Carrion insects living within the bones of large mammals: insect conservation and forensic entomology implications.

Succession patterns of carrion insects on large mammal's carrion has been widely studied, notably to estimate the post-mortem interval in forensic investigations as accurately as possible. However, little attention has been paid to the carrion insects living inside these bones once a carcass is skeletonized. One very recent study documented flies emerging from pig carcasses, and only scarce authors reported the presence of other carrion insects taking advantage of the bone marrow. We, thus, aimed to (1) estimate the frequency of inner-bone space colonization by carrion insects, with particular attention to bone-skipper flies; (2) identify the insects living inside the carrion bones; and (3) determine whether or not carrion insects found within the bones can successfully exit the bones and complete their development. We extensively sampled 185 large mammals' bones collected from twelve vulture feeding stations and four isolated carcasses in southwest France and northern Spain. Sampled bones were opened, and the insects found inside were identified. For two bones, foramen, i.e., the holes providing a natural entrance and exit to the bone's inner cavity, was monitored with a camera to assess the insect's putative exit. We describe the entomofauna, i.e., the set of insect species, living within the bones, and illustrate insects' ability to exit the bones for their subsequent development and maturity. These results are discussed in the framework of carrion insect conservation and forensic entomology perspectives.

Highly modular pattern in ant-plant interactions involving specialized and non-specialized myrmecophytes.

Because Tachia guianensis (Gentianaceae) is a "non-specialized myrmecophyte" associated with 37 ant species, we aimed to determine if its presence alters the ant guild associated with sympatric "specialized myrmecophytes" (i.e., plants sheltering a few ant species in hollow structures). The study was conducted in a hilly zone of a neotropical rainforest where two specialized myrmecophytes grow at the bottom of the slopes, another at mid-slope, and a fourth on the hilltops. Tachia guianensis, which occurred everywhere, had its own guild of associated ant species. A network analysis showed that its connections with the four other myrmecophytes were rare and weak, the whole resulting in a highly modular pattern of interactions with one module (i.e., subnetwork) per myrmecophyte. Three ant species parasitized three out of the four specialized myrmecophytes (low nestedness noted), but were not or barely associated with T. guianensis that therefore did not influence the parasitism of specialized myrmecophytes.

Hollow Internodes Permit a Neotropical Understory Plant to Shelter Multiple Mutualistic Ant Species, Obtaining Protection and Nutrient Provisioning (Myrmecotrophy).

The Neotropical understory plant Tachia guianensis (Gentianaceae)-known to shelter the colonies of several ant species in its hollow trunks and branches-does not provide them with food rewards (e.g., extrafloral nectar). We tested whether these ants are opportunistic nesters or whether mutualistic relationships exist as for myrmecophytes or plants sheltering ant colonies in specialized hollow structures in exchange for protection from enemies and/or nutrient provisioning (myrmecotrophy). We noted 37 ant species sheltering inside T. guianensis internodes, three of them accounting for 43.5% of the cases. They protect their host plants from leaf-cutting ant defoliation and termite damage because individuals devoid of associated ants suffered significantly more attacks. Using the stable isotope 15N, we experimentally showed that the tested ant species furnish their host plants with nutrients. Therefore, a mutualism exists. However, because it is associated with numerous ant species, T. guianensis can be considered a nonspecialized myrmecophyte.

Ant-lepidopteran associations along African forest edges.

Working along forest edges, we aimed to determine how some caterpillars can co-exist with territorially dominant arboreal ants (TDAAs) in tropical Africa. We recorded caterpillars from 22 lepidopteran species living in the presence of five TDAA species. Among the defoliator and/or nectarivorous caterpillars that live on tree foliage, the Pyralidae and Nymphalidae use their silk to protect themselves from ant attacks. The Notodontidae and lycaenid Polyommatinae and Theclinae live in direct contact with ants; the Theclinae even reward ants with abundant secretions from their Newcomer gland. Lichen feeders (lycaenid; Poritiinae), protected by long bristles, also live among ants. Some lycaenid Miletinae caterpillars feed on ant-attended membracids, including in the shelters where the ants attend them; Lachnocnema caterpillars use their forelegs to obtain trophallaxis from their host ants. Caterpillars from other species live inside weaver ant nests. Those of the genus Euliphyra (Miletinae) feed on ant prey and brood and can obtain trophallaxis, while those from an Eberidae species only prey on host ant eggs. Eublemma albifascia (Erebidae) caterpillars use their thoracic legs to obtain trophallaxis and trophic eggs from ants. Through transfer bioassays of last instars, we noted that herbivorous caterpillars living in contact with ants were always accepted by alien conspecific ants; this is likely due to an intrinsic appeasing odor. Yet, caterpillars living in ant shelters or ant nests probably acquire cues from their host colonies because they were considered aliens and killed. We conclude that co-evolution with ants occurred similarly in the Heterocera and Rhopalocera.

