Venom Alkaloids Research Articles

Biological Activities and Ecological Significance of Fire Ant Venom Alkaloids.

Venoms produced by arthropods act as chemical weapons to paralyze prey or deter competitors. The utilization of venom is an essential feature in the biology and ecology of venomous arthropods. Solenopsis fire ants (Hymenoptera: Formicidae) are medically important venomous ants. They have acquired different patterns of venom use to maximize their competitive advantages rendered by the venom when facing different challenges. The major components of fire ant venom are piperidine alkaloids, which have strong insecticidal and antibiotic activities. The alkaloids protect fire ants from pathogens over the course of their lives and can be used to defend them from predators and competitors. They are also utilized by some of the fire ants' natural enemies, such as phorid flies to locate host ants. Collectively, these ants' diverse alkaloid compositions and functions have ecological significance for their survival, successful invasion, and rapid range expansion. The venom alkaloids with powerful biological activities may have played an important role in shaping the assembly of communities in both native and introduced ranges.

Venom Peptides, Polyphenols and Alkaloids: Are They the Next Antidiabetics That Will Preserve β-Cell Mass and Function in Type 2 Diabetes?

Improvement of insulin secretion by pancreatic β-cells and preservation of their mass are the current challenges that future antidiabetic drugs should meet for achieving efficient and long-term glycemic control in patients with type 2 diabetes (T2D). The successful development of glucagon-like peptide 1 (GLP-1) analogues, derived from the saliva of a lizard from the Helodermatidae family, has provided the proof of concept that antidiabetic drugs directly targeting pancreatic β-cells can emerge from venomous animals. The literature reporting on the antidiabetic effects of medicinal plants suggests that they contain some promising active substances such as polyphenols and alkaloids, which could be active as insulin secretagogues and β-cell protectors. In this review, we discuss the potential of several polyphenols, alkaloids and venom peptides from snake, frogs, scorpions and cone snails. These molecules could contribute to the development of new efficient antidiabetic medicines targeting β-cells, which would tackle the progression of the disease.

Novel application of biodegradable chitosan in agriculture: Using green nanopesticides to control Solenopsis invicta

Botanical pesticides are biological pesticides that are environment friendly. However, their instability and short persistence limit their application. In this study, pH sensitive chitosan based rotenone (Rot) nanoparticles (CS/CMCS/Rot-NPs) were prepared using chitosan and carboxymethyl chitosan to take advantage of the acidic nature of the red fire ant midgut. Chitosan based nanoparticles showed photoprotective and slow sustained release effects on Rot and significantly increased the insecticidal activity of Rot against red fire ants. The 24–96hLC50 of CS/CMCS/Rot-NPs against red fire ants was 3.28–6.84 fold that of Rot. The CS/CMCS/Rot-NPs significantly reduced the venom alkaloid content of red fire ants and their living environment and weakened their survival by increasing their survival cost in the ecological environment. Nanotechnology combined with botanical pesticides can be used as a novel, safe, effective, and ecofriendly method to control red fire ants.

Fire ants feed their nestmates with their own venom

Venom secretion is widely used by ants for disease control and more generally as an external surface disinfectant. Here we report evidence that Solenopsis invicta feed their nestmates with their own venom. Venom alkaloids were found in crops and midguts of ants at concentration levels that have previously been reported as effective against various pathogens. These venom alkaloids were found in midguts of the larvae, indicating that trophallaxis must be involved in the transfer of venom, since larvae do not produce alkaloids and they depend on workers to be fed. After the mating flight, the female alates shed their wings, burrow into the soil, and start new colonies. The new queen provided alkaloids to her first batch of larvae in the new colony. Since the crops of female alates contain venom alkaloids donated from their nestmate workers, the transfer of worker alkaloids to new generation occurred. After minim adult workers emerged, they took the role in providing venom to the larvae in the colony. Minim adult workers eventually died out and the normal workers became the venom donors in the colony. Although other functions may be possible, considering the well-known antimicrobial property of venom alkaloids and their detected concentration levels, venom in the digestive system is most likely used as an internal antibiotic by fire ants.

