Hybridization was evaluated among three Laricobius spp. involved in the biological control of hemlock woolly adelgid (Adelges tsugae Annand). Following lab mating studies, there was no evidence that Laricobius osakensis Montgomery and Yu could produce hybrid progeny with either Laricobius nigrinus Fender or Laricobius rubidus LeConte. Interaction between L. osakensis and L. nigrinus did not result in a lower production of progeny as a result of fitness costs associated with interspecific mating attempts. Laricobius nigrinus and L. rubidus hybrids were produced in the lab and collected in the field. Hybrid progeny showed very little evidence of decreased fitness. For example, there was no significant difference in the number of days it took for hybrids and pure parental species to develop from egg hatch to the prepupal stage, there was no difference among hybrids and pure parental species in the head capsule widths and larval lengths for the first through third instar, and there was evidence of an F2 generation from field collected specimens. Hybrids produced in the lab had intermediate shaped genital paramere angles compared with parental species, and had elytra coloration similar to that of L. rubidus. Hybrids showed no host preference in the lab, but a preference for Adelges tsugae in the field. Of 12 site factors examined, only the number of years that L. nigrinus was present at the site was found to be associated with percent hybrids. Contamination of the L. osakensis colony with Laricobius naganoensis Leschen resulted in the need to develop molecular methods to differentiate L. osakensis from L. naganoensis. Three restriction enzymes were found that correctly differentiated the two species. / Ph. D.

Predation, egg production and survivorship of Laricobius nigrinus Fender, L. rubidus, LeConte, and L. osakensis Montgomery and Shiyaki (proposed), predators of hemlock woolly adelgid, Adelges tsugae, Annand, were investigated in the laboratory and in the field. In individual assays, L. rubidus oviposited fewer eggs than either L. nigrinus or L. osakensis. In assays containing congeneric or conspecific groups of adult Laricobius, L. osakensis preyed upon the greatest number of ovisacs. When all three species were together, the numbers of ovisacs preyed upon were similar to the mean of all three individual species. Adult predators fed on few eggs and did not exhibit any species preference. The numbers of A. tsugae ovisacs fed upon did not differ significantly by groups of congeneric or conspecific Laricobius larvae. Laricobius adults and larvae had high survival rates throughout all experiments. In the field, predators were enclosed in sleeve cages with both high (> 120 ovisacs) and low (< 90 ovisacs) A. tsugae densities for 1 wk. All branches with caged beetles had significantly greater numbers of ovisacs preyed upon than branches caged without beetles. No differences in predation or egg production were found among the conspecific and congeneric groupings. Predation was uniformly higher at the high prey-density than at the low prey-density. Survivorship among predators did not differ significantly at either prey density. Results from both laboratory and field experiments suggest that these species are able to co-exist and can be in the same location for biological control of A. tsugae. / Master of Science in Life Sciences

