Fecundity and longevity of Argentine ant (Hymenoptera: Formicidae) queens in response to irradiation

Irradiation is a post‐harvest quarantine treatment option to control ants and other hitchhiker pests on fresh horticultural products traded between countries. As little is known about irradiation effects on ants, radiotolerance of the Argentine ant, Linepithema humile (Mayr) (Hymenoptera: Formicidae: Dolichoderinae), was studied to determine a dose sufficient for its control. Queens collected from Buenos Aires, Argentina, were irradiated with 30, 60, 90 Gy or left untreated as controls, and then followed for 8 weeks to evaluate their survival and fecundity. Overall queen survival and brood viability decreased with increasing irradiation dose. The number of eggs was reduced by 50%, 69% and 56% in the 30, 60 and 90 Gy doses, respectively, compared with untreated control queens. The percentage of eggs that developed into larvae decreased from 41.1% in the control to 22.5%, 1.4%, and 0% in the 30, 60, and 90 Gy treatments, respectively. Thus, the number of larvae was reduced by 69% in the 30 Gy treatment compared with the control, only one larva was observed in the 60 Gy treatment, and none in the 90 Gy treatment. Only one pupa was observed in the 30 Gy treatment and none in the 60 and 90 Gy treatments during the 8‐week experiment. Queens irradiated with 60 and 90 Gy had significantly reduced longevity compared with queens treated with lower doses or untreated queens. Radiation dose ≥90 Gy stopped brood development in Argentine ant queens and should be sufficient as a phytosanitary treatment. The radiotolerance of Argentine ant appears to be similar to that of two other important invasive ants.     

Let them eat termites—prey‐baiting provides effective control of Argentine ants,Linepithema humile,in a biodiversity hotspot

Invasive ants threaten biodiversity, ecosystem services and agricultural systems. This study evaluated a prey‐baiting approach for managing Argentine ants in natural habitat invaded by Argentine ants. Blackmound termites (Amitermes hastatus) were topically exposed to fipronil and presented to Argentine ants (Linepithema humile). In laboratory assays, L. humile colonies were offered fipronil‐treated termites within experimental arenas. The termites were readily consumed, and results demonstrate that a single termite topically treated with 590 ng fipronil is capable of killing at least 500 L. humile workers in 4 days. Field studies were conducted in natural areas invaded by L. humile. Fipronil‐treated termites scattered within experimental plots provided rapid control of L. humile and ant densities throughout the treated plots declined by 98 ± 5% within 21 days. Results demonstrate that the prey‐baiting approach is highly effective against L. humile and may offer an effective alternative to traditional bait treatments. Furthermore, prey‐baiting offers environmental benefits by delivering substantially less toxicant to the environment relative to current control methods which rely on commercial bait formulations and may offer greater target specificity.     

A single lycaenid caterpillar gets an ant‐constructed shelter and uninterrupted ant attendance

Ant‐lycaenid associations range from mutualism to parasitism and the caterpillars of some species of lycaenids are reported to enter ant nests for shelter, diapause, or pupation. The present study aimed to examine the nature of the association between Euchrysops cnejus (Fabricius) (Lepidoptera: Lycaenidae) and Camponotus compressus (Fabricius) (Hymenoptera: Formicidae) worker ants on the extrafloral nectary‐bearing cowpea plant, Vigna unguiculata (L.) Walp. (Fabaceae). The abundance patterns of the ants and the lycaenid caterpillars together with the spatial patrolling patterns of the ants on the plants revealed that ant abundance increased with the occurrence of the lycaenid caterpillars and the ants preferred the lycaenids over the extrafloral nectar. Camponotus compressus worker ants constructed a shelter at the cowpea plant base after interaction with one or more lycaenid caterpillar(s) and tended the caterpillars and pupae till the emergence of the butterfly. The ant‐constructed shelters (ACSs) inhabited by the minor caste workers (13 ± 1.3 ants per ACS), were utilized by the caterpillars to undergo pupation. The ants confined their activities predominantly to tending the pod‐feeding caterpillars and the solitary pupa within each ACS. It appears that the behavior of the tending worker ants is modulated by the lycaenid vulnerable stages.     