Genetic and Chemical Profiling of Solenopsis spp. (Hymenoptera: Formicidae) Intercepted in Hawaii

Ants intercepted by the Hawaii Department of Agriculture in 2018 and 2019 identified as Solenopsis spp., and possibly S. invicta, were submitted to the United States Department of Agriculture-Agricultural Research Service-Center for Medical, Agricultural and Veterinary Entomology, Imported Fire Ant and Household Insects Research Unit for further taxonomic assessment using genetic and chemical methods. The 2018 sample consisted of 1 worker ant, and the 2019 sample consisted of 3 worker ants. Thoraces were used for genetic assessment and gasters were used for venom analyses. It was determined readily from genetic and chemical analyses that the intercepted ants were not S. invicta or S. richteri. Mitochondrial COX1 gene sequences were compared with previously published data from Solenopsis species. The closest match for the 2018 sample was S. xyloni, which is native to the shipment's origin (San Diego, California, USA). The 2 major alkaloids found were characteristic of the S. geminata species group. In addition, the alkaloid ratio closely matched that of S. xyloni. Therefore, the alkaloid analysis of the 2018 intercepted ant concurs with the genetic data that the ant was an S. xyloni worker. The COX1 gene sequences of the 2019 interceptions most closely matched S. geminata. The corresponding venom alkaloid profile also eliminated S. invicta and S. richteri as possibilities. However, 1 major alkaloid was unusual (Fig. 2) and normally is found only in trace amounts. We suggest the 2019 specimens are an unusual variant of S. geminata.

Characterization of Queen Supergene Pheromone in the Red Imported Fire Ant Using Worker Discrimination Assays

Ants use chemical signals to communicate for various purposes related to colony function. Social organization in the red imported fire ant, Solenopsis invicta, is determined by the Sb supergene, with colonies of the monogyne (single-queen) form lacking the element and colonies of the polygyne (multiple-queen) form possessing it. Polygyne workers accept new reproductive queens in their nest, but only those carrying Sb; young winged queens lacking this genetic element are executed as they mature sexually in their natal nest or as they attempt to enter a foreign nest to initiate reproduction after mating and shedding their wings. It has been suggested that queen supergene genotype status is signaled to workers by unsaturated cuticular hydrocarbons, while queen reproductive status is signaled by piperidines (venom alkaloids). We used high-throughput behavioral assays to study worker acceptance of paper dummies dosed with fractions of extracts of polygyne queens, or blends of synthetic counterparts of queen cuticular compounds. We show that the queen supergene pheromone comprises a blend of monoene and diene unsaturated hydrocarbons. Our assays also reveal that unsaturated hydrocarbons elicit discrimination by polygyne workers only when associated with additional compounds that signal queen fertility. This synergistic effect was obtained with a polar fraction of queen extracts, but not by the piperidine alkaloids, suggesting that the chemical(s) indicating queen reproductive status are compounds more polar than cuticular hydrocarbons but are not the piperidine alkaloids. Our results advance understanding of the role of chemical signaling that is central to the regulation of social organization in an important invasive pest and model ant species.

Fire Ant Venom Alkaloids: Possible Control Measure for Soilborne and Foliar Plant Pathogens.

The purpose of this study was to evaluate fire ant venom alkaloids and an alarm pheromone analog against several plant pathogens, including Botrytis cinerea, Fusarium oxysporum, Phytophthora nicotianae, P. cryptogea, Pseudomonas syringae, Phytopythium citrinum, Rhizoctonia solani, Sclerotonia rolfsii, Xanthomonas axonopodis, and X. campestris. All pathogens were tested against red imported fire ant venom alkaloid extract and alarm pheromone compound for growth inhibition in in vitro assay. The venom alkaloid extract inhibited fungal and oomycete pathogens. Neither of the treatments were effective against bacterial pathogens. Three soilborne pathogens, P. nicotianae, R. solani, F. oxysporum, and one foliar pathogen, B. cinerea were selected for further in-vivo assays on impatiens (Impatiens walleriana ‘Super Elfin XP violet’). Total plant and root weight were higher in venom alkaloid treated plants compared to an inoculated control. The venom alkaloid treatment reduced damping-off, root rot severity, and pathogen recovery in soilborne pathogen inoculated plants. Similarly, venom alkaloid reduced Botrytis blight. However, higher venom rates caused foliar phytotoxicity on plants. Therefore, additional work is needed to evaluate rates of venom alkaloids or formulations to eliminate negative impacts on plants. Overall, these results suggest that red imported fire ant venom alkaloids may provide a basis for new products to control soilborne and foliar plant pathogens.

