Lethal effect of Goniozus legneri on Cactoblastis cactorum: A potential biocontrol agent for inundative releases.

Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae), the cactus moth, is native to South America with a widespread distribution in Argentina. The larvae consume the interior of Opuntia spp. (Cactaceae) plants. The moth was used as a biocontrol agent against invasive non-native Opuntia spp. in many countries around the world. The cactus moth arrived unintentionally in Florida, USA, expanded its range and threatened Opuntia-based agriculture and natural ecosystems in southern North America. The insect is also a pest of cultivated O. ficus-indica L. in Argentina. An endemic South American parasitoid, Goniozus legneri Gordth (Hymenoptera: Bethylidae), is used in inundative biological control programmes against lepidopteran pests. The goal of this work was to evaluate G. legneri as a biocontrol agent to be used in inundative releases against C. cactorum. Mortality of C. cactorum by G. legneri was assessed at different spatial scales, as well as the interactions with Apanteles opuntiarum Martínez & Berta (Hymenoptera: Braconidae), a common Argentine natural enemy of C. cactorum. The ability of G. legneri to paralyse, parasitise and kill C. cactorum was confirmed. The paralysis inflicted on C. cactorum larvae reduced larval damage to the plants by 85%. Using two parasitoid species increased the mortality of C. cactorum larvae, but it was highly dependent on the order of their arrival. The combined mortality caused by both parasitoids was higher than a single one, in particular when G. legneri arrived first (56 ± 1%), suggesting asymmetric competition due to the preference of G. legneri attacking previously parasitised larvae. Goniozus legneri has potential as an inundative biocontrol agent of C. cactorum, but its interaction with the classical biocontrol agent A. opuntiarum needs to be considered.

Attraction of the Air Potato Leaf Beetle, Lilioceris Cheni, (Coleoptera: Chrysomelidae) to leaf Volatiles of the Air Potato, Dioscorea bulbifera.

Air potato, Dioscorea bulbifera L., is an invasive vine found in the southeastern United States and is native to Asia and Africa. The air potato leaf beetle Lilioceris cheni (Coleoptera: Chrysomelidae), is a host specific biological control agent introduced for D. bulbifera control. In this study, odor cues that control the attraction of L. cheni to D. bulbifera were investigated. The first experiment investigated the response of L. cheni to D. bulbifera leaves versus no leaves in the presence or absence of air flow. The experiment showed a significant response of L. cheni to D. bulbifera leaves in the presence of air flow with leaves placed upwind. When air flow and/or leaves were absent, L. cheni dispersed randomly between the upwind and downwind targets, indicating L. cheni uses volatiles from D. bulbifera in host selection. The second experiment investigated L. cheni response to undamaged, larval-damaged, and adult-damaged plants. Lilioceris cheni showed preference to move towards conspecific damaged plants compared to undamaged plants but did not discriminate between larvae-damaged or adult-damaged plants. The third experiment investigated volatile profiles of damaged D. bulbifera plants using gas chromatography coupled with mass spectroscopy. We found significant differences in volatile profiles between adult and larval damaged plants compared to mechanically damaged and undamaged plants, with increases in 11 volatile compounds. However, larval and adult-damaged volatile profiles did not differ. The information acquired during this study could be used to develop strategies to monitor for L. cheni and improve its biological control program.

Spatial and host related genomic variation in partially sympatric cactophagous moth species.

Surveys of patterns of genetic variation in natural sympatric and allopatric populations of recently diverged species are necessary to understand the processes driving intra- and interspecific diversification. The South American moths Cactoblastis cactorum, Cactoblastis doddi and Cactoblastis bucyrus are specialized in the use of cacti as host plants. These species have partially different geographic ranges and differ in patterns of host plant use. However, there are areas that overlap, particularly, in northwestern Argentina, where they are sympatric. Using a combination of genome-wide SNPs and mitochondrial data we assessed intra and interspecific genetic variation and investigated the relative roles of geography and host plants on genetic divergence. We also searched for genetic footprints of hybridization between species. We identified three well delimited species and detected signs of hybridization in the area of sympatry. Our results supported a hypothetical scenario of allopatric speciation in the generalist C. cactorum and genetic interchange during secondary geographic contact with the pair of specialists C. bucyrus and C. doddi that probably speciated sympatrically. In both cases, adaptation to new host plants probably played an important role in speciation. The results also suggested the interplay of geography and host plant use as drivers of divergence and limiting gene flow at intra and interspecific levels.

