The pederin-producing bacteria density dynamics in Paederus fuscipes at different developmental stages.

Pederin, a defensive toxin in Paederus fuscipes, is produced by an uncultured Gram-negative symbiont, which establishes a stable symbiotic relationship with a female host before completion of metamorphosis. However, the transmission process of pederin-producing bacteria (PPB) in P. fuscipes at different life stages remains unknown. Herein, the PPB population dynamics and transcriptome atlas for P. fuscipes development (egg, first-instar larva, second-instar larva, pupa, and newly emerged female and male) were characterised. We found that a microbial layer containing PPB covered the eggshell, which could be sterilised by smearing the eggshell with streptomycin. Maternal secretions over the eggshell are likely the main PPB acquisition route for P. fuscipes offspring. The PPB density in eggs was significantly higher than that in other life stages (p < 0.05), which demonstrated that the beetle mothers gave more PPB than the larvae acquired. Physiological changes (hatching and eclosion) led to a decreased PPB density in P. fuscipes. Pattern recognition receptors related to Gram-negative bacteria recognition were identified from P. fuscipes transcriptomes across various life stages, which might be used to screen genes involved in PPB regulation. These results will help advance future efforts to determine the molecular mechanisms of PPB colonisation of P. fuscipes.

Differences in microbiome composition and transcriptome profiles between male and female Paederus fuscipes harbouring pederin-producing bacteria.

Pederin, a group of antitumor compounds, is produced by an endosymbiotic bacterium of Paederus fuscipes. Pederin content differed between male and female P. fuscipes, but the reason why these differences are maintained remains unexplored. Here, the pederin-producing bacteria (PPB) infection rate in P. fuscipes was investigated. Furthermore, we assessed the microbiota structure differences in male and female P. fuscipes harbouring PPB and sequenced the transcriptome of both sexes to shed light on genes of interest. Of the 625 analysed beetles (275 females, 350 males), 96.36% of females and 31.14% of males were positive for PPB infection. PPB accounted for 54.36%-82.70% of the bacterial population in females but showed a much lower abundance in males (0.92%-3.87%). Reproductive organs possessed the highest PPB abundance compared with other parts of females, but no such relationships existed in males. Moreover, we provide the first transcriptome analysis of male and female P. fuscipes harbouring PPB and identified 8893 differentially expressed unigenes. Our results indicated that the pederin content difference between males and females might be caused by the PPB density difference in hosts. The biosequence data would be helpful for illustrating the mechanism that regulates PPB density in P. fuscipes.

Natural products as drugs and tools for influencing core processes of eukaryotic mRNA translation.

Eukaryotic protein synthesis is the highly conserved, complex mechanism of translating genetic information into proteins. Although this process is essential for cellular homoeostasis, dysregulations are associated with cellular malfunctions and diseases including cancer and diabetes. In the challenging and ongoing search for adequate treatment possibilities, natural products represent excellent research tools and drug leads for new interactions with the translational machinery and for influencing mRNA translation. In this review, bacterial-, marine- and plant-derived natural compounds that interact with different steps of mRNA translation, comprising ribosomal assembly, translation initiation and elongation, are highlighted. Thereby, the exact binding and interacting partners are unveiled in order to accurately understand the mode of action of each natural product. The pharmacological relevance of these compounds is furthermore assessed by evaluating the observed biological activities in the light of translational inhibition and by enlightening potential obstacles and undesired side-effects, e.g. in clinical trials. As many of the natural products presented here possess the potential to serve as drug leads for synthetic derivatives, structural motifs, which are indispensable for both mode of action and biological activities, are discussed. Evaluating the natural products emphasises the strong diversity of their points of attack. Especially the fact that selected binding partners can be set in direct relation to different diseases emphasises the indispensability of natural products in the field of drug development. Discovery of new, unique and unusual interacting partners again renders them promising tools for future research in the field of eukaryotic mRNA translation.

Potassium permanganate use in the management of a wound induced by contact with Paederus: A case report.

Pederin is a vesicant toxic amide, found in the hemolymph of the beetle genus, Paederus. Physical contact with these insects produces a type of irritant dermatitis with variable clinical symptoms. It has been identified that Pederin (a vesicant toxic amide responsible for the ulcerative lesion) is produced by Gram-negative endosymbiotic bacteria of the Pseudomonas genus. The present study describes the case of a patient who had come into contact with the insect Paederus sp. and developed an ulcerative lesion. He was first treated with topical steroids, as part of the conventional management, and following no response to treatment, he was treated locally with a 5% potassium permanganate solution, in conjunction with systemic antibiotic therapy, obtaining a good response in the healing process. On the whole, the present study demonstrates that potassium permanganate solutions, in conjunction with antibiotics and anti-inflammatories, may be beneficial in the treatment of dermatitis or ulcerative lesions caused by insects of the Paederus genus. However, further research is required to fully determine the complete benefits and any side-effects.