Unique venom proteins from Solenopsis invicta x Solenopsis richteri hybrid fire ants.

The Solenopsis venom protein 2 transcript was amplified, sequenced, probed, and analyzed from Solenopsis invicta x Solenopsis richteri hybrid ant colonies (hybrids) collected from across Tennessee to determine the extent of introgression of each parent allele (Solenopsis invicta venom protein 2 [Soli2] and Solenopsis richteri venom protein 2 [Solr2]). Chemotaxonomic analyses of venom alkaloids and cuticular hydrocarbons were used to categorize hybrid colonies and their relative relatedness to each parent species. Hybrid colonies were chosen randomly from each chemotaxonomic hybridization category, including “very near S. richteri,” “near S. richteri,” “near S. invicta,” and “very near S. invicta.” Lateral flow immunoassays for detection of the Soli2 and Solr2 venom proteins were largely in agreement with the chemotaxonomic analyses for the very near S. richteri (100% Solr2) and very near S. invicta (80% Soli2, 20% Soli2 + Solr2 detected in the sample) groups, while Soli2 and Solr2 were reported in 60% and 40% in the near S. invicta and near S. richteri chemotaxonomic groups. Analysis of transcripts from the hybrid colonies revealed a sequence with 100% identity to Soli2 (GenBank Accession L09560) and three unique sequences, which we identify as Solenopsis hybrid venom protein 2 (Solh2; GenBank Accession MT150127), Solenopsis hybrid truncated venom protein 2 (Solh2Tr97; Genbank Accession MT150129), and Solenopsis richteri venom protein 2, D to A change at position 69 (Solr2A69; GenBank Accession MT150128). The predicted open reading frame for Solh2 and Solh2Tr97 revealed sequences unique to hybrid ants, with Solh2Tr97an alternatively spliced form. A third unique sequence, Solr2A69, is likely the correct sequence for Solr2, which appears to have been published previously with a sequencing error (GenBank Accession P35776).

Relationship of Imported Fire Ant (Hymenoptera: Formicidae) Integument Coloration to Cuticular Hydrocarbon and Venom Alkaloid Profiles.

Red imported fire ant (Solenopsis invicta Buren; RIFA) and black imported fire ant (Solenopsis richteri Forel (Hymenoptera: Formicidae); BIFA) are considered distinct species with introgression via a reproductively functional hybrid (HIFA). The RIFA and BIFA common names are based on relative coloration. Due to human color perception variation, using color to identify RIFA, BIFA or HIFA is challenging. Fire ant identification traditionally involves molecular or chemical techniques, but a colorimetric test could allow rapid and low-cost identification. In this study, ant integument coloration was measured by spectrophotometer, and color attributes were compared to a combined gas chromatography (GC) index derived from cuticular hydrocarbon and venom alkaloid indices. Significant Pearson Correlation coefficients were found for colony GC index versus color attributes red to green (a*), blue to yellow (b*), chroma (C*), and hue (h*), but not lightness (L*). The RIFA colonies were distinct from BIFA for four of five color attributes and plots of the a*b* and C*h* horizontal axis of the L*a*b* and L*C*h* color spaces. Color attributes for HIFA indices were not distinct from BIFA and RIFA parental species, but HIFA a*b* and C*h* plots were distinct from RIFA and a*b* plots from BIFA. Color attributes a*, b*, and C* increased and h* decreased with GC index in a sinusoidal pattern. In conclusion, most RIFA and BIFA color attributes were distinct and a*b* and C*h* color axes plots had potential to distinguish HIFA from parental species, but coloration of HIFA indices was variable and complicated identifications among HIFA phenotypes, RIFA and BIFA.