Employee work status, mental health, substance use, and career turnover intentions: An examination of restaurant employees during COVID-19.

The COVID-19 pandemic has negatively impacted the restaurant industry and employees in the worst possible way. This empirical study aims to examine the relationships between employees' work status (working, furloughed, or laid-off), mental health (psychological well-being and psychological distress), substance use (drug and alcohol use), and career turnover intentions during the pandemic. Analyzing the responses of 585 restaurant employees using structural equation modelling (SEM), findings revealed that working employees experienced higher levels of psychological distress, drug and alcohol use than furloughed employees. Moreover, psychological distress increased drug and alcohol use, as well as career turnover intentions. Lastly, all employees, regardless of their mental health, increased their substance use and indicated a desire to seek future employment in alternate industries during the pandemic. Practical and theoretical implications are discussed in detail.

Low-oxygen atmospheric treatment improves the performance of irradiation-sterilized male cactus moths used in SIT.

As part of sterile insect technique (SIT) programs, irradiation can effectively induce sterility in insects by damaging genomic DNA. However, irradiation also induces other off-target side effects that reduce the quality and performance of sterilized males. Thus, treatments that reduce off-target effects of irradiation on male performance while maintaining sterility can improve the feasibility and economy of SIT programs. Exposure to ionizing radiation induces the formation of damaging free radicals in biological systems that may reduce sterile male performance. Here, we test whether exposure to an anoxic environment for 1 h before and during irradiation improves male performance, while maintaining sterility in males of the cactus moth, Cactoblastis cactorum (Berg). We show that exposure to 1 h of anoxia increases the moth's antioxidant capacity and that irradiation in anoxia after 1 h of anoxic conditioning decreases irradiation-induced oxidative damage to the moth's lipids and proteins. Anoxia treatment that reduced oxidative damage after irradiation also produced moths with greater flight performance, mating success, and longevity, while maintaining F1 male sterility at acceptable levels for SIT. We conclude that anoxia pretreatment followed by irradiation in anoxia is an efficient way to improve the quality of irradiated moths and perhaps lower the number of moths needed for release SIT moth operations.

Effect of egg production dynamics on the functional response of two parasitoids.

Functional response describes the number of hosts attacked by a parasitoid in relation to host densities and plays an important role by connecting behavioral-level processes with community-level processes. Most functional response studies were carried out using simple experimental designs where the insects were confined to a plain and small arena with different host densities during a fixed period of time. With these designs, other factors that might affect the functional response of parasitoids were not analyzed, such as fecundity, age, and experience. We proposed a series of latent-variables Markovian models that comprised an integrated approach of functional response and egg production models to estimate the realized lifetime reproductive success of parasitoids. As a case study, we used the parasitoids Anagyrus cachamai and A. lapachosus (Hymenoptera: Encyrtidae), two candidate agents for neoclassical biocontrol of the Puerto Rican cactus pest mealybug, Hypogeococcus sp. (Hemiptera: Pseudococcidae). The tested species were assessed according to their physiology and prior experience. We estimated the number of mature eggs after emergence, egg production on the first day, egg production rate, the proportion of eggs resorbed, egg resorption threshold, and egg storage capacity. Anagyrus cachamai and A. lapachosus both presented a type III functional response. However, the two parasitoids behaved differently; for A. cachamai, the number of parasitized hosts decreased with female age and depended on the number of mature eggs that were available for oviposition, whereas A. lapachosus host parasitism increased with female age and was modulated by its daily egg load and previous experience. The methodology presented may have large applicability in pest control, invasive species management, and conservation biology, as it has the potential to increase our understanding of the reproductive biology of a wide variety of species, ultimately leading to improved management strategies.

Genomic and ecological evidence shed light on the recent demographic history of two related invasive insects.