Plant‐derived differences in the composition of aphid honeydew and their effects on colonies of aphid‐tending ants

In plant–ant–hemipteran interactions, ants visit plants to consume the honeydew produced by phloem‐feeding hemipterans. If genetically based differences in plant phloem chemistry change the chemical composition of hemipteran honeydew, then the plant's genetic constitution could have indirect effects on ants via the hemipterans. If such effects change ant behavior, they could feed back to affect the plant itself. We compared the chemical composition of honeydews produced by Aphis nerii aphid clones on two milkweed congeners, Asclepias curassavica and Asclepias incarnata, and we measured the responses of experimental Linepithema humile ant colonies to these honeydews. The compositions of secondary metabolites, sugars, and amino acids differed significantly in the honeydews from the two plant species. Ant colonies feeding on honeydew derived from A. incarnata recruited in higher numbers to artificial diet, maintained higher queen and worker dry weight, and sustained marginally more workers than ants feeding on honeydew derived from A. curassavica. Ants feeding on honeydew from A. incarnata were also more exploratory in behavioral assays than ants feeding from A. curassavica. Despite performing better when feeding on the A. incarnata honeydew, ant workers marginally preferred honeydew from A. curassavica to honeydew from A. incarnata when given a choice. Our results demonstrate that plant congeners can exert strong indirect effects on ant colonies by means of plant‐species‐specific differences in aphid honeydew chemistry. Moreover, these effects changed ant behavior and thus could feed back to affect plant performance in the field.     

Demography and sustainable management of two fiber‐producing Astrocaryum palms in Colombia

The spear leaves of the palms Astrocaryum chambira and A. standleyanum have been traditionally used by Colombian indigenous communities as a source of fiber for handicraft production. Traditional management practices, including destructive harvest, have reduced population sizes of both species. We monitored a population of A. chambira in the Amazon, and one of A. standleyanum at the Pacific lowlands of Colombia. We then constructed integral projection models (IPM) to evaluate the transient population dynamics of populations under different exploitation regimes. Our results show that during the next 50 years the population of A. standleyanum will grow at an annual rate of 2.0 percent, and that of A. chambira at a rate of 0.8 percent. However, projected population growth is highly sensitive to harvest in both species: a destructive harvest of 5 percent of all usable individuals (subadults and adults) would cease population growth, while a 10 percent harvest intensity would cause populations to decrease by 0.5–0.6 percent annually. Our simulations further indicate that management practices associated with indigenous slash‐and‐burn agriculture would reduce fiber production, whereas caring for seedlings would increase population growth and fiber production in the coming decades. In order to sustain viable populations of both species and maintain a steady fiber supply, it is vital to prevent destructive harvest practices, and to leave some forest areas untouched, where populations can regenerate and act as a source of seedlings for intervened areas.     

Plant‐based compounds with potential as push‐pull stimuli to manage behavior of leaf‐cutting ants

Leaf‐cutting ants are a serious pest of young forestry plantations. Currently, the main control method is the use of broad‐spectrum insecticides, which have a negative effect on non‐target organisms and the environment. In this work, plant‐based compounds were evaluated in laboratory assays with Acromyrmex ambiguus Emery (Hymenoptera: Formicidae) for their potential use as repellent and attractant stimuli to be used in a push‐pull strategy. Farnesol, a sesquiterpene present in many essential oils, was tested as a repellent at doses of 10, 50, and 100 mg. Its distance of action was studied by comparing the repellent effect of farnesol in a situation in which ants had to touch the farnesol in order to reach the food source in comparison to when ants could reach the food source without getting into direct contact with it. Different parts of the orange fruit (pulp and peel) were evaluated and compared as attractants, given that citrus‐based baits are among the most popular attractants used. Results from laboratory bioassays indicated that farnesol is repellent at doses of 50 mg and acts upon contact or at a very short distance. Furthermore, orange pulp was more attractive than the peel, and volatile compounds were highly responsible for the attraction. When both stimuli were tested simultaneously in a laboratory experiment, repellency of farnesol was enhanced in the presence of orange pulp odor. When tested in a field push‐pull experiment, the results also showed a good repellent effect of farnesol as well as an attractant effect of the orange pulp. These results encourage long‐term studies with these substances in a field setting and suggest that repellents can be enhanced by the use of attractants to manage leaf‐cutting ants behavior.     