The structure and diversity of microbial communities in Paederus fuscipes (Coleoptera: Staphylinidae): from ecological paradigm to pathobiome.

BACKGROUND: Paederus fuscipes is medically the most famous rove beetle, which causes dermatitis or conjunctivitis in humans, as well as gastrointestinal toxicosis in livestock, via releasing toxic hemolymph containing pederin. Pedrin biosynthesis genes have been identified in uncultured Pseudomonas-like endosymbionts that are speculated to be acquired through a horizontal transfer. However, the composition of the P. fuscipes microbial community, especially of the gut and genital microbiome, remains unclear. This study was aimed to characterize the structure and diversity of P. fuscipes-associated bacterial communities in terms of gender, organ, and location using the Illumina HiSeq platform in the southern littorals of Caspian Sea. RESULTS: The OTUs identified from P. fuscipes specimens were collapsed into 40 phyla, 112 classes, 249 orders, 365 families, 576 genera, and 106 species. The most abundant families were Pseudomonadaceae, Spiroplasmataceae, Weeksellaceae, Enterococcaceae, and Rhizobiaceae, respectively. Thirty top genera made up > 94% of the P. fuscipes microbiome, with predominating Pseudomonas, followed by the Spiroplasma, Apibacter, Enterococcus, Dysgonomonas, Sebaldella, Ruminococcus, and Wolbachia. Interesting dissimilarities were also discovered within and between the beetle microbiomes in terms of genders and organs. Analyses showed that Spiroplasma / Apibacter as well as Pseudomonas / Pseudomonas were the most abundant in the genitals / intestines of male and female beetles, respectively. Bacterial richness did not display any significant difference in the three provinces but was higher in male beetles than in females and more in the genitals than intestines. CONCLUSIONS: The present study identified Pseudomonas-like endobacterium as a common symbiont of P. fuscipes beetles; this bacterium begins its journey from gut and genitalia of females to reach the male rove beetles. Additionally, male and female rove beetles were characterized by distinctive microbiota in different organs, likely reflecting different functions and/or adaptation processes. Evidence of the extension of P. fuscipes microbiome from the environmental paradigm to the pathobiome was also presented herein. A comprehensive survey of P. fuscipes microbiome components may eventually lead to ecological insights into the production and utilization of defensive compound of pederin and also the management of linear dermatitis with the use of available antibiotics against bacterial pathogens released by the beetles. Video Abstract.

Bacterial diversity and community structure shapes pederin polymorphism but lacks association with host genotype specificity in the rove beetle, Paederus fuscipes.

Paederus beetles are notorious for their irritant haemolymph toxin pederin, which is synthesised by an unculturable bacterial symbiont, causing dermatitis when in contact with human skin. Pederin polymorphism is observed when (+) females carrying the toxin pederin and (-) females lacking this co-occur in natural populations. Despite reports detecting pederin polymorphism in several Paederina beetles, symbiont infection frequencies in their natural populations and the bacterial diversity differences underlying the polymorphism between these female types are understudied. Herein we report a high prevalence (>80%) of female Paederus fuscipes carrying bacterial ped genes in all six study populations. This finding suggests that selection pressure favouring pederin-producing females is crucial for survival in the natural environment. 16S rRNA metabarcoding analysis revealed significant dominance of the unculturable pederin-producing Pseudomonas-like bacterium (Gammaproteobacteria) in (+) females, consistent with previous studies. The microbial diversity of the (-) females revealed a significant abundance of Apibacter (Bacteroidia), previously undocumented, suggesting its importance in the functionality of (-) females. Predicted functions related to metabolisms are enriched in (-) females, suggesting fitness advantage possibilities in sustaining the population in the absence of predation. Further investigations on the possible genetic basis of the host genotype revealed no association of host mtDNA and pederin polymorphism in Pa. fuscipes.

Diaphorin, a Polyketide Produced by a Bacterial Symbiont of the Asian Citrus Psyllid, Inhibits the Growth and Cell Division of Bacillus subtilis but Promotes the Growth and Metabolic Activity of Escherichia coli.

