The biology of Laricobius nigrinus Fender (Coleoptera: Derodontidae) and its potential for control of hemlock woolly adelgid (HWA), Adelges tsugae Annand (Homoptera: Adelgidae) in the eastern United States were investigated. Laricobius nigrinus completed development on HWA in laboratory studies. There are four larval instars. Mean larval consumption was 225.9 and 252.3 HWA eggs at 12 and 18 °C, respectively. Post-aestivation activity period was 36.6 and 30.8 weeks for males and females at 13 °C, respectively. Mean lifetime fecundity was 100.8 eggs over a mean ovipositional period of 13.2 weeks. Laricobius nigrinus is host specific. In paired-choice and no-choice oviposition tests, L. nigrinus laid more eggs in HWA ovisacs than other test species. Laricobius nigrinus consumed more eggs of HWA than eggs of Adelges piceae (Ratzeburg) and Pineus strobi (Hartig) but not of Adelges abietis (L.). In larval development tests, L. nigrinus only completed development on HWA. A two-year field study conducted in British Columbia showed that: 1) the life cycle of HWA in British Columbia is similar to that previously reported in Virginia and Connecticut; 2) L. nigrinus adults undergo an aestival diapause that coincides with the diapause of the first instar HWA sistens. Adult activity beginning in autumn coincides with resumption of development of the sistens generation; 3) oviposition and subsequent larval development of L. nigrinus coincide with oviposition by HWA sistens adults. Temperature-dependent development was determined for egg, larval, pre-pupal and pupal stages of L. nigrinus at five constant temperatures (9, 12, 15, 18 and 21 °C). Development time was inversely proportional to temperature between 9 and 18 °C. Laricobius nigrinus did not complete development at 21 °C. Minimum developmental temperatures for eggs (6.5 °C), larvae (5.1 °C), pre-pupae (3.8 °C), and pupae (3.1 °C) were determined by extrapolation of linear regression equations to the x-intercept. Degree-days calculated for eggs, larvae, pre-pupae, pupae, and egg to adult were 54.6, 161.3, 196.1, 212.8, and 666.7, respectively. The laboratory derived degree-day model for predicting egg hatch was validated with field data collected in British Columbia. Observed median egg hatch at three of four sites over two years was within 15% of the predicted DD value (54.6). / Ph. D.

Hemlock woolly adelgid (HWA), Adelges tsugae Annand (Hemiptera: Adelgidae), is an introduced pest injurious to hemlock trees (Tsuga spp.) in the eastern United States. HWA currently infests hemlock in over 50% of its geographic range and has the potential to spread throughout its entire range. Since HWA populations in the eastern United States are not regulated effectively by natural enemies (Wallace and Hain 2000), classical biological control is the most promising option for controlling this pest in the forest setting. This work evaluates Laricobius nigrinus Fender (Coleoptera: Derodontidae), a predator associated with HWA in western North America, is being evaluated as a potential biological control agent for HWA (Cheah et al. 2004). Observations suggest that natural enemies may help keep HWA populations below injurious levels in the western United States (Mausel, pers. comm.). Laboratory studies have revealed that L. nigrinus feed selectively on HWA and field studies in British Columbia, Canada have shown that its lifecycle is highly synchronous with HWA phenology. This predator is being further evaluated for its suitability as a biological control agent. In order to be a viable candidate, L. nigrinus must survive under natural field conditions in the eastern United States, reduce the density of HWA, and it must be amenable to mass rearing. The survival, reproductive capability and predator efficiency of L. nigrinus were assessed over 3 years in the field. In addition, the environmental factors regulating processes in the life cycle to develop and improve rearing procedures for this insect were identified. In the first year, adults survived from February - May, laid up to 41 eggs/beetle, and consumed approximately 4.3 adelgids per day. In the second season, L. nigrinus adults survived from November - April, laid up to 38 eggs/beetle and consumed approximately 4.5 adelgids per beetle throughout the study. In both seasons, adelgid populations were significantly lower on branches with predators than those without predators. This impact was demonstrated on both the winter and spring generation of HWA. The feasibility of caged field releases of L. nigrinus was determined in a third field study. An estimated 10, 000 L. nigrinus eggs were liberated in field cages in spring 2003. The density of adelgids in the subsequent (spring) generation of the adelgids was significantly lower on branches with larval activity than those with no predators. Despite extensive sampling, no F1 adults were observed, however F2 adults were recovered in the fall of 2004, 20 months after release. The L. nigrinus life stages incurring high mortality during rearing were identified and factors affecting survival in the feeding and non-feeding life stages were investigated. Studies on the survival and feeding of adults, length of ovipositional period, density per cage, and survival of larvae were conducted. The effect of type of pupation medium, moisture level, disturbance, soil sterilization, temperature, and photoperiod on survival of the non-feeding stages and time of emergence from aestivation was examined. The most noteworthy finding, regarding rearing, is that aestivation can be extended by storing adults at high temperatures and long daylength throughout the summer and decreasing the temperature and daylength in the fall. Based on the results of these studies, procedures for rearing L. nigrinus have been developed and it is currently being reared at two other institutions. To date, over 8, 000 adults produced at Virginia Tech have been released in 6 states and both F1 and F2 adults have been recovered from multiple locations. / Ph. D.