Hypogeococcus pungens is a species complex native to southern South America that is composed of at least five putative species, each one specialized in the use of different host plants. Two of these undescribed species were registered as invasive in Central and North America: Hyp-C is a cactophagous mealybug that became an important pest that threatens endemic cactus species in Puerto Rico, and Hyp-AP feeds on Amaranthaceae and Portulacaceae hosts, but does not produce severe damage to the host plants. We quantified genomic variation and investigated the demographic history of both invasive species by means of coalescent-based simulations using high throughput sequencing data. We also evaluated the incidence of host plant infestation produced by both species and used an ecological niche modeling approach to assess potential distribution under current and future climatic scenarios. Our genetic survey evinced the footprints of strong effective population size reduction and signals of genetic differentiation among populations within each species. Incidence of plant attacks varied between species and among populations within species, with some host plant species preferred over others. Ecological niche modeling suggested that under future climatic scenarios both species would expand their distribution ranges in Puerto Rico. These results provide valuable information for the design of efficient management and control strategies of the Puerto Rican cactus pest and shed light on the evolutionary pathways of biological invasions.

Low-oxygen hormetic conditioning improves field performance of sterile insects by inducing beneficial plasticity.

As part of sterile insect technique (SIT) programs, irradiation can effectively induce sterility in insects by damaging germline genomic DNA. However, irradiation also induces other off-target side effects that reduce the quality and performance of sterilized males, including the formation of damaging free radicals that can reduce sterile male performance. Thus, treatments that reduce off-target effects of irradiation on male performance while maintaining sterility can improve the feasibility and economy of SIT programs. We previously found that inducing a form of rapid, beneficial plasticity with a 1-hr anoxic-conditioning period (physiological conditioning hormesis) prior to and during irradiation improves male field performance in the laboratory while maintaining sterility in males of the cactus moth, Cactoblastis cactorum. Here, we extend this work by testing the extent to which this beneficial plasticity may improve male field performance and longevity in the field. Based on capture rates after a series of mark release-recapture experiments, we found that anoxia-conditioned irradiated moths were active in the field longer than their irradiated counterparts. In addition, anoxia-conditioned moths were captured in traps that were farther away from the release site than unconditioned moths, suggesting greater dispersal. These data confirmed that beneficial plasticity induced by anoxia hormesis prior to irradiation led to lower postirradiation damage and increased flight performance and recapture duration under field conditions. We recommend greater consideration of beneficial plasticity responses in biological control programs and specifically the implementation of anoxia-conditioning treatments applied prior to irradiation in area-wide integrated pest management programs that use SIT.

Influence of competition and intraguild predation between two candidate biocontrol parasitoids on their potential impact against Harrisia cactus mealybug, Hypogeococcus sp. (Hemiptera: Pseudococcidae).

When two or more parasitoid species, particularly candidates for biocontrol, share the same target in the same temporal window, a complex of behaviors can occur among them. We studied the type of interactions (competition and intraguild predation) that existed between the nymphal parasitoids Anagyrus cachamai and A. lapachosus (Hymenoptera: Encyrtidae), two candidate neoclassical biocontrol agents against the Puerto Rican cactus pest mealybug, Hypogeococcus sp. (Hemiptera: Pseudococcidae). The surrogate native congener host in Argentina, the cactus mealybug Hypogeococcus sp., was studied to predict which species should be released; in the case that both should be released, in which order, and their potential impact on host suppression. In the laboratory we conducted experiments where different densities of the host mealybug were exposed to naive females of A. cachamai and A. lapachosus sequentially in both directions. Experiments were analyzed by combining a series of competitive behavioral and functional response models. A fully Bayesian approach was used to select the best explaining models and calculate their parameters. Intraguild predation existed between A. cachamai, the species that had the greatest ability to exploit the resource, and A. lapachosus, the strongest species in the interference competition. The role that intraguild predation played in suppression of Hypogeococcus sp. indicated that a multiple release strategy for the two biocontrol agents would produce better control than a single release; as for the release order, A. lapachosus should be released first.

Ecological Niche Modeling to Calculate Ideal Sites to Introduce a Natural Enemy: The Case of Apanteles opuntiarum (Hymenoptera: Braconidae) to Control Cactoblastis cactorum (Lepidoptera: Pyralidae) in North America.