Diaphorin is a polyketide produced by "Candidatus Profftella armatura" (Gammaproteobacteria: Burkholderiales), an obligate symbiont of a notorious agricultural pest, the Asian citrus psyllid Diaphorina citri (Hemiptera: Psyllidae). Diaphorin belongs to the pederin family of bioactive agents found in various host-symbiont systems, including beetles, lichens, and sponges, harboring phylogenetically diverse bacterial producers. Previous studies showed that diaphorin, which is present in D. citri at concentrations of 2 to 20 mM, has inhibitory effects on various eukaryotes, including the natural enemies of D. citri. However, little is known about its effects on prokaryotic organisms. To address this issue, the present study assessed the biological activities of diaphorin on two model prokaryotes, Escherichia coli (Gammaproteobacteria: Enterobacterales) and Bacillus subtilis (Firmicutes: Bacilli). Their growth and morphological features were analyzed using spectrophotometry, optical microscopy followed by image analysis, and transmission electron microscopy. The metabolic activity of E. coli was further assessed using the ß-galactosidase assay. The results revealed that physiological concentrations of diaphorin inhibit the growth and cell division of B. subtilis but promote the growth and metabolic activity of E. coli. This finding implies that diaphorin functions as a defensive agent of the holobiont (host plus symbionts) against some bacterial lineages but is metabolically beneficial for others, which potentially include obligate symbionts of D. citri. IMPORTANCE Certain secondary metabolites, including antibiotics, evolve to mediate interactions among organisms. These molecules have distinct spectra for microorganisms and are often more effective against Gram-positive bacteria than Gram-negative ones. However, it is rare that a single molecule has completely opposite activities on distinct bacterial lineages. The present study revealed that a secondary metabolite synthesized by an organelle-like bacterial symbiont of psyllids inhibits the growth of Gram-positive Bacillus subtilis but promotes the growth of Gram-negative Escherichia coli. This finding not only provides insights into the evolution of microbiomes in animal hosts but also may potentially be exploited to promote the effectiveness of industrial material production by microorganisms.

Co-occurrence of pederin-producing and Wolbachia endobacteria in Paederus fuscipes Curtis, 1840 (Coleoptera: Staphilinidae) and its evolutionary consequences.

The dual occurrence of Pseudomonas-like and Wolbachia endobacteria has not been investigated in the Pederus beetles yet. We investigated pederin-producing bacteria (PPB) infection in Paederus fuscipes specimens from the southern margins of the Caspian Sea by designed genus-specific (OprF) and species-specific (16S rRNA) primers. Wolbachia infection was studied through a nested-PCR assay of Wolbachia surface protein (wsp) gene. Of the 125 analyzed beetles, 42 females (82.35%) and 15 males (20.27%) were positive to PPB infection; this is the first study reporting male P. fuscipes infection to PPB. Wolbachia infection was found in 45 female (88.23%) and 50 male (67.57%) analyzed beetles. Surprisingly, a number of 36 females (70.59%) and 13 males (17.57%) were found to be infected with both PPB and Wolbachia endosymbionts. In general, population infection rates to PPB and Wolbachia were determined to be 45.6% and 76%, respectively. The infection rates of female beetles to PPB and PPB-Wolbachia were significantly higher than males. In Paederus species, only female beetles shelter PPB and the discovery of this bacterium in adult males may reflect their cannibalistic behavior on the contaminated stages. Phylogenetic analysis showed that the sequences of OprF gene were unique among Pseudomonas spp.; however, sequences of 16S rRNA gene were related to the PPB of Pederus species. The co-occurrence and random distribution of these endobacteria may imply putative tripartite interactions among PPB, Wolbachia, and Paederus. In order to elucidate these possible tripartite interactions, further studies are required even at gender level.

Genome of Labrenzia sp. PHM005 Reveals a Complete and Active Trans-AT PKS Gene Cluster for the Biosynthesis of Labrenzin.

The complete genome of the strain Labrenzia sp. PHM005, a free-living producer of a pederin analog 18-O-demethyl pederin, hereinafter labrenzin, has been sequenced. This strain contains two replicons comprising a circular chromosome of 6,167,349 bp and a circular plasmid (named p1BIR) of 19,450 bp. A putative gene cluster responsible for the synthesis of labrenzin (lab cluster) has been identified showing that it encodes a trans-AT mixed type PKS/NRPS biosynthetic pathway that is responsible for the synthesis of pederin and possibly an onnamide analog. The putative boundaries of the lab gene cluster were determined by genetic comparisons with other related strains, suggesting that the cluster consists of a 79-kb region comprising 3 genes encoding multidomain hybrid polyketide synthase/non-ribosomal peptide synthetase (PKS/NRPS) proteins (PKS4, PKS/NRPS13, and PKS/NRPS15), and 16 auxiliary enzymes. Transcriptomic analyses suggest that all the genes of the cluster are expressed in our culture conditions (i.e., in minimal medium in the absence of any specific inducer) at detectable levels. We have developed genetic tools to facilitate the manipulation of this strain and the functional characterization of the cluster genes. We have created a site-directed mutant unable to produce pederin, demonstrating experimentally for the first time the role of the cluster in the synthesis of pederin. This work paves the way to unravel the clues of the biosynthesis of pederin family compounds and opens the door to modify and overproduce these anticancer drugs for industrial and pharmaceutical purposes.

