Morphology of the mandibular gland of the ant Paraponera clavata

Billy Bennett

The ant Paraponera clavata (Fabricius, 1775) is the only extant species of Paraponerinae and is widely distributed in Brazilian forests. Aspects of its biology are documented extensively in the literature; however, knowledge of P. clavata internal morphology, specifically of exocrine glands, is restricted to the venom apparatus. The objective of this study was to describe the mandibular gland morphology of P. clavata workers. The mandibular gland is composed of a reservoir connected to a cluster of Type III secretory cells with cytoplasm rich in mitochondria and lipid droplets, similar to that of other ants. Notably, the glandular secretion is rich in protein and has a solid aspect. This is the first morphological description of the mandibular gland of P. clavata. RESEARCH HIGHLIGHTS: This study presents the morphological description of the mandibular gland of Paraponera clavata (Hymenoptera: Paraponerinae). Singular characteristics of the gland are described: the glandular secretion is rich in protein and has a solid aspect.

Combined Peptidomic and Proteomic Analysis of Electrically Stimulated and Manually Dissected Venom from the South American Bullet Ant Paraponera clavata.

Ants have evolved venoms rich in peptides and proteins used for predation, defense, and communication. However, they remain extremely understudied due to the minimal amount of venom secreted by each ant. The present study investigated the differences in the proteome and peptidome of the venom from the bullet ant, Paraponera clavata. Venom samples were collected from a single colony either by manual venom gland dissection or by electrical stimulation and were compared using proteomic methods.

Venom proteins were separated by 2D-PAGE and identified by nanoLC-ESI-QTOF MS/MS. Venom peptides were initially separated using C18 reversed-phase high-performance liquid chromatography, then analyzed by MALDI-TOF MS. The proteomic analysis revealed numerous proteins that could be assigned a biological function (total 94), mainly as toxins, or roles in cell regulation and transport. This investigation found that ca. 73% of the proteins were common to venoms collected by the two methods. The peptidomic analysis revealed a large number of peptides (total 309) but with <20% shared by the two collection methods. There was also a marked difference between venoms obtained by venom gland dissection from different ant colonies. These findings demonstrate the rich composition and variability of P. clavata venom.

Foraging energetics of the ant, Paraponera clavata.

The energy currencies used by foraging animals are expected to relate to the energy costs and benefits of resource collection. However, actual costs of foraging are rarely measured. We measured the ratio of energetic benefit relative to cost (B/C) during foraging for the giant tropical ant, Paraponera clavata. The B/C ratio was 3.9 for nectar-foragers and 67 for insect prey foragers. In contrast, the B/C ratio during foraging for seed harvester ants (Pogonomyrmex occidentalis) is over 1000, demonstrating that the B/C ratio can vary widely among ants.

The B/C ratio was 300 times lower for nectar-foraging Paraponera than for the seed-harvesting Pogonomyrmex because of: (1) a 5-fold lower energetic benefit per trip, (2) a 10-fold greater cost due to longer foraging distances, and (3) a 6-fold greater energy cost per meter due to larger body size. For Paraponera daily colonial energy intake rates are similar to expeditures and may limit colony growth and reproduction. In contrast, for Pogonomyrmex energy intake rates are an order of magnitude higher than estimated costs, suggesting that Pogonomyrmex colonies are unlikely to be limited by short-term energy intake. We suggest that variation in individual B/C ratios may explain why the foraging behavior of Paraponera but not Pogonomyrmex appears sensitive to foraging costs.

Distance effects on resource profitability and recruitment in the giant tropical ant, Paraponera clavata

We examine how cost and benefit components of resource profitability affect recruitment in the giant tropical ant, Paraponera clavata. To vary resource profitability, we changed the quantity of artificial nectar baits presented to foragers and the distance of nectar baits from the nest. Both distance to and amount of resource affected quantitative aspects of recruitment. At increased distances foragers were less likely to recruit, and fewer workers were recruited to the resource area. The amount of nectar affected the tendency of foragers to recruit, but had no effect on the number of ants recruited. Variation in resource distance was also associated with qualitative changes in recruitment strategy. Foragers at distant sites recruited from the canopy rather than from the nest, and often transferred nectar to other workers for transport to the nest.

Nectar transfer and extra-nidal recruitment significantly reduced the time required for resource collection. It may also have increased the ability of workers to specialize in specific foraging tasks. A portion of the colony’s foraging force specialized spatially by remaining in distant foraging areas without returning to the nest. The flexible recruitment system of P. clavata increases colonial net energetic gain rates by concentrating foraging effort on resources yielding the highest net energetic rewards, and increases the competitive abilities of individual colonies at resource sites by decreasing collection times.

A reexamination of poneratoxin from the venom of the bullet ant Paraponera clavata.

In 1991, Piek et al. [45] described a voltage-gated sodium channel (VGSC) modifier from “bullet ant” (Paraponera clavata) venom they called poneratoxin (PoTx). Using UV chromatography and Edman degradation they showed two “identical peptides” of 25 residues. We reinvestigated PoTx using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-TMS). De novo sequencing showed the two peptides were actually structurally different peptides: the originally described PoTx and a glycyl pro-peptide (glycyl-PoTx) that lacks C-terminus amidation.