How territoriality and host-tree taxa determine the structure of ant mosaics.

Very large colonies of territorially dominant arboreal ants (TDAAs), whose territories are distributed in a mosaic pattern in the canopies of many tropical rainforests and tree crop plantations, have a generally positive impact on their host trees. We studied the canopy of an old Gabonese rainforest (ca 4.25 ha sampled, corresponding to 206 "large" trees) at a stage just preceding forest maturity (the Caesalpinioideae dominated; the Burseraceae were abundant). The tree crowns sheltered colonies from 13 TDAAs plus a co-dominant species out of the 25 ant species recorded. By mapping the TDAAs' territories and using a null model co-occurrence analysis, we confirmed the existence of an ant mosaic. Thanks to a large sampling set and the use of the self-organizing map algorithm (SOM), we show that the distribution of the trees influences the structure of the ant mosaic, suggesting that each tree taxon attracts certain TDAA species rather than others. The SOM also improved our knowledge of the TDAAs' ecological niches, showing that these ant species are ecologically distinct from each other based on their relationships with their supporting trees. Therefore, TDAAs should not systematically be placed in the same functional group even when they belong to the same genus. We conclude by reiterating that, in addition to the role played by TDAAs' territorial competition, host trees contribute to structuring ant mosaics through multiple factors, including host-plant selection by TDAAs, the age of the trees, the presence of extrafloral nectaries, and the taxa of the associated hemipterans.

Urbanization and Water Containers Influence the Mosquito Community with Consequences for Aedes aegypti.

We aimed to determine how the degree of urbanization in a Neotropical city influences Aedes aegypti (L.), a pantropical vector of urban yellow fever, dengue, Zika and Chikungunia, via other mosquito species, whether they are competitors or predators, native to the area or invasive. We conducted experiments twice a month during one year in the city of Kourou, French Guiana, on three sites characterized by increasing percentages of imperviousness (i.e., 0.65%, 33.80% and 86.60%). These sites were located in a ≈5 ha forest fragment, a residential area with gardens, and in the older part of the city, respectively, and correspond to slightly, moderately and highly urbanized sites. There, we monitored twice a month during one year a total of 108 mosquito communities inhabiting four types of containers (i.e., a tank bromeliad, dry stumps of bamboo, ovitraps and car tires) installed in a random block design. In the tanks of the bromeliad, likely due to the acidity of the water, the immatures of native mosquito species prevailed, particularly Wyeomyia pertinans (Williston) in the slightly urbanized site. The general pattern was very similar in the three other types of containers where Limatus durhamii Théobald dominated in the slightly urbanized site, so that the abundance of Ae. aegypti immatures was low compared to those of native species. Yet, Ae. aegypti strongly dominated in the two more urbanized sites. These findings open up perspectives for vector management, including the conservation and/or the augmentation of natural enemies through modifications to landscape features.

Foraging by predatory ants: A review.

In this review, we show that predatory ants have a wide range of foraging behavior, something expected given their phylogenetic distance and the great variation in their colony size, life histories, and nesting habitats as well as prey diversity. Most ants are central-place foragers that detect prey using vision and olfaction. Ground-dwelling species can forage solitarily, the ancestral form, but generally recruit nestmates to retrieve large prey or a group of prey. Typically, ants are omnivorous, but some species are strict predators preying on detritivorous invertebrates or arthropod eggs, while those specialized on termites or other ants often have scouts that localize their target and then trigger a raid. They can use compounds that ease this task, including chemical insignificance, mimicry, and venoms triggering submissive behavior. Army ants include 8 Dorylinae and some species from other subfamilies, all having wingless queens and forming raids. Dorylinae from the Old World migrate irregularly to new nesting sites. The foraging of most New World species that prey on the brood of other ants is regulated by their biological cycle that alternates between a "nomadic phase" when the colony relocates between different places and a "stationary phase" when the colony stays in a bivouac constituting a central place. Among arboreal ants, dominant species forage in groups, detecting prey visually, but can use vibrations, particularly when associated with myrmecophytes. Some species of the genera Allomerus and Azteca use fungi to build a gallery-shaped trap with small holes under which they hide to ambush prey.