Crystal structure of diaphorin methanol monosolvate isolated from Diaphorina citri Kuwayama, the insect vector of citrus greening disease.

The title compound C22H39NO9·CH3OH [systematic name: (S)-N-((S)-{(2S,4R,6R)-6-[(S)-2,3-di-hydroxy-prop-yl]-4-hy-droxy-5,5-di-methyl-tetra-hydro-2H-pyran-2-yl}(hy-droxy)meth-yl)-2-hy-droxy-2-[(2R,5R,6R)-2-meth-oxy-5,6-dimeth-yl-4-methyl-ene-tetra-hydro-2H-pyran-2-yl]acetamide methanol monosolvate], was isolated from the Asian citrus psyllid, Diaphorina citri Kuwayama, and crystallizes in the space group P21. 'Candidatus Profftella armatura' a bacterial endosymbiont of D. citri, biosynthesizes diaphorin, which is a hybrid polyketide-nonribosomal peptide comprising two highly substituted tetra-hydro-pyran rings joined by an N-acyl aminal bridge [Nakabachi et al. (2013 ▸). Curr. Biol.23, 1478-1484]. The crystal structure of the title compound establishes the complete relative configuration of diaphorin, which agrees at all nine chiral centers with the structure of the methanol monosolvate of the di-p-bromo-benzoate derivative of pederin, a biogenically related compound whose crystal structure was reported previously [Furusaki et al. (1968 ▸). Tetra-hedron Lett.9, 6301-6304]. Thus, the absolute configuration of diaphorin is proposed by analogy to that of pederin.

Dermatitis por Paederus. Acerca de tres casos / Paederus dermatitis. About three cases

Resumen La "dermatitis por Paederus", es una dermatitis vesicante secundaria al contacto con coleópteros de este género. Se describen más de 600 especies de Paederus, con predominio en áreas tropicales y subtropicales, varias de ellas se asocian a dermatitis. Al ser apretado o aplastado contra la piel, la hemolinfa que contiene paederina, lesiona la epidermis, por medio del bloqueo de la mitosis de células epiteliales basales y células suprabasales. Se exponen tres casos clínicos dermatológicos con lesiones características compatibles con "dermatitis por Paederus". Se realiza además una revisión bibliográfica sobre el tema, a fin de exponer los puntos más importantes de esta patología de gran interés médico y que representa en muchas ocasiones desafíos diagnósticos.

Summary "Paederus dermatitis" is a vesicant dermatitis secondary to contact with this beetle. There are more than 600 species of Paederus described, with a predominance in tropical and subtropical areas, some of these are associated with dermatitis. When they are pressed or crushed against the skin, releases secretions with pederin, that causes an injury in the epidermis, by mitosis blocking of basal and suprabasal epithelial cells. This article describes three dermatological clinical cases with charac-teristic lesions compatible with "Paederus dermatitis". In addition a bibliographic review is presented in order to expose the most important points of this medical interest pathology, that frequently represents diagnostic challenges.

Differential efficacy of toxic pederin in deterring potential arthropod predators of Paederus (Coleoptera: Staphylinidae) offspring.

This study investigates the effects of pederin, a hemolymph toxin that is accumulated in the eggs of most Paederus females, on potential arthropod predators of the offspring of P. fuscipes and P. riparius. Insects generally do not respond to pederin present in the prey. Paederus larvae are sufficiently agile to escape from these predators by running away, and the eggs are hidden by the females. Unlike insects, (wolf) spiders are deterred by prey with pederin. They turn away from larvae they have already captured and exhibit cleansing behavior. Larvae containing pederin survive the attacks of spiders without damage, whereas larvae descended from females that do not transfer pederin into their eggs are often killed and eaten. In the case of sudden attacks by spiders, the larvae have no chance of escape. Their survival thus depends on chemical defense. These investigations show for the first time why pederin might be of considerable importance for Paederus in the field.

Paederus fuscipes dermatitis: a report of nine cases observed in Italy and review of the literature.

