Darcy A. Reed

Bagrada hilaris (Hemiptera: Pentatomidae), An Invasive Stink Bug Attacking Cole Crops in the Southwestern United States

An invasive stink bug, Bagrada hilaris (Burmeister) (Hemiptera: Pentatomidae), was discovered in the Western Hemisphere in 2008 near Los Angeles, CA, presumably introduced on container shipments arriving at the Port of Long Beach. In the subsequent 4 years, it has spread throughout southern California, southern areas of Arizona, Nevada, and Utah, southern and west-central New Mexico, and extreme western Texas. We initiated studies on the seasonality and host range of this invasive insect as it adapts to this non-native habitat. We have learned that the bagrada bug has a single population peak between July and October in urban areas dominated by native and introduced weeds, and two population peaks (March-May and September-November) timed with the production of cole crops in agricultural areas. In greenhouse tests, we found 14 plants on which bagrada bug fed heavily, out of 38 agricultural crops and weed plants evaluated. The opportunistic use of host plants other than crucifers, the use of soil oviposition sites, and tolerance of warm climates may contribute to its invasive potential.

Manipulation of fifth-instar host (Manduca sexta) ecdysteroid levels by the parasitoid wasp Cotesia congregata.

Although 5th (last) instar parasitized Manduca sexta larvae undergo developmental arrest and do not wander, they exhibit a small hemolymph ecdysteroid peak (300-400pg/&mgr;l) which begins one day prior to the parasitoids molt to the 3rd (last) instar and concomitant emergence from the host. Ecdysteroids present in this peak were 20-hydroxyecdysone, 20,26-dihydroxyecdysone and one or more very polar ecdysteroids, as well as small amounts of 26-hydroxyecdysone and ecdysone. In parasitized day-1 and -2 5th instars ligated just behind the 1st abdominal proleg, hemolymph ecdysteroid levels increased in both anterior and posterior portions (100-500pg/&mgr;l), while in unparasitized larvae, hormone levels only increased in the anterior portion (100-350pg/&mgr;l). Thus, the ecdysteroid peak observed in host 5th instars was probably produced, at least in part, by the parasitoids. It may serve to promote Cotesia congregatas molt from the second to the third instar and/or to facilitate parasitoid emergence from the host. In parasitized day-1 and -2 5th instars ligated between the last thoracic and 1st abdominal segments, hemolymph ecdysteroid titers reached much higher levels (500-3500pg/&mgr;l) in the anterior portion (no parasitoids present) than in the posterior portion (150-450pg/&mgr;l). Therefore, it appears that the parasitoids regulation of hemolymph ecdysteroid titers occurs at two levels. First, parasitization neutralizes the hosts ability to maintain its normal hemolymph ecdysteroid levels. Second, in a separate action, the parasitoid manipulates the ecdysteroid-producing machinery so that hemolymph levels are maintained at the 200-400pg/&mgr;l characteristic of day 3-4 hosts. This is the first report of a parasitoids ability to interfere with the normal inhibitory mechanisms which prevent prothoracic gland production of ecdysteroid at inappropriate periods of insect growth and development.

Effects of parasitization by Cotesia congregata on the brain-prothoracic gland axis of its host, Manduca sexta

The ability of prothoracic glands (PTGs) from parasitized and unparasitized Manduca sexta 5th-instars to respond to ecdysiotropic extracts prepared from day-5 5th instar brains was compared. An in vitro bioassay revealed that PTGs from parasitized animals were much less responsive to brain PTTH than glands from unparasitized larvae. However, when incubated in Graces medium in the absence of brain extract, glands from day-3 and -4 hosts remained active for a much longer period of time than did those dissected from their unparasitized counterparts. Rather than exhibiting reduced (basal) levels of synthesis after the 3rd hour of incubation, glands from these parasitized larvae continued to synthesize/release ecdysteroid into the medium at relatively high rates. The timing of this enhanced secretory activity is coincident with the ecdysteroid peak that occurs just prior to and during wasp emergence. Following parasite emergence, gland activity decreased, and by the third day after emergence, was reduced to low levels. Results suggest that the requirement for PTTH to stimulate ecdysteroid production has been bypassed, i.e. that the parasite has uncoupled the normal mechanisms that permit brain regulation of PTG activity. The ability of brains from parasitized M. sexta to stimulate PTGs from unparasitized day-2 5th instars was also examined. Dose-response analyses performed for the first 7 days of the 5th instar showed that on a per brain basis ecdysiotropic activity in brains from parasitized and unparasitized animals was similar. However, when differences in brain size were considered, ecdysiotropic activity appeared to be more concentrated in brains from day-7 parasitized larvae than in brains from similarly aged unparasitized larvae. Analysis of the size distribution of the ecdysiotropic activity in brains from parasitized larvae revealed a unique form that was larger than the 29kDa standard. This suggests that parasitization may inhibit neuropeptide processing, particularly during the final stages preceding emergence of the wasps from the host. Thus, both an inhibition of prothoracicotropic hormone processing and the inability to respond to this neurohormone may contribute to the developmental arrest characteristic of parasitized 5th instars.