The cactus moth, Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae), is an invasive species in North America where it threatens Opuntia native populations. The insect is expanding its distribution along the United States Gulf Coast. In the search for alternative strategies to reduce its impact, the introduction of a natural enemy, Apanteles opuntiarum Martínez and Berta (Hymenoptera: Braconidae), is being pursued as a biological control option. To identify promising areas to intentionally introduce A. opuntiarum for the control of C. cactorum, we estimated the overlap of fundamental ecological niches of the two species to predict their common geographic distributions using the BAM diagram. Models were based on native distributional data for both species, 19 bioclimatic variables, and the Maxent algorithm to calculate the environmental suitability of both species in North America. The environmental suitability of C. cactorum in North America was projected from Florida to Texas (United States) along the Gulf coastal areas, reaching Mexico in northern regions. Apanteles opuntiarum environmental suitability showed a substantial similarity with the calculations for C. cactorum in the United States. Intentional introductions of A. opuntiarum in the actual distribution areas of the cactus moth are predicted to be successful; A. opuntiarum will find its host in an environment conducive to its survival and dispersal.

Species complex diversification by host plant use in an herbivorous insect: The source of Puerto Rican cactus mealybug pest and implications for biological control.

Cryptic taxa have often been observed in the form of host-associated species that diverged as the result of adaptation to alternate host plants. Untangling cryptic diversity in species complexes that encompass invasive species is a mandatory task for pest management. Moreover, investigating the evolutionary history of a species complex may help to understand the drivers of their diversification. The mealybug Hypogeococcus pungens was believed to be a polyphagous species from South America and has been reported as a pest devastating native cacti in Puerto Rico, also threatening cactus diversity in the Caribbean and North America. There is neither certainty about the identity of the pest nor the source population from South America. Recent studies pointed to substantial genetic differentiation among local populations, suggesting that H. pungens is a species complex. In this study, we used a combination of genome-wide SNPs and mtDNA variation to investigate species diversity within H. pungens sensu lato to establish host plant ranges of each one of the putative members of the complex, to evaluate whether the pattern of host plant association drove diversification in the species complex, and to determine the source population of the Puerto Rican cactus pest. Our results suggested that H. pungens comprises at least five different species, each one strongly associated with specific host plants. We also established that the Puerto Rican cactus pest derives from southeastern Brazilian mealybugs. This is an important achievement because it will help to design reliable strategies for biological control using natural enemies of the pest from its native range.

Documenting Potential Sunn Hemp (Crotalaria juncea L.) (Fabaceae) Pollinators in Florida.

Sunn hemp, Crotalaria juncea L., is a warm-season legume that can be planted in rotation to cash crops to add nitrogen and organic matter to the soils, for weed growth prevention, and to suppress nematode populations. Sunn hemp flowers also provide nectar and pollen for pollinators and enhance biological control by furnishing habitat for natural enemies. Experiments were conducted in Northern and North Central Florida to evaluate bee populations that visited flowers within mixed plots of sunn hemp and sorghum-sudangrass and plots of two sunn hemp germplasm lines. Collections of bees that visited 'AU Golden' and Tillage Sunn flowers indicated that Xylocopa virginica (L.) (Hymenoptera: Apidae), Xylocopa micans Lepeletier (Hymenoptera: Apidae), Megachile sculpturalis Smith (Hymenoptera: Megachilidae), Megachile mendica (Cresson) (Hymenoptera: Megachilidae), and Megachile georgica Cresson (Hymenoptera: Megachilidae) were present in large numbers in May through July and then again in October. Although Tillage Sunn seeds planted in March flowered in May, percent bloom and number of bee visits were low. Compared with short day sunn hemp cultivars, 'AU Golden' plants produced flowers early in the season to provide food and habitat for pollinators and have the potential to produce an abundant seed crop in Northern and North Central Florida.

Untangling the Hypogeococcus pungens species complex (Hemiptera: Pseudococcidae) for Argentina, Australia, and Puerto Rico based on host plant associations and genetic evidence.