Altruism during predation in an assassin bug.

Zelus annulosus is an assassin bug species mostly noted on Hirtella physophora, a myrmecophyte specifically associated with the ant Allomerus decemarticulatus known to build traps on host tree twigs to ambush insect preys. The Z. annulosus females lay egg clutches protected by a sticky substance. To avoid being trapped, the first three instars of nymphs remain grouped in a clutch beneath the leaves on which they hatched, yet from time to time, they climb onto the upper side to group ambush preys. Long-distance prey detection permits these bugs to capture flying or jumping insects that alight on their leaves. Like some other Zelus species, the sticky substance of the sundew setae on their forelegs aids in prey capture. Group ambushing permits early instars to capture insects that they then share or not depending on prey size and the hunger of the successful nymphs. Fourth and fifth instars, with greater needs, rather ambush solitarily on different host tree leaves, but attract siblings to share large preys. Communal feeding permits faster prey consumption, enabling small nymphs to return sooner to the shelter of their leaves. By improving the regularity of feeding for each nymph, it likely regulates nymphal development, synchronizing molting and subsequently limiting cannibalism.

Mutualistic interactions between ants and fungi: A review.

The large amount of dead plant biomass caused by the final extinction events triggered a fungi proliferation that mostly differentiated into saprophytes degrading organic matter; others became parasites, predators, likely commensals, and mutualists. Among the last, many have relationships with ants, the most emblematic seen in the Neotropical myrmicine Attina that cultivate Basidiomycota for food. Among them, leaf-cutting, fungus-growing species illustrate an ecological innovation because they grow fungal gardens from fresh plant material rather than arthropod frass and plant debris. Myrmecophytes shelter "plant-ants" in hollow structures, the domatia, whose inner walls are lined with thin-walled Ascomycota hyphae that, in certain cases, are eaten by the ants, showing a form of convergence. Typically, these Ascomycota have antibacterial properties illustrating cases of farming for protection. Ant gardens, or mutualistic associations between certain ant species and epiphytes, shelter endophytic fungi that promote the growth of the epiphytes. Because the cell walls of certain Ascomycota hyphae remain sturdy after the death of the mycelium, they form resistant fibers used by ants to reinforce their constructions (e.g., galleries, shelters for tended hemipterans, and carton nests). Thus, we saw cases of "true" fungal agriculture involving planting, cultivating, and harvesting Basidiomycota for food with Attina. A convergence with "plant-ants" feeding on Ascomycota whose antibacterial activity is generally exploited (i.e., farming for protection). The growth of epiphytes was promoted by endophytic fungi in ant gardens. Finally, farming for structural materials occurred with, in one case, a leaf-cutting, fungus-growing ant using Ascomycota fibers to reinforce its nests.

Long-term phytoplankton dynamics in the Zeeschelde estuary (Belgium) are driven by the interactive effects of de-eutrophication, altered hydrodynamics and extreme weather events.

We studied how changing human impacts affected phytoplankton dynamics in the freshwater and brackish tidal reaches of the Zeeschelde estuary (Belgium) between 2002 and 2018. Until the early 2000s, the Zeeschelde was heavily polluted due to high wastewater discharges. By 2008, water quality had improved, resulting in lower nutrient concentrations and higher oxygen levels. Since 2009, however, increased dredging activities resulted in altered hydrodynamics and increased suspended sediment concentration. The combined effects of these environmental changes were reflected in three marked transitions in phytoplankton community composition. Assemblages were dominated by Thalassiosirales and green algae (especially Scenedesmaceae) until 2003. The period 2003-2011 was characterized by the wax and wane of the centric diatoms Actinocyclus and Aulacoseira, while in the period 2012-2018 Thalassiosirales and Cyanobacteria became dominant, the latter mainly imported from the tributaries. Phytoplankton biomass increased sharply in 2003, after which there was a gradual decline until 2018. By 2018, the timing of the growing season had advanced with about one month compared to the start of the study, probably as a consequence of climate warming and intensified zooplankton grazing pressure. Our study shows that de-eutrophication (during the 2000s) and morphological interventions in the estuary (in the 2010s) were dominant drivers of phytoplankton dynamics but that the main shifts in community composition were triggered by extreme weather events, suggesting significant resistance of autochthonous communities to gradual changes in the environment.