We examined P. clavata venom from different geographical locations and discovered two additional PoTx analogs: an A23E substitution analog and a D22N; A23V substitutions analog. We tested PoTx and these three natural analogs on the mammalian sensory voltage-gated sodium channel, Nav1.7, using whole-cell voltage-clamp. PoTx and each analog induced slowly activating currents in response to small depolarizing steps and sustained currents due to blockade of channel inactivation, similar to that described previously in skeletal muscle [19]. Glycyl-PoTx had the same potency and efficacy as PoTx. A23E PoTx, with a decrease in both C-terminal net positive charge and hydrophobicity, had an eight-fold reduction in potency compared to PoTx. In contrast, the D22N; A23V PoTx, with an increase in both C-terminal net positive charge and hydrophobicity, had a nearly five-fold increase in potency compared to PoTx. We found that changes in PoTx C-terminus caused a significant change in PoTx potency.

Distribution and dietary regulation of an associated facultative Rhizobiales-related bacterium in the omnivorous giant tropical ant, Paraponera clavata

We document a facultative Bartonella-like Rhizobiales bacterium in the giant tropical ant, Paraponera clavata. In a lowland tropical rainforest in Costa Rica, 59 colonies were assayed for the prevalence of the Bartonella-like bacterium (BLB), 14 of which were positive. We addressed three questions: First, how does the prevalence of BLB within colonies vary with environmental conditions? Second, how does diet affect the prevalence of BLB in P. clavata? Third, how does the distribution of BLB among colonies reflect ambient differences in food resources and foraging habits? A variety of environmental variables that may be predictive of the presence of BLB were measured, and diet manipulations were conducted to test whether the prevalence of BLB responded to supplemental carbohydrate or prey.

Paraponera clavata Poneratoxin
1-CSB-BP340426PCP	Cusabio	1008.00 EUR 398.40 EUR 2604.00 EUR 1362.00 EUR 1894.80 EUR 564.00 EUR	100ug 10ug 1MG 200ug 500ug 50ug
Recombinant Aspergillus Clavatus AK1 Protein (aa 1-257) [His]
VAng-Wyb3747-1mg	Creative Biolabs	1 mg	5078.4 EUR
Recombinant Aspergillus Clavatus H2A Protein (aa 2-133) [His]
VAng-Wyb3775-1mg	Creative Biolabs	1 mg	4488 EUR
Recombinant Aspergillus Clavatus H2A.Z Protein (aa 1-138) [His]
VAng-Wyb3776-1mg	Creative Biolabs	1 mg	4514.4 EUR
Recombinant Aspergillus Clavatus CHZ1 Protein (aa 1-115) [His]
VAng-Wyb3755-1mg	Creative Biolabs	1 mg	4407.6 EUR
Recombinant Aspergillus Clavatus CSM3 Protein (aa 1-267) [His]
VAng-Wyb3757-1mg	Creative Biolabs	1 mg	5131.2 EUR
Recombinant Aspergillus Clavatus CTU2 Protein (aa 1-374) [His]
VAng-Wyb3758-1mg	Creative Biolabs	1 mg	5636.4 EUR
Recombinant Aspergillus Clavatus DDI1 Protein (aa 1-404) [His]
VAng-Wyb3759-1mg	Creative Biolabs	1 mg	5778 EUR
Recombinant Aspergillus Clavatus DML1 Protein (aa 1-499) [His]
VAng-Wyb3760-1mg	Creative Biolabs	1 mg	6226.8 EUR
Recombinant Aspergillus Clavatus DRE2 Protein (aa 1-310) [His]
VAng-Wyb3761-1mg	Creative Biolabs	1 mg	5336.4 EUR
Recombinant Aspergillus Clavatus EFG1 Protein (aa 1-315) [His]
VAng-Wyb3762-1mg	Creative Biolabs	1 mg	5359.2 EUR
Recombinant Aspergillus Clavatus Egd2 Protein (aa 1-204) [His]
VAng-Wyb3763-1mg	Creative Biolabs	1 mg	4827.6 EUR
Recombinant Aspergillus Clavatus MED7 Protein (aa 1-260) [His]
VAng-Wyb3789-1mg	Creative Biolabs	1 mg	5095.2 EUR
Recombinant Aspergillus Clavatus MED8 Protein (aa 1-260) [His]
VAng-Wyb3790-1mg	Creative Biolabs	1 mg	5067.6 EUR
Recombinant Aspergillus Clavatus MMM1 Protein (aa 44-494) [His]
VAng-Wyb3792-1mg	Creative Biolabs	1 mg	5998.8 EUR
Recombinant Aspergillus Clavatus ACLA_005680 Protein (aa 1-471) [His]
VAng-Wyb3745-1mg	Creative Biolabs	1 mg	6098.4 EUR
Recombinant Aspergillus Clavatus ACLA_026210 Protein (aa 1-71) [His]
VAng-Wyb3746-1mg	Creative Biolabs	1 mg	4197.6 EUR
Recombinant Aspergillus Clavatus ATG3 Protein (aa 1-358) [His]
VAng-Wyb3749-1mg	Creative Biolabs	1 mg	5563.2 EUR

The ambient frequency of BLB is much higher in young secondary forests, but is nearly absent from older secondary forests. The prevalence of BLB inside field colonies increased over the duration of a 2-week carbohydrate supplementation; however, water and prey supplementation did not alter the prevalence of BLB. The diets of the colonies located in young secondary forest, compared to other habitats, have a diet richer in carbohydrates and lower in prey. The abundance of carbohydrate, or the relative lack of N, in a colony’s diet influences the occurrence of the BLB microbe in P. clavata. As experimental diet manipulations can affect the facultative presence of an N-cycling microbe, a consistent diet shift in diet may facilitate the emergence of tighter symbioses.