Paederus sp. is a beetle belonging to Staphylinidae family and Coleoptera order. Its distribution is worldwide, especially in hot climates. Over 600 species of Paederus are known, approximately 50 are able to cause an irritant contact dermatitis. When the beetle is accidentally crushed on the skin, it releases pederin, a potent toxin with vesicating action. In Europe, only anecdotical cases of Paederus sp. dermatitis have been reported. Since 1993, we have observed approximately 25 patients with suspected Paederus sp. dermatitis. In 9 the clinical diagnosis was confirmed because Paederus fuscipes were found. The case list includes 6 males and 3 females, ages ranging from 6 to 53 years (mean age: 26.3 years). Six patients presented with one lesion and 3 with 2 lesions. Eyelids (3 patients), shoulders (3), neck (2), cheek (1), breast (1), back (1) and calf (1) were involved. All patients showed erythema, 2 oedema, 2 blisters, 1 vesicles, 1 pustules and 1 crusts. In the patient with pustules, bacteriological examinations were negative. A biopsy was carried out in 4 patients. In the early stages, spongiosis with exocytosis of neutrophils was observed. Vesicles with neutrophils and necrosis of the epidermis were typical of the advanced stage. Oedema of the dermis and a mixed infiltrate, with eosinophils, neutrophils and lymphocytes, in the papillary and upper reticular dermis, were also observed. Direct immunofluorescence was negative in all patients. Paederus sp. should also be considered in temperate climates as a possible aetiological agent in cases of acute dermatitis.

Rove beetles of medical importance in Brazil (Coleoptera, Staphylinidae, Paederinae)

Rove beetles of medical importance in Brazil (Coleoptera, Staphylinidae, Paederinae). The rove beetles of the genus Paederus Fabricius, 1775 are the most important group within Coleoptera causing dermatitis around the world. The medical importance of Paederus depends on its toxic hemolymph released when these beetles are crushed on human skin. The effects are mainly dermatitis linearis and some sporadic cases of conjunctivitis. In Brazil seven species of Paederus are known to cause dermatitis: P. amazonicus Sharp, 1876, P. brasiliensis Erichson, 1840, P. columbinus Laporte, 1835, P. ferus Erichson, 1840, P. mutans Sharp, 1876, P. protensus Sharp, 1876 stat. rev., and Paederus rutilicornis Erichson, 1840. Paederus mutans and P. protensus are for the first time recorded as of medical importance, whereas the record of P. rutilicornis in Brazil is doubtful. All seven species are redescribed and a dichotomous key is provided. The geographic distributions of all species are documented. The results provided here include the most recent and relevant taxonomic revision of Paederus of the Neotropical region, the first identification key for Brazilian species and the increase of recorded species of medical importance in the world.

Medically important beetles (insecta: coleoptera) of Iran

This study focused on coleopteran species that are responsible for the emergence of recent cases of dermatological manifestations in Iran. To the best of our knowledge, five species of the family Meloidae and nine species of the genus Paederus are by far the only beetles recognized as medically important in Iran. The staphylinids consists of Paederus ilsae, P. iliensis, P. fuscipes, P. kalalovae, P. balcanicus, P. lenkoranus, P. littoralis, P. carpathicus, P. nigricornis, while the meloids are Mylabris impressa, M. guerini, Muzimes iranicus, Alosimus smyrnensis and Epicauta sharpi. Most cases of linear dermatitis in this country occur in areas bordering the Caspian Sea. This problem is caused by beetles of the genus Paederus which are present as adults from mid-April to October with particularly high incidences from May to August. Fars (in southern Iran) ranks second in number of cases of insect-induced dermatitis. The third major region in which this type of dermatitis has been recorded is Hamedan Province, in the west of the country. Meloid dermatitis showed its highest severity in 2001, when a considerable number of patients sought medical help in Toyserkan and Nahavand counties. New cases of skin blistering were reported along the Persian Gulf coast and the agent was identified as Epicauta sharpi (Coleoptera: Meloidae). In all these regions, it was observed that recorded cases of lesions coincided precisely with the yearly peaks of the beetles. Paederus fuscipes and P. kalalovae are the predominant species along the Caspian Sea shore. It appears that P. fuscipes is homogeneously distributed throughout the Caspian Sea region while the distribution of the other species is more irregular. Paederus fuscipes is probably the major agent that causes linear dermatitis in northern Iran. Whereas this disease is a rural difficulty in the south, mainly in villages or small towns, it is an urban problem in northern provinces along the Caspian Sea shore. Meloid dermatitis has been registered only in western and southern Iran. It is not as widespread as linear dermatitis and remains a minor rural health problem.(AU)