Venom Function of a New Species of Megalomyrmex Forel, 1885 (Hymenoptera: Formicidae).

Alkaloids are important metabolites found across a variety of organisms with diverse ecological functions. Of particular interest are alkaloids found in ants, organisms well known for dominating the ecosystems they dwell in. Within ants, alkaloids are found in venom and function as potent weapons against heterospecific species. However, research is often limited to pest species or species with parasitic lifestyles and thus fails to address the broader ecological function of ant venom alkaloids. Here we describe a new species of free-living Megalomyrmex ant: Megalomyrmex peetersi sp. n. In addition, we identify its singular venom alkaloid (trans-2-butyl-5-heptylpyrrolidine) and elucidate the antibiotic and insecticidal functions of its venom. Our results show that Megalomyrmex peetersi sp. n. venom is an effective antibiotic and insecticide. These results are comparable to venom alkaloids found in other ant species, such as Solenopsis invicta. This research provides great insight into venom alkaloid function, and it is the first study to explore these ideas in the Megalomyrmex system.

Venom alkaloids against Chagas disease parasite: search for effective therapies

Chagas disease is an important disease affecting millions of patients in the New World and is caused by a protozoan transmitted by haematophagous kissing bugs. It can be treated with drugs during the early acute phase; however, effective therapy against the chronic form of Chagas disease has yet to be discovered and developed. We herein tested the activity of solenopsin alkaloids extracted from two species of fire ants against the protozoan parasite Trypanosoma cruzi, the aetiologic agent of Chagas disease. Although IC50 determinations showed that solenopsins are more toxic to the parasite than benznidazole, the drug of choice for Chagas disease treatment, the ant alkaloids presented a lower selectivity index. As a result of exposure to the alkaloids, the parasites became swollen and rounded in shape, with hypertrophied contractile vacuoles and intense cytoplasmic vacuolization, possibly resulting in osmotic stress; no accumulation of multiple kinetoplasts and/or nuclei was detected. Overexpressing phosphatidylinositol 3-kinase—an enzyme essential for osmoregulation that is a known target of solenopsins in mammalian cells—did not prevent swelling and vacuolization, nor did it counteract the toxic effects of alkaloids on the parasites. Additional experimental results suggested that solenopsins induced a type of autophagic and programmed cell death in T. cruzi. Solenopsins also reduced the intracellular proliferation of T. cruzi amastigotes in infected macrophages in a concentration-dependent manner and demonstrated activity against Trypanosoma brucei rhodesiense bloodstream forms, which is another important aetiological kinetoplastid parasite. The results suggest the potential of solenopsins as novel natural drugs against neglected parasitic diseases caused by kinetoplastids.

Host preference in parasitic phorid flies: response of Pseudacteon curvatus and P. obtusus to venom alkaloids of native and imported Solenopsis fire ants

Pseudacteon curvatus and P. obtusus are two species of parasitic phorid flies (Diptera: Phoridae), introduced in the southern USA for biological control of Solenopsis fire ants (Hymenoptera: Formicidae). The preference of both parasitoid species (P. curvatus and P. obtusus) for native Solenopsis fire ants (S. geminata complex) versus imported fire ants, S. saevissima complex (S. invicta and S. richteri) was investigated in four-choice olfactometer assays. We hypothesized that differences in the venom alkaloid profiles of fire ant species mediate host preference in phorid flies. In olfactometer bioassays, female P. obtusus showed preference for the venom alkaloid (cis + trans alkaloid mixture) of the imported S. invicta fire ant over the venom alkaloid of the native geminata fire ant. Similarly, P. obtusus showed preference for the venom alkaloid of the S. richteri fire ant over that of the native S. geminata ants. Pseudacteon curvatus also showed preference for the venom alkaloid of the S. invicta and S. richteri fire ants over that of the native geminata ants in separate bioassays. The results suggest that host preference in P. curvatus and P. obtusus is mediated, at least in part, by differences in the venom alkaloid profile of various fire ant species. This finding may explain why Pseudacteon species associated with either the S. geminata or S. saevissima complexes have not switched to the other group despite the overlap of native and imported fire ants in some areas of their distributions.