When attempts at robbing prey turn fatal.

Because group-hunting arboreal ants spread-eagle insect prey for a long time before retrieving them, these prey can be coveted by predatory flying insects. Yet, attempting to rob these prey is risky if the ant species is also an effective predator. Here, we show that trying to rob prey from Azteca andreae workers is a fatal error as 268 out of 276 potential cleptobionts (97.1 %) were captured in turn. The ant workers hunt in a group and use the "Velcro®" principle to cling firmly to the leaves of their host tree, permitting them to capture very large prey. Exceptions were one social wasp, plus some Trigona spp. workers and flies that landed directly on the prey and were able to take off immediately when attacked. We conclude that in this situation, previously captured prey attract potential cleptobionts that are captured in turn in most of the cases.

Breaking the silence: how shedding light on the bone-skipper fly Thyreophora cynophila (Diptera: Piophilidae) demonstrated it still has a large distribution area in the Pyrenees mountains, France.

BACKGROUND: Thyreophora cynophila (Panzer, 1798) is an iconic species of the European entomofauna. This winter-flying necrophagous fly was considered long extinct in Europe, before being discovered in Spain in 2010 and re-discovered in France in 2020, with a unique locality in Saint-Paul-de-Jarrat (Ariège, southern France). NEW INFORMATION: After bringing this species to the attention of people that are active in nature during the winter, including hunters, skilled naturalists, nature lovers and professional naturalists, we gathered seven new occurrence data for this species at six locations on the French flanks of the Pyrenees mountains. Those data considerably extend the known distribution of the species in Europe and allows mapping the first approximate extent of occurrence for this species in France.

An uneasy alliance: a nesting association between aggressive ants and equally fierce social wasps.

Although the Neotropical territorially dominant arboreal ant Azteca chartifex Forel is very aggressive towards any intruder, its populous colonies tolerate the close presence of the fierce polistine wasp Polybia rejecta (F.). In French Guiana, 83.33% of the 48 P. rejecta nests recorded were found side by side with those of A. chartifex. This nesting association results in mutual protection from predators (i.e., the wasps protected from army ants; the ants protected from birds). We conducted field studies, laboratory-based behavioral experiments and chemical analyses to elucidate the mechanisms allowing the persistence of this association. Due to differences in the cuticular profiles of the two species, we eliminated the possibility of chemical mimicry. Also, analyses of the carton nests did not reveal traces of marking on the envelopes. Because ant forager flows were not perturbed by extracts from the wasps' Dufour's and venom glands, we rejected any hypothetical action of repulsive chemicals. Nevertheless, we noted that the wasps "scraped" the surface of the upper part of their nest envelope using their mandibles, likely removing the ants' scent trails, and an experiment showed that ant foragers were perturbed by the removal of their scent trails. This leads us to use the term "erasure hypothesis." Thus, this nesting association persists thanks to a relative tolerance by the ants towards wasp presence and the behavior of the wasps that allows them to "contain" their associated ants through the elimination of their scent trails, direct attacks, "wing-buzzing" behavior and ejecting the ants.

An arboreal spider protects its offspring by diving into the water of tank bromeliads.

Cupiennius salei (Ctenidae) individuals frequently live in association with tank bromeliads, including Aechmea bracteata, in Quintana Roo (Mexico). Whereas C. salei females without egg sacs hunt over their entire host plant, females carrying egg sacs settle above the A. bracteata reservoirs they have partially sealed with silk. There they avoid predators that use sight to detect their prey, as is known for many bird species. Furthermore, if a danger is more acute, these females dive with their egg sacs into the bromeliad reservoir. An experiment showed that this is not the case for males or females without egg sacs. In addition to the likely abundance of prey found therein, the potential of diving into the tank to protect offspring may explain the close association of this spider with bromeliads. These results show that, although arboreal, C. salei evolved a protective behavior using the water of tank bromeliads to protect offspring.