Biology, Ecology, and Management of an Invasive Stink Bug, Bagrada hilaris, in North America.

The painted bug, Bagrada hilaris, native to eastern and southern Africa and Asia, was detected in California in 2008, and it has spread rapidly throughout several southwestern US states. A polyphagous insect, it is particularly damaging to the billion dollar cole crop industry. B. hilaris frequently causes damage when it migrates to newly planted crops from weedy hosts. Feeding produces circular or star-shaped chlorotic lesions that become necrotic, and infested plants may be distorted. Currently, no reliable sampling methods for B. hilaris exist, nor are there effective natural enemies in the United States. Therefore, management has relied on multiple applications of insecticides and cultural practices such as removal of weedy hosts, destruction of crop residues, timing of planting, and use of transplants. Several pyrethroid and neonicotinoid insecticides are most effective for controlling the insect. Reliable sampling methods and further development of integrated pest management strategies to manage this invasive pest are urgently needed as its range continues to expand.

Diel Activity and Behavior of Bagrada hilaris (Hemiptera: Pentatomidae) on Desert Cole Crops

ABSTRACT Patterns of diel activity and behavior of the Bagrada bug, Bagrada hilaris (Burmeister) (Hemiptera: Pentatomidae), were investigated in a series of field and greenhouse experiments in Yuma, AZ. Adults of B. hilaris were monitored by on-site direct observation on broccoli and cauliflower plants in large-block experimental plots at various intervals throughout consecutive 24-h periods. In the field trials, mean number of B. hilaris adults differed among sampling times, with peak abundance observed consistently between 1300 and 1800 hours. There was a positive correlation between temperature and numbers of B. hilaris adults in all fields, and a negative relationship between humidity and adult numbers in 7 of 12 fields. Significant relationships between temperature and B. hilaris numbers were revealed in regression models for all trials. Slopes were not significantly different among all broccoli blocks or in three of the four cauliflower fields. In greenhouse studies, mating occurred throughout the day but peaked between 1000 and 1600 hours. Females of B. hilaris caused more feeding damage than males on the first true leaf of broccoli, and additional differences in behavior between sexes were observed. Our results suggest that higher temperatures increase field activity and influence the behavior of adult B. hilaris, and temperature is a more reliable indicator to predict the activity of B. hilaris than other environmental parameters measured. The implications of these findings for developing monitoring and management programs for B. hilaris in cruciferous crops are discussed.

Susceptibility of Bagrada hilaris (Hemiptera: Pentatomidae) to insecticides in laboratory and greenhouse bioassays.

ABSTRACT Field-collected nymphs and adults of Bagrada hilaris (Burmeister) (Hemiptera: Penatatomidae) from three locations were evaluated for susceptibility to insecticides representing 10 classes of insecticide chemistry. Although relative susceptibilities differed between leaf-spray and leaf-dip Petri dish bioassays, consistently low LC50 values were determined for chlorpyrifos, bifenthrin, and lambda-cyhalothrin. Fenpropathrin and methomyl had intermediate values. Susceptibility to dinotefuran varied depending on the bioassay, possibly owing to leaf substrates used in the two bioassays. In soil systemic bioassays, the LC50 value of dinotefuran was significantly greater than that of two other neonicotinoids, imidacloprid and thiamethoxam, and the anthranilic diamide, cyantraniliprole. Mortality and feeding damage of B. hilaris and plant growth on insecticide-treated plants in greenhouse trials were consistent with the laboratory bioassays; the best results were seen with bifenthrin, methomyl, and chlorpyrifos. Mortality to the neonicotinoids was not evident; however, feeding damage and plant growth responses on dinotefuran-treated plants damage were similar to the noninfested control. This highlights the apparent antifeedant properties of dinotefuran that may have prevented adults from injuring broccoli plants after exposure to foliar spray residues. Data presented serve as baseline susceptibilities that can be used to monitor for resistance development in field populations of B. hilaris.

INHIBITORY EFFECTS OF PARASITISM BY THE GREGARIOUS ENDOPARASITOID COTESIA CONGREGATA ON HOST TESTICULAR DEVELOPMENT

Cotesia congregata is a gregarious larval endoparasitoid of the tobacco hornworm, Manduca sexta. Parasitized larvae exhibit a variety of physiological and developmental aberrations, the most obvious of which is the induction of developmental arrest characterized by the absence of wandering behavior and suppression of pupation. This arrest appears attributable to continued maintenance of an elevated titer of juvenile hormone and reduced levels of hemolymph juvenile hormone esterase activity. Injection of the wasps polydnavirus into nonparasitized larvae also causes arrest and the larvae eventually form larval-pupal intermediates instead of normal pupae, indicating the virus may be partially responsible. Aside from causing arrested host development, parasitism also inhibits the normal development and differentiation of testes in male host larvae, so that the testes atrophy instead of growing synchronously with other larval tissues. Here we report that parasitism has pronounced disruptive cytological effects on the developing reproductive organs of male hosts, in addition to causing them to atrophy. Parasitism results in a reduction in testicular volume attributable to a reduction in the number of developing germ cells. Microscopy revealed that the structural integrity of the sheaths surrounding the testicular follicles also is disrupted, so that the tissues appear grossly abnormal compared to those of nonparasitized larvae. Intrahemocoelic injection of purified C. congregata polydnavirus in combination with venom into nonparasitized fourth instar larvae, or topical application of 100 μg of methoprene to fourth instar larvae, also alters sheath integrity and reduces the numbers of developing germ cells, but not to the same degree as the pattern observed in truly parasitized hosts. The occurrence of cell death in the male gonad was documented using the vital dyes acridine orange and ethidium bromide. Arch. Insect Biochem. Physiol. 36:95–114, 1997.