Pyridine Alkaloids in the Venom of Imported Fire Ants.

Venomous imported fire ants cause significant medical problems. Alkaloids are an important component of imported fire ant venom. Piperidine and piperideine alkaloids have been identified in fire ant venom. In this study, we studied the venom alkaloids of the red imported fire ant, Solenopsis invicta Buren, the black imported fire ant, Solenopsis richteri Forel, and the hybrid, S. invicta × S. richteri. Pyridine alkaloids were detected the first time in fire ants using solid-phase microextraction (SPME) coupled with gas chromatography-mass spectrometry (SPME-GC-MS). The thermal desorption process was manipulated to facilitate the isolation and identification of pyridine alkaloids that were coeluted with piperidine or piperideine alkaloids in GC. After SPME extraction of ant venom, we conducted a series of consecutive GC-MS injections, each with a partial desorption. Hidden pyridine alkaloid peaks were revealed after the overlapping piperidine or piperidiene alkaloid peaks had been desorbed. Using this approach, 10 2-methyl-6-alkyl (or alkenyl)pyridines (1-10) were found the first time in the venom of imported fire ants. Structures of three pyridine alkaloids were confirmed by synthesis, including 2-methyl-6-undecylpyridine (1), 2-methyl-6-tridecylpyridine (7), and 2-methyl-6-pentadecylpyridine (10). We also developed a silica gel column chromatography method to separate the pyridine alkaloids from other alkaloids. Using column chromatography and GC-MS with single ion monitoring at 107 m/z, five pyridine alkaloids were quantified for both workers and female alates of S. invicta and S. richteri.

Fire Ant Venom Alkaloids Inhibit Biofilm Formation

Biofilm formation on exposed surfaces is a serious issue for the food industry and medical health facilities. There are many proposed strategies to delay, reduce, or even eliminate biofilm formation on surfaces. The present study focuses on the applicability of fire ant venom alkaloids (aka ‘solenopsins’, from Solenopsis invicta) tested on polystyrene and stainless steel surfaces relative to the adhesion and biofilm-formation by the bacterium Pseudomonas fluorescens. Conditioning with solenopsins demonstrates significant reduction of bacterial adhesion. Inhibition rates were 62.7% on polystyrene and 59.0% on stainless steel surfaces. In addition, solenopsins drastically reduced cell populations already growing on conditioned surfaces. Contrary to assumptions by previous authors, solenopsins tested negative for amphipathic properties, thus understanding the mechanisms behind the observed effects still relies on further investigation.

Origins and applications of stereoselective sulfenate anion alkylation reactions

Selected sulfenate anions can be alkylated with diastereoselectivities near 9:1 when there is appropriately positioned carbon chirality in the sulfenate structure or in the alkylation electrophile. Both protocols represent a conceptually different approach to β-amino sulfoxides. When vinylic β-amino sulfoxides are cyclized, thiomorpholine S-oxides result, and substrate control over the stereochemistry of alkyl substituents was found. This finding facilitates the synthesis of cis and trans dialkylpyrrolidines, known ant venom alkaloids. A computational model is demonstrated to account for some of the diastereoselection results. The importance of intramolecular interactions of sulfenate constituent atoms with pendent substituents is established.

Worker Size, Geographical Distribution, and Introgressive Hybridization of Invasive Solenopsis invicta and Solenopsis richteri (Hymenoptera: Formicidae) in Tennessee.