Effects of land‐use changes on ecosystem services: decrease in ant predation in human‐dominated landscapes in central Brazil

Anthropogenic disturbances often affect the abundance and diversity of ants (Hymenoptera: Formicidae) but relatively few studies have explored the implications of such changes on the ecosystem services mediated by these insects. Here, we evaluated how the transformation of Cerrado savanna habitats into crop plantations affects the abundance, diversity, and the predatory activity of ants. A survey of the ant faunas foraging above‐ and belowground was performed in six crop and six non‐crop (i.e., native vegetation) sites. Above‐ and belowground rates of ant predation were estimated at these same sites using mealworms, Tenebrio molitor L. (Coleoptera: Tenebrionidae), as baits, simulating crop herbivores. Belowground predation rates were significantly greater in the non‐crop sites, despite the lack of difference in overall abundance and species richness of ants foraging belowground between the crop vs. non‐crop sites. In contrast, we did not detect any significant difference in aboveground predation rates between crop vs. non‐crop sites even though there were significantly more species of ants foraging aboveground in the non‐crop sites. Army ants (subfamily Dorylinae) were the main predatory species belowground, and their abundance was significantly greater in non‐crop sites. In contrast, the main predators aboveground were omnivore ants of the genera Pheidole and Solenopsis, which had similar abundances in the crop and non‐crop sites. Overall, our results indicate that transformation of native Cerrado habitats into crop plantations reduces the abundance of some important predatory species, notably those that forage belowground, and this may negatively affect the potential for ants to provide pest control services in agroecosystems.     

Diploidization and genome size change in allopolyploids is associated with differential dynamics of low‐ and high‐copy sequences

Recent advances have highlighted the ubiquity of whole‐genome duplication (polyploidy) in angiosperms, although subsequent genome size change and diploidization (returning to a diploid‐like condition) are poorly understood. An excellent system to assess these processes is provided by Nicotiana section Repandae, which arose via allopolyploidy (approximately 5 million years ago) involving relatives of Nicotiana sylvestris and Nicotiana obtusifolia. Subsequent speciation in Repandae has resulted in allotetraploids with divergent genome sizes, including Nicotiana repanda and Nicotiana nudicaulis studied here, which have an estimated 23.6% genome expansion and 19.2% genome contraction from the early polyploid, respectively. Graph‐based clustering of next‐generation sequence data enabled assessment of the global genome composition of these allotetraploids and their diploid progenitors. Unexpectedly, in both allotetraploids, over 85% of sequence clusters (repetitive DNA families) had a lower abundance than predicted from their diploid relatives; a trend seen particularly in low‐copy repeats. The loss of high‐copy sequences predominantly accounts for the genome downsizing in N. nudicaulis. In contrast, N. repanda shows expansion of clusters already inherited in high copy number (mostly chromovirus‐like Ty3/Gypsy retroelements and some low‐complexity sequences), leading to much of the genome upsizing predicted. We suggest that the differential dynamics of low‐ and high‐copy sequences reveal two genomic processes that occur subsequent to allopolyploidy. The loss of low‐copy sequences, common to both allopolyploids, may reflect genome diploidization, a process that also involves loss of duplicate copies of genes and upstream regulators. In contrast, genome size divergence between allopolyploids is manifested through differential accumulation and/or deletion of high‐copy‐number sequences.     

Evaluating Drosophila suzukii immunomarking for mark‐capture research

Drosophila suzukii Matsumura (Diptera: Drosophilidae) utilizes ‘Himalaya’ blackberry, Rubus armeniacus Focke (Rosaceae), as a host and may invade berry and stone fruit crops from field margins containing this invasive weed. Laboratory and semi‐field studies were conducted to determine (1) the persistence of protein marks including 10% chicken egg whites (egg albumin protein), 20% bovine milk (milk casein protein), and 20% soy milk (soy trypsin inhibitor protein) on topically sprayed D. suzukii, (2) protein retention on blackberry leaves, and (3) D. suzukii acquisition of protein after exposure to marked blackberry leaves for up to 14 days after application. All flies and leaves were assayed for the presence of the protein marks using protein‐specific enzyme‐linked immunosorbent assays. Egg albumin, milk casein, and soy trypsin proteins persisted on 94, 49, and 25% of the topically marked D. suzukii, respectively, throughout the 14‐day study period. Egg albumin was retained on 100% of treated leaves for 14 days, regardless of environmental conditions. At least 50% of flies exposed residually to egg albumin‐treated leaves were marked for 3 days, regardless of exposure time and environmental conditions. However, increasing fly exposure time to treated leaves in April and June appeared to improve protein mark acquisition. Acquisition of protein by flies from treated leaves for milk casein was inconsistent, and poor for soy trypsin, despite detectable levels on treated leaves. Egg albumin had the longest and most consistent persistence on flies, leaves, and flies exposed to leaves in laboratory and semi‐field studies, under a variety of environmental conditions and exposure times.     