Hypogeococcus pungens, a mealybug native of southern South America, is devastating native cacti in Puerto Rico and threatening cactus diversity in the Caribbean, and potentially in Central and North America. The taxonomic status of H. pungens is controversial since it has been reported feeding not only on Cactaceae but also on other plant families throughout its distribution range. However, in Australia, where the species had been exported from Argentina to control weedy American cacti, it was never found on host plants other than Cactaceae. These conflicting pieces of evidence not only cast doubt on the species identity that invaded Puerto Rico, but also have a negative impact on the search for natural enemies to be used in biological control programs against this pest. Here we present reproductive incompatibility and phylogenetic evidences that give support to the hypothesis that H. pungens is a species complex in which divergence appears to be driven by the host plants. The nuclear EF1α and 18S and the mitochondrial COI genes were used as markers to evaluate the phylogenetic relationships among H. pungens populations collected in Argentina, Australia and Puerto Rico feeding on Cactaceae and/or Amaranthaceae. Additionally, we conducted reciprocal crosses between mealybugs from both hosts. Species delimitation analysis revealed two well-supported putative species within H. pungens, one including mealybugs feeding on Amaranthaceae (H. pungens sensu stricto), and a new undescribed species using Cactaceae as hosts. Additionally, we found asymmetric reproductive incompatibility between these putative species suggesting recent reproductive isolation. The Bayesian species delimitation also suggested that the Australian mealybug population may derive from another undescribed species. Overall, the patterns of genetic differentiation may be interpreted as the result of recent speciation events prompted by host plant shifts. Finally, the finding of a single haplotype in the Puerto Rico population suggests only one invasive event. We still need to identify the geographical origin of the pest in order to enable the use of biological control to reduce the threat to cacti diversity in the Caribbean.

Identification of factors influencing flight performance of field-collected and laboratory-reared, overwintered, and nonoverwintered cactus moths fed with field-collected host plants.

Environmental conditions during egg and larval development may influence the dispersal ability of insect pests, thus requiring seasonal adjustment of control strategies. We studied the longest single flight, total distance flown, and the number of flights initiated by wild Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae) to determine whether the flight performance of overwintered cactus moths with a prolonged feeding phase during development differs from nonoverwintered cactus moths. Pupae of field-collected and laboratory-reared moths were transported together from the United States to Switzerland, and flight mills were used to characterize the flight capacity of 24- to 48-h-old adults during their most active period of the diel cycle. The lack of seasonal variation in flight performance of those moths that developed under controlled environment but were fed with field-collected Opuntia cacti showed that seasonal changes in host plant quality did not affect flight. This consistent flight performance in the mass-reared laboratory population throughout the year is beneficial for sterile insect technique programs, which aim to limit the dispersal of this pest. For field-collected C. cactorum, the larger overwintered females performed similarly to nonoverwintered females, indicating that longer feeding time at lower temperature increases body size but does not influence female flight capacity. Young mated females had a similar flight capacity to unmated ones, suggesting that gravid females may play an important role in invading new habitats. For males, overwintering increased the proportion of long-distance flyers, suggesting that they are well-adapted to locate the more sparsely dispersed females in the spring.

Diel flight pattern and flight performance of Cactoblastis cactorum (Lepidoptera: Pyralidae) measured on a flight mill: influence of age, gender, mating status, and body size.

Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae) is an invasive herbivore that poses a serious risk to Opuntia cacti in North America. Knowledge of the flight behavior of the cactus moth is crucial for a better understanding of natural dispersal, and for both monitoring and control. We used computer-linked flight mills to investigate diel flight activity and flight performance in relation to gender, age, mating status, and body size. Maximal flight activity for both mated and unmated moths occurred during twilight, whereas flight activity was low during photophase. The total distance flown and the number of initiated flights within a diel cycle were higher in both unmated and mated females than in males, but the longest single flight was similar in both genders. These findings suggest that pheromone trap captures of males likely indicate the simultaneous presence of females and that mated females might even be in areas where males are not detected yet. Flight performance heterogeneity was large, with a small portion of the population (both males and females) performing long unbroken flights, whereas the majority made short flights. Females had higher pupal and adult body size and shorter longevity than males. A few individuals, particularly young mated females, flying long distances may be important for active spread of a population and the colonization of new habitats. Implications of this study in the control of the cactus moth by using the sterile insect technique are discussed.