists examining this hidden process in detail. Using beetles as a model, we hypothesized that species where the adult and larva have the same diets will show less pronounced changes of the digestive tract during metamorphosis than species where the adults diets differ. We also wanted to observe these changes and document them at a level of detail missing from the current record. We compared the structure of the digestive tracts of scarab beetles Oryctes rhinoceros, Thaumastopeus shangaicus, and Protaetia spp. (Coleoptera: Scarabaeidae)-where the larvae eat wood, soil, or compost while the adults feed on soft plant matter, tree sap, and rotting fruits-with the tortoise beetle, Cassida circumdata (Coleoptera: Chrysomelidae), which feeds on leaves as both larva and adult. In the scarab beetles we observed considerable changes in the digestive tracts during the pupal stage, which we could divide into distinct stages, while in the leaf beetle pupae, the gut did not change. This information can provide new insight into metamorphosis, and the illustrations of what occurs during pupation are novel contributions to this field that will facilitate future work.

Where are we now? Bergmann's rule sensu lato in insects.

Bergmann's rule states that individuals of a species/clade at higher altitudes or latitudes will be larger than those at lower ones. A systemic review of the known literature on inter- and intraspecific variation in insect size along latitudinal or altitudinal clines was done to see how often such clines appeared and whether they reflected classwide, species-specific, or experimentally biased tendencies. Nearly even numbers of studies showed Bergmann clines and converse-Bergmann clines, where insects get smaller as latitude/altitude increases. In fact, the majority of studies suggested no clines at all. Small ranges may have obscured certain clines, while giant ranges may have introduced artifacts. Researchers examining interspecific patterns found clines less frequently than those examining intraspecific patterns because of variation among species within the clades, which renders interspecific studies unhelpful. Bergmann's rule does not apply to hexapods with nearly the same consistency as it does to endothermic vertebrates. The validity of Bergmann's rule for any group and range of insects is highly idiosyncratic and partially depends on the study design. I conclude that studies of Bergmann's rule should focus within species and look at widespread but contiguous populations to account for all sources of variation while minimizing error.

Cytochrome P450 Genes Expressed in Phasmatodea Midguts.

Cytochrome P450s (CYPs) are xenobiotic detoxification genes found in most eukaryotes, and linked in insects to the tolerance of plant secondary chemicals and insecticide resistance. The number and diversity of CYP clans, families, and subfamilies that an organism produces could correlate with its dietary breadth or specialization. This study examined the CYP diversity expressed in the midguts of six species of folivorous stick insects (Phasmatodea), to identify their CYP complement and see if any CYPs correlate with diet toxicity or specialization, and see what factors influenced their evolution in this insect order. CYP genes were mined from six published Phasmatodea transcriptomes and analyzed phylogenetically. The Phasmatodea CYP complement resembles that of other insects, though with relatively low numbers, and with significant expansions in the CYP clades 6J1, 6A13/14, 4C1, and 15A1. The CYP6 group is known to be the dominant CYP family in insects, but most insects have no more than one CYP15 gene, so the function of the multiple CYP15A1 genes in Phasmatodea is unknown, with neofunctionalization following gene duplication hypothesized. No correlation was found between CYPs and diet specialization or toxicity, with some CYP clades expanding within the Phasmatodea and others likely inherited from a common ancestor.

Critical commentary on "The potential of a site-specific delivery of thiamine hydrochloride as a novel insect repellent exerting long-term protection on human skin: In-vitro, ex-vivo study and clinical assessment".

This letter comments on a recent article by Halawani et al. (10.1016/j.xphs.2021.07.017), which claimed a complex hydrogel formulation of thiamine nanospheres is a topical insect repellent. The authors did not thoroughly review the extensive prior literature on the subject that found no evidence of repellency for thiamine, and the experiment described lacked negative controls. Its results are not conclusive.

Increasing Body Sizes in Anomala expansa expansa (Coleoptera: Scarabaeidae) Populations in Response to Rising Temperatures Over Time.