Dietary specialization in mutualistic acacia‐ants affects relative abundance but not identity of host‐associated bacteria

Acacia‐ant mutualists in the genus Pseudomyrmex nest obligately in acacia plants and, as we show through stable isotope analysis, feed at a remarkably low trophic level. Insects with diets such as these sometimes depend on bacterial symbionts for nutritional enrichment. We, therefore, examine the bacterial communities associated with acacia‐ants in order to determine whether they host bacterial partners likely to contribute to their nutrition. Despite large differences in trophic position, acacia‐ants and related species with generalized diets do not host distinct bacterial taxa. However, we find that a small number of previously undescribed bacterial taxa do differ in relative abundance between acacia‐ants and generalists, including several Acetobacteraceae and Nocardiaceae lineages related to common insect associates. Comparisons with an herbivorous generalist, a parasite that feeds on acacias and a mutualistic species with a generalized diet show that trophic level is likely responsible for these small differences in bacterial community structure. While we did not experimentally test for a nutritional benefit to hosts of these bacterial lineages, metagenomic analysis reveals a Bartonella relative with an intact nitrogen‐recycling pathway widespread across Pseudomyrmex mutualists and generalists. This taxon may be contributing to nitrogen enrichment of its ant hosts through urease activity and, concordant with an obligately host‐associated lifestyle, appears to be experiencing genomewide relaxed selection. The lack of distinctiveness in bacterial communities across trophic level in this group of ants shows a remarkable ability to adjust to varied diets, possibly with assistance from these diverse ant‐specific bacterial lineages.     

Species‐Specific PCR‐Based Assay for Identification and Detection of Phomopsis (Diaporthe) azadirachtae Causing Die‐Back Disease in Azadirachta indica

Die‐back disease caused by Phomopsis (Diaporthe) azadirachtae is the devastating disease of Azadirachta indica. Accurate identification of P. azadirachtae is always problematic due to morphological plasticity and delayed appearance of conidia. A species‐specific PCR‐based assay was developed for rapid and reliable identification of P. azadirachtae by designing a species‐specific primer‐targeting ITS region of P. azadirachtae isolates. The assay was validated with DNA isolated from different Phomopsis species and other fungal isolates. The PCR assay amplified 313‐bp product from all the isolates of P. azadirachtae and not from any other Phomopsis species or any genera indicating its specificity. The assay successfully detected the pathogen DNA in naturally and artificially infected neem seeds and twigs indicating its applicability in seed quarantine and seed health testing. The sensitivity of the assay was 100 fg when genomic DNA of all isolates was analysed. The PCR‐based assay was 92% effective in comparison with seed plating technique in detecting the pathogen. This is the first report on the development of species‐specific PCR assay for identification and detection of P. azadirachtae. Thus, PCR‐based assay developed is very specific, rapid, confirmatory and sensitive tool for detection of pathogen P. azadirachtae at early stages.     

Cannibalism of parasitoid‐attacked conspecifics in a non‐carnivorous caterpillar

Cannibalism, the killing and consumption of conspecifics, can even occur in insect species typically considered to be non‐carnivorous. Of particular interest is the cannibalism of parasitoid‐attacked conspecifics, which could reduce parasitism levels in subsequent generations for that conspecific population. This study reports on the occurrence and some of the consequences of cannibalism in parasitoid‐attacked obliquebanded leafroller, Choristoneura rosaceana (Harris) (Lepidoptera: Tortricidae). We show that larvae of C. rosaceana, which is considered to be an herbivorous caterpillar species, did not prey upon live conspecifics, but readily consumed conspecifics attacked by Habrobracon gelechiae Ashmead (Hymenoptera: Braconidae). Further examination found that C. rosaceana larvae feeding on parasitoid‐attacked conspecifics, since their fourth instar, suffered a higher mortality and reduction in body size than those fed on plant material only. The cannibalism of attacked conspecifics did not appear to offer any nutrient benefits for the cannibal. To our best knowledge, this is the first empirical example of the occurrence and some of the consequences of cannibalism by a non‐carnivorous insect on its parasitoid‐attacked conspecifics. We discuss the adaptive significance of such cannibalism on parasitoid‐attacked conspecifics with respect to a trans‐generational fitness gain for the population through the killing of the parasitoids, thereby reducing parasitism in subsequent generations.     