Ants impact the composition of the aquatic macroinvertebrate communities of a myrmecophytic tank bromeliad.

In an inundated Mexican forest, 89 out of 92 myrmecophytic tank bromeliads (Aechmea bracteata) housed an associated ant colony: 13 sheltered Azteca serica, 43 Dolichoderus bispinosus, and 33 Neoponera villosa. Ant presence has a positive impact on the diversity of the aquatic macroinvertebrate communities (n=30 bromeliads studied). A Principal Component Analysis (PCA) showed that the presence and the species of ant are not correlated to bromeliad size, quantity of water, number of wells, filtered organic matter or incident radiation. The PCA and a generalized linear model showed that the presence of Azteca serica differed from the presence of the other two ant species or no ants in its effects on the aquatic invertebrate community (more predators). Therefore, both ant presence and species of ant affect the composition of the aquatic macroinvertebrate communities in the tanks of A. bracteata, likely due to ant deposition of feces and other waste in these tanks.

Aquatic life in Neotropical rainforest canopies: Techniques using artificial phytotelmata to study the invertebrate communities inhabiting therein.

In Neotropical rainforest canopies, phytotelmata ("plant-held waters") shelter diverse aquatic macroinvertebrate communities, including vectors of animal diseases. Studying these communities is difficult because phytotelmata are widely dispersed, hard to find from the ground and often inaccessible. We propose here a method for placing in tree crowns "artificial phytotelmata" whose size and shape can be tailored to different research targets. The efficacy of this method was shown while comparing the patterns of community diversity of three forest formations. We noted a difference between a riparian forest and a rainforest, whereas trees alongside a dirt road cutting through that rainforest corresponded to a subset of the latter. Because rarefied species richness was significantly lower when the phytotelmata were left for three weeks rather than for six or nine weeks, we recommend leaving the phytotelmata for twelve weeks to permit predators and phoretic species to fully establish themselves.

The predatory behavior of the Neotropical social wasp Polybia rejecta.

We experimentally studied the predatory behavior of Polybia rejecta (Vespidae, Polistinae, Epiponini) towards 2-88 mm-long insects attracted to a UV light trap. Foragers, which began to hunt at 6:30, selected 4-14 mm-long prey insects. Prey detection by sight by hovering wasps was confirmed using decoys. After the wasps landed and walked along a sinuous path, prey were detected by contact or from a distance (1-3cm). This was followed by seizure, stinging (contrarily to most other known cases), prey manipulation and retrieval. Prey that flew off might be caught in flight. The prey load, representing 30.7% of a forager's weight, was optimized by capturing up to six small prey or two medium-sized prey successively (both of which might be consumed in situ). The foragers cut off the wings of larger prey or cut them into two pieces and returned to gather the second piece. The handling time increased exponentially with the weight of the prey. Partial loading (i.e., retrieving a load much inferior to the maximum possible) was likely related to social facilitation, a form of nest-based recruitment that was demonstrated through the experimental elimination of local enhancement by removing foragers (both mechanisms favor the exploitation of favorable patches).

A cuckoo-like parasitic moth leads African weaver ant colonies to their ruin.

In myrmecophilous Lepidoptera, mostly lycaenids and riodinids, caterpillars trick ants into transporting them to the ant nest where they feed on the brood or, in the more derived "cuckoo strategy", trigger regurgitations (trophallaxis) from the ants and obtain trophic eggs. We show for the first time that the caterpillars of a moth (Eublemma albifascia; Noctuidae; Acontiinae) also use this strategy to obtain regurgitations and trophic eggs from ants (Oecophylla longinoda). Females short-circuit the adoption process by laying eggs directly on the ant nests, and workers carry just-hatched caterpillars inside. Parasitized colonies sheltered 44 to 359 caterpillars, each receiving more trophallaxis and trophic eggs than control queens. The thus-starved queens lose weight, stop laying eggs (which transport the pheromones that induce infertility in the workers) and die. Consequently, the workers lay male-destined eggs before and after the queen's death, allowing the colony to invest its remaining resources in male production before it vanishes.