Development of a Strategy for Selective Collection of a Parasitoid Attacking One Member of a Large Herbivore Guild

Abstract Selectively collecting a single natural enemy species that parasitizes one member of a guild of herbivores that attack the same host plants can be a challenging problem during development of biological control programs. We present here a successful strategy for the collection of a strain of the egg parasitoid Avetianella longoi Siscaro (Hymenoptera: Encyrtidae), that parasitizes eggs of the longhorned borer Phoracantha recurva Newman (Coleoptera: Cerambycidae). This cerambycid is one member of a large guild of woodborers that simultaneously infest dying and fallen Eucalyptus in Australia, and it has become a major pest of Eucalyptus in many areas of the world where Eucalyptus has been introduced. Adult P. recurva of both sexes were caged on freshly cut Eucalyptus logs, and the resulting egg masses were marked and then left exposed to natural parasitization in the field. Parasitized egg masses were then harvested and held in the laboratory until adult parasitoids emerged. Parasitoids were identified as A. longoi by morphological comparisons with reference specimens, and with molecular markers. This strain of A. longoi readily accepted and had high survival rates in eggs of P. recurva. In contrast, the strain of A. longoi that has been used for biological control of P. semipunctata in California since the 1990s strongly prefers eggs of eucalyptus longhorned borer, Phoracantha semipunctata (F.) (Coleoptera: Cerambycidae), and has relatively low rates of survival to adulthood in eggs of P. recurva. The causes of these behavioral and physiological differences between the two strains are not yet known.

Effects of Temperatures on Immature Development and Survival of the Invasive Stink Bug Bagrada hilaris (Hemiptera: Pentatomidae)

Abstract Bagrada hilaris (Burmeister) (Hemiptera: Pentatomidae) is a non-native stink bug that feeds primarily on cole crops and wild mustards. Its invasion into desert agriculture in California and Arizona presents a conundrum between rapid pest development at warm temperatures and severe damage to cool season crops. In this study, the development and survival of B. hilaris were determined at nine constant temperatures (ranging from 20–42°C) when reared on organically grown broccoli florets. Egg hatching was greatly delayed at 20°C, and first instar nymphs did not survive at this temperature. No eggs hatched at 42°C. The highest survival rates (70.0–86.7%) of B. hilaris were observed at temperatures ranging from 24 to 35°C. The total developmental rate of B. hilaris from egg to adult increased from 0.027 to 0.066/d from 24 to 35°C, and then slightly dropped to 0.064/d at 39°C. Based on the linear model, B. hilaris requires 285.4 degree-days to complete its development. The Briere 1 model predicted the lower and upper temperature thresholds as 16.7 and 42.7°C, respectively.The optimal temperature for development (T Opt) was estimated as 36°C. According to the results, B. hilaris is well adapted to warm conditions, and temperatures of 33–39°C are well suited for B. hilaris development. Information from this study helps explain the rapid range expansion of B. hilaris across the southern United States and will be instrumental in predicting future expansion across the rest of the country and in other parts of the world. The relationship between thermal thresholds and invasion dynamics of this pest are discussed.

Allantoin Crystal Formation in Bagrada hilaris (Burmeister) (Heteroptera: Pentatomidae) Females

Abstract Bagrada hilaris is a polyphagous herbivore reported as an invasive pest in the United States. During the course of dissecting Burmeister hilaris unique crystals were observed in both the midgut and oviducts. Crystals were identified using X-ray diffraction techniques. Both acicular (i.e., needle-like, slender, and/or tapered) and cubic (i.e., cube shaped) crystals were observed in six of 75 individuals examined (8.0%). The crystals were mainly observed in females (6.7%), followed by males (1.3%) with no crystals observed in the minimal number of nymphs examined (0%). Crystals of both types were detected in the midgut and lateral oviducts of the females and midgut in males. The acicular crystals often appeared as distinct bundles when present in the midgut and oviducts. Crystals varied in size with the acicular crystals ranging from 0.12 mm to 0.5 mm in length although the cubic crystals ranged in length from 0.25 mm to over 1.0 mm with widths of ∼0.25 mm. The cubic crystals were identified as allantoin although the acicular crystals were most likely dl-allantoin in combination with halite. While allantoin in a soluble form is often found in insect tissues and excreta; being present as a crystal, especially in such a large form, is curious and raises some interesting questions. More research is warranted to further understand mechanisms associated with such crystal formation in B. hilaris and can lead to a better understanding of the excretory process in this species and the role allantoin plays in the elimination of excess nitrogen.