Syntaxin 4 mediates endosome recycling for lytic granule exocytosis in cytotoxic T‐lymphocytes

Adaptive and innate immunity utilize the perforin‐killing pathway to eliminate virus‐infected or cancer cells. Cytotoxic T‐lymphocytes (CTLs) and natural killer cells mediate this process by releasing toxic proteins at the contact area with target cells known as immunological synapse (IS). Formation of a stable IS and exocytosis of toxic proteins requires persistent fusion of Rab11a recycling endosomes with the plasma membrane (PM) that may assure the delivery of key effector proteins. Despite the importance of the recycling endosomal compartment, the membrane fusion proteins that control this process at the IS remain elusive. Here, by performing knockdown experiments we found that syntaxin 4 (STX4) is necessary for cytotoxic activity and CD107a degranulation against target cells in a similar fashion to syntaxin 11, which is involved in lytic granule (LG) exocytosis and immunodeficiency when it is mutated. Using total internal reflection fluorescent microscopy we identified that STX4 mediates fusion of EGFP‐Rab11a vesicles at the IS. Immunoprecipitation experiments in lysates of activated CTLs indicate that endogenous STX4 may drive this fusion step by interacting with cognate proteins: Munc18‐3/SNAP23/VAMP7 and/or VAMP8. These results reveal the role of STX4 in mediating fusion of Rab11a endosomes upstream of lytic granules (LGs) exocytosis and further demonstrate the importance of this pathway in controlling CTL‐mediated cytotoxicity.      

Diet type modifies ingestion rates and trophallactic exchanges in leaf‐cutting ants

Conventional control of leaf‐cutting ants is mainly carried out by application of solid insecticide baits. However, alternative approaches could increase the efficiency of control methods. Here, we investigated the hypothesis that liquid and semi‐solid baits are more readily ingested by leaf‐cutting ants and are therefore more toxic than solid baits. Furthermore, following increased ingestion rates, ants could perform higher frequencies of trophallactic exchanges with their nest mates, thus increasing dispersal rates within the colony. Each of three diets were offered to Acromyrmex subterraneus subterraneus (Forel) and Atta sexdens rubropilosa Forel (both Hymenoptera: Formicidae) for 1 h under laboratory conditions and subsequently crop volumes were quantified. The highest crop volumes for both species were seen when the diet was offered in liquid form, and the lowest crop volumes were observed after offering solid diets. Survival rates of the ants were monitored following addition of the neonicotinoid insecticide imidacloprid (IMI) to the three diets. The ingestion of liquid and semi‐solid diets + IMI by A. subterraneus resulted in 17 and 6% survival, respectively, whereas these diets caused 100% mortality of A. sexdens. Ingestion of solid diets with IMI resulted in 51% survival of A. subterraneus and 23% survival of Atta workers. Twenty‐three percent of A. subterraneus which had fed on liquid diet carried out trophallactic exchanges, whereas only 10 and 3% of ants carried out trophallaxis when fed on semi‐solid or solid diet, respectively. Lower trophallactic frequencies were observed for A. sexdens, with 13 and 3% of ants that had fed on liquid and semi‐solid diets performing trophallaxis, respectively. The low trophallactic exchange rates following ingestion of solid diets would reduce the dispersal of these compounds throughout the colony. Control strategies using solid baits should be re‐examined in the light of these findings, which indicate the importance of optimizing insecticide ingestion.     

Artificial selection,pre‐release diet,and gut symbiont inoculation effects on sterile male longevity for area‐wide fruit‐fly management

Longevity is an important life‐history trait for successful and cost‐effective application of the sterile insect technique. Furthermore, it has been shown that females of some species – e.g., Anastrepha ludens (Loew) (Diptera: Tephritidae) – preferentially copulate with ‘old’, sexually experienced males, rather than younger and inexperienced males. Long‐lived sterile males may therefore have greater opportunity to find and mate with wild females than short‐lived males, and be more effective in inducing sterility into wild populations. We explored the feasibility of increasing sterile male lifespan through selection of long‐lived strains and provision of pre‐release diets with added protein, and inoculated with bacterial symbionts recovered from cultures of the gut of wild Anastrepha obliqua (Macquart). Artificial selection for long‐lived A. ludens resulted in a sharp drop of fecundity levels for F1 females. Nevertheless, the cross of long‐lived males with laboratory females produced a female F1 progeny with fecundity levels comparable to those of females in the established colony. However, the male progeny of long‐lived males*laboratory females did not survive in higher proportions than laboratory males. Provision of sugar to A. obliqua adults resulted in increased survival in comparison to adults provided only with water, whereas the addition of protein to sugar‐only diets had no additional effect on longevity. Non‐irradiated males lived longer than irradiated males, and supplying a generic probiotic diet produced no noticeable effect in restoring irradiated male longevity of A. obliqua. We discuss the need to evaluate the time to reach sexual maturity and survival under stress for long‐lived strains, and the inclusion of low amounts of protein and specific beneficial bacteria in pre‐release diets to increase sterile male performance and longevity in the field.