Many insects have been studied over wide geographical areas to determine whether they follow Bergmann's Rule, which predicts that animal clades or populations should have smaller body sizes in warmer climates. While this ecogeographic rule is well supported in mammals and birds, insect latitudinal sizes can show Bergmann, converse Bergmann, or no size clines at all. Museum collections are typical sources of data for insect clines, and long-term collections should reflect rising global temperatures and shifting climates, possibly producing temporal size clines along with any geographical clines. We hypothesize that insects with Bergmann clines geographically will show Bergmann-like clines temporally as well, and that the converse and inverse of this rule are also true. By looking at museum samples going back a century, we tested whether Anomala expansa expansa (Bates, 1866), a species of Scarabaeidae beetle common in lowland Taiwan, was experiencing long-term changes in body size in response to rising temperatures. We found that overall, the size of these beetles increased over time. Within Taipei populations, this increase was correlated with rising average yearly temperatures. The impact of this pest species' rising size with time will need to be monitored, and temporal size clines in other pests need to be investigated.

Syzygium samarangense Leaf Infusion as Aedes albopictus (Diptera: Culicidae) Ovitrap Bait.

Adding lures can improve the efficiency of mosquito ovitraps used for monitoring or in attract-and-kill pest management. Easily produced, low-to-no cost bait would be ideal for remote field sites and community-run vector management. Plant infusions are popular ovitrap baits for their low cost and potent attractiveness, attributed either to the plants or their microbiomes. We tested fermented leaf infusions of the wax apple tree, Syzygium samarangense (Blume) Merr. & Perry, as bait in lethal ovitraps in urban Taipei with Bti larvicide. All trapped insects were inferred to be Asian tiger mosquito, Aedes albopictus (Skuse). The bait significantly increased the yield of the traps, with no negative interactions with the larvicide in either direction. Syzgium samarangense leaves are readily available across Taiwan, making their infusion an easy bait to greatly improve ovitrap efficiency.

Evolution of CP2 transcription factors in Hexapoda.

The CP2 transcription factors are highly conserved in metazoans, where they are divided into two groups: grainyhead and late SV40 factor (LSF). We traced their evolutionary history in the Hexapoda using over 500 insect transcriptomes, to test the hypothesis that the evolution of holometaboly involved novel isoforms of these genes. All insects appear to express at least one grainyhead and one LSFlike gene, regardless of life cycle, as in most known metazoa. No major evolutionary events in these gene families occurred during the evolution of insects.

Detection of deformed wing virus (DWV) in the Vietnamese walking stick Medauroidea extradentata (Phasmatodea).

Deformed wing virus (DWV) is a single-stranded positive sense RNA virus that mainly infects honey bees (Apis mellifera) and can have devastating impacts on the colony. Recent studies have shown the presence of this virus in several species of Apis spp. and some other Hymenoptera, but our knowledge of their host range is very limited. We screened previously sequenced RNAseq libraries from different tissues of Vietnamese Walking Stick, Medauroidea extradentata (Phasmatodea) for DWV. We only found this virus in six libraries from anterior and posterior midgut tissue. From the midgut libraries we were able to construct a complete DWV genome sequence, which consisted of 10,140 nucleotides and included one open reading frame. Pairwise genome comparison confirmed strong similarity (98.89 %) of these assembled sequences with only 113 SNPs to the original DWV genome. We hypothesize the M. extradentata acquired this virus via a foodborne transmission by consuming DWV-infected material such as pollen or leaves contaminated with virus infected bee faeces.

Potential of Black Soldier Fly Production for Pacific Small Island Developing States.

Organic waste such as food waste and livestock manure is a serious concern in the Pacific Islands, where landfills are overflowing and illegal dumping of waste threatens the fragile ecosystems. Organic waste also attracts filth flies, some of which are vectors for pathogens that cause human disease. The black soldier fly, Hermetia illucens, has tremendous potential for the Pacific Islands. Capable of digesting almost any organic matter and converting it into insect biomass, black soldier flies are already being used around the world to process organic waste into larvae. The system can be adapted to large-scale municipal composting as well as small sizes for individual livestock farms or even urban households. The larvae can be fed live to fish or poultry, processed into feed comparable to fishmeal or soy meal, or even used to generate biofuel. Thus, the fly not only eliminates waste, but also can improve the sustainability of livestock production. The Pacific Small Island Developing States stand to benefit immensely from black soldier fly bioconversion facilities, used primarily as a means to compost organic waste; however, several knowledge gaps must first be addressed. We reviewed the state of black soldier flies in the Pacific and identified where their use shows the most promise. Research priorities for the field include fly surveys and bioconversion assays using Pacific crop waste.