The biology and host-specificity of <i>Laricobius kangdingensis</i> sp. n. (Coleoptera: Derodontidae), a new predator of hemlock woolly adelgid (HWA), <i>Adelges tsugae</i> Annand (Homoptera: Adelgidae) were investigated and compared with that of a related predator of HWA, <i>L. nigrinus</i> Fender. Adults became active in June 2003; the males emerged first and in greater numbers than the females. The sex ratio of female to male was 1: 1.1. <i>Laricobius kangdingensis</i> sp. n. has four larval instars. The total number of eggs laid between 7 July 2003 and 7 August 2004 was 1327. Mean lifetime fecundity per female was 196.4 ± 53.4 eggs and 97.9 eggs for the P1 (parental gen.) and F1 generations, respectively. In comparison, lifetime fecundity for F1 <i>L. nigrinus</i> was 100.8 ± 89.6 eggs. Developmental rates were studied in fall 2003 and spring 2004. Combined with data gathered previously at 12 and 15ºC in early 2003, the 6, 9, and 18ºC data were used to determine lower temperature development thresholds for the egg (2.8ºC), larval (1.6ºC), and pre-pupal (5.8ºC) stages. The egg and larval thresholds are lower than those observed for <i>L. nigrinus</i>. <i>Laricobius kangdingensis</i> sp. n. individuals completed development only at 12 and 15ºC; however, these adults did not emerge from aestivation after pupation. <i>Laricobius kangdingensis</i> sp. n. appears to be host-specific. In paired-choice feeding tests, adults consumed significantly more HWA than two other adelgid species [eastern spruce gall adelgid, <i>Adelges abietis</i> (L.), and pine bark adelgid, <i>Pineus strobi</i> (Hartig)], the pine needle scale, <i>Chionaspis pinifoliae</i> (Fitch), and the woolly alder aphid, <i>Paraprociphilus tessellatus</i> (Fitch). In paired-choice oviposition tests, females laid more eggs on HWA-infested eastern hemlock than on any other test prey. In no-choice host suitability experiments larvae fed on HWA; eastern spruce gall adelgid; the larch adelgid, <i>A. laricis</i> Vallot; the balsam woolly adelgid, <i>A. piceae</i> (Ratzeburg); pine bark adelgid; and pine needle scale, but not on woolly alder aphid or elongate hemlock scale, <i>Fiorinia externa</i> Ferris. Although no larvae completed development in any of the bioassays, 6 out of 15 larvae (40%) in the HWA treatment reached the pre-pupal stage. One larva out of 6 reached the pre-pupal stage on pine bark adelgid. / Master of Science

Insects form close partnerships, or symbioses, with bacteria. These partnerships allow the insects to use resources that would be unavailable to them otherwise. Certain insects, hemipterans, are able to feed on nutrient-poor plant-sap because these bacteria supplement their diets. While this association is beneficial for both the insect and bacteria, it also comes with consequences: the genomes of bacterial symbionts typically undergo extreme degradation, becoming small and lacking many genes necessary for typical bacterial functioning. In the Hemiptera, aphids, mealybugs, cicadas, true bugs,and their relatives, these bacterial partnerships tend to be stable over millions of years. However, adelgids (Aphidoidea: Adelgidae) are highly unusual in that their symbiotic bacteria have been frequently replaced. These replacements offer a unique opportunity to explore the effects of symbiont role and age on symbiont genome degradation. My dissertation uses the pattern of adelgid symbiont gains and losses to understand the process of symbiont replacements and co-symbiont gain. I accomplished this by sequencing and annotating the genomes of adelgid symbionts from across the family, first focusing on the symbionts from a pest species, then expanding to representatives from across the family, and finally conducting an in-depth exploration of how the genomes of a symbiont found in two branches of the adelgids varies between species. Through this work I demonstrate that adelgid symbionts are nutritional providers, they have a unique pattern for distributing the work of providing nutrients between the symbiont pairs, and that a symbiont’s precedence, whether it was there first or whether it joined another symbiont, has an impact on genome degradation.