Worker size and geographical distribution of red imported fire ants (Solenopsis invicta Buren), black imported fire ants (Solenopsis richteri Forel), and their hybrid (S. invicta × S. richteri) (Hymenoptera: Formicidae) were evaluated from colonies sampled across Tennessee. The fire ant species and hybrid status were determined using cuticular hydrocarbon and venom alkaloid indices obtained from gas chromatography and mass spectrometry. Hybrids were the most common fire ant throughout Tennessee. With the exception of a few isolated S. invicta samples, only hybrids were found in east Tennessee, and hybrids predominated in middle Tennessee. In west Tennessee, mixed populations of S. richteri and hybrids were found. Hybrids were more common in west Tennessee than a survey performed a decade earlier. No statistical differences were detected in the average inter-colonial worker size of S. richteri and hybrids. Likewise, average worker size was not related to geographic location in Tennessee. The similarity in average worker size among hybrid colonies with a wide range of cuticular hydrocarbon and venom alkaloid values suggests introgression was not impacting ant size in colonies sampled throughout Tennessee.

Reaction of Nitrogen‐Radicals with Organometallics Under Ni‐Catalysis: N‐Arylations and Amino‐Functionalization Cascades

AbstractHerein, we report a strategy for the generation of nitrogen‐radicals by ground‐state single electron transfer with organyl–NiI species. Depending on the philicity of the N‐radical, two types of processes have been developed. In the case of nucleophilic aminyl radicals direct N‐arylation with aryl organozinc, organoboron, and organosilicon reagents was achieved. In the case of electrophilic amidyl radicals, cascade processes involving intramolecular cyclization, followed by reaction with both aryl and alkyl organometallics have been developed. The N‐cyclization–alkylation cascade introduces a novel retrosynthetic disconnection for the assembly of substituted lactams and pyrrolidines with its potential demonstrated in the short total synthesis of four venom alkaloids.

Reaction of Nitrogen-Radicals with Organometallics Under Ni-Catalysis: N-Arylations and Amino-Functionalization Cascades.

Herein, we report a strategy for the generation of nitrogen‐radicals by ground‐state single electron transfer with organyl–NiI species. Depending on the philicity of the N‐radical, two types of processes have been developed. In the case of nucleophilic aminyl radicals direct N‐arylation with aryl organozinc, organoboron, and organosilicon reagents was achieved. In the case of electrophilic amidyl radicals, cascade processes involving intramolecular cyclization, followed by reaction with both aryl and alkyl organometallics have been developed. The N‐cyclization–alkylation cascade introduces a novel retrosynthetic disconnection for the assembly of substituted lactams and pyrrolidines with its potential demonstrated in the short total synthesis of four venom alkaloids.

An Alkaloid from Scorpion Venom: Chemical Structure and Synthesis.

While most scorpion venom components identified in the past are peptidic or proteinic in nature, we report here a new alkaloid isolated from the venom of the Mexican scorpion Megacormus gertschi. Nuclear magnetic resonance and mass spectrometric investigations elucidate the structure of the alkaloid as ( Z)- N-(2-(1 H-imidazol-4-yl)ethyl)-3-(4-hydroxy-3-methoxyphenyl)-2-methoxyacrylamide (1). A chemical method of synthesizing this alkaloid is also described. Although abundant in venom, the above alkaloid was not found to have insecticidal activity. Structural analysis suggests that this venom alkaloid might be of potential interest for evaluating its medicinal effect.

Major venom proteins of the fire ant Solenopsis invicta: insights into possible pheromone-binding function from mass spectrometric analysis.

Proteins in the venom of the fire ant Solenopsis invicta have been suggested to function in pheromone binding. Venom from queens and workers contains different isoforms of these proteins, consistent with the differing pheromones they secrete, but questions remain about the venom protein composition and glandular source. We found that the queen venom contains a previously uncharacterized pheromone-binding protein paralogue known as Sol i 2X1. Using imaging mass spectrometry, we located the main venom proteins in the poison sac, implying that pheromones might have to compete with venom alkaloids for binding. Using the known structure of the worker venom protein Sol i 2w, we generated three-dimensional homology models of the worker venom protein Sol i 4.02, and of the two main venom proteins in queens and female alates, Sol i 2q and Sol i 2X1. Surprisingly, the models show that the proteins have relatively small internal hydrophobic binding pockets that are blocked by about 10 amino acids of the C-terminal region. For these proteins to function as carriers of hydrophobic ligands, a conformational change would be required to displace the C-terminal region, somewhat like the mechanism known to occur in the silk moth pheromone-binding protein.