The hemlock woolly adelgid (HWA) Adelges tsugae (Annand) (Hemiptera: Adelgidae), has spread throughout most of the range of eastern hemlocks, Tsuga canadensis (L.) and the entire range of Carolina hemlocks, Tsuga caroliniana (Engelman) in eastern North America. Forest managers often implement an integrated pest management (IPM) strategy that combines chemical, silvicultural, and biological control tactics to create a more sustainable and effective approach for managing HWA. Laricobius spp. (Coleoptera: Derodontidae) are the primary biological control agents within IPM scenarios for HWA. Imidacloprid (IMI), the most widely used insecticide for HWA management, is typically applied to the soil below HWA infested trees and breaks down into several metabolites, including two with insecticidal properties; 5-hydroxy imidacloprid (5H-IMI) and imidacloprid-olefin (IMI-OLE). Imidacloprid-olefin is known to be up to 14 times as lethal to HWA as IMI and is produced as imidacloprid is first hydroxylated into 5H-IMI and further metabolized into IMI-OLE. Because Laricobius spp. spends half of its life in aestival diapause in soil below HWA infested trees, there is the potential for these biological control agents to encounter toxic residues within an IPM setting. Imidacloprid and its metabolites are known to cause mortality in Laricobius spp. feeding on HWA from previously treated trees, but the impact of soil applied imidacloprid on their subterranean survivorship has not been studied. Furthermore, there is limited knowledge on the subterranean portion of Laricobius spp. life cycle. The first study determined the depth to which Laricobius spp. burrowed during their subterranean life phase using 3D X-ray microscopy. When held in containers with two differing soil compaction intensities, 0.2 kg/cm2 and 0.5 kg/cm2, Laricobius spp. were found to burrow an average (± SE) of 2.7 ± 1.5 cm and 1.4 ± 1.3 cm deep, respectively. The second study assessed the effect of three different imidacloprid soil treatments; soil injection, soil drench and CoreTect tablets, on the subterranean survivorshiop of Laricobius spp. The soil drench application technique applied in fall 2020 consistently resulted in significantly higher IMI, 5H-IMI, and IMI-OLE residues than the other two treatments over both years of this study (2021 and 2022). The soil injection treatments from both 2020 and 2017 resulted in above average field emergence of beetles in 2021 and 2022, with relatively insignificant residue quantities present. This study suggests that the soil injection treatment method provides a lower level risk of metabolite exposure to Laricobius spp. compared to soil drench and CoreTect tablet imidacloprid application methods. / Master of Science in Life Sciences / In eastern North America, there are two species of hemlock trees whose longevity and unique shade tolerant adaptations are important to the ecosystems within which they are found. Eastern hemlocks range from southeastern Canada to the southern Appalachian Mountains, while Carolina hemlocks are restricted to the southern Appalachians from northern Georgia to Virginia. Since its first formal identification in Richmond, VA in 1951, the hemlock woolly adelgid (HWA) has caused widespread damage and mortality throughout much of the range of eastern hemlocks and the entire range of Carolina hemlocks. HWA consumes nutrients and restricts water flow within the tree, eventually leading to tree mortality. Forest managers typically rely on an integrated approach, that combines chemical applications with the simultaneous release of insects that naturally attack HWA on untreated trees within the same forest setting. Imidacloprid is the most widely used insecticide for HWA management and is typically applied to the soil below HWA infested trees. This insecticide is absorbed through the roots and travels through the tree where HWA feed and die as a result. Laricobius spp., the primary insect consuming HWA on infested trees, spends part of their life in a below ground summer dormancy. Imidacloprid and its secondary compounds are known to cause mortality in Laricobius spp. feeding on HWA from treated trees, but the impact of soil applied imidacloprid on their below ground survivorship has not been studied to date. Furthermore, while extensive studies have taken place to understand the biology of Laricobius spp., there is limited knowledge about their dormant state. In one study, the depth to which Laricobius spp. burrowed during their below ground life phase was determined using 3D X-ray microscopy. In a second study, the effect of three common imidacloprid treatments; soil injection, soil drench, and CoreTect tablets, on below ground survivorship was examined. Laricobius spp. were found to burrow an average (± S.E.) of 2.7 ± 1.5 cm and 1.4 ± 1.3 cm deep in soil compacted at 0.2 kg/cm2 and 0.5 kg/cm2, respectively. Chemical residue quantities were greatest in soil treated with soil drench in fall 2020, and were found to significantly impact Laricobius spp. survival. Overall, survival was the lowest from the soil treated with CoreTect tablets in 2020. Surival was the greatest from soil that received soil injection treatments in both 2020 and 2017 and had relatively insignificant imidacloprid residues present where the insects typically occur in the soil. Findings from this study suggest that imidacloprid application using the soil injection method is a less risky technique for chemical control in an IPM setting when compared to soil drench and CoreTect methods.

The hemlock woolly adelgid (HWA), Adelges tsugae Annand (Hemiptera: Adelgidae) is a small, non-native insect that feeds on hemlock (Tsuga spp.) species native to the eastern United States. One biological control agent, Laricobius osakensis, shows promise due to its voracious feeding on and synchronous life cycle with HWA. This beetle species is originally from Japan and has undergone all the necessary testing while in quarantine, gaining permission for field release in 2010. Field releases began in the fall of 2012 and continued the following fall. Reproduction occurred at the first year's field sites with beetles recovered at both sites the following fall. During the second year of field work, beetle recovery was low due to winter temperatures that caused high HWA mortality. All of the L. osakensis beetles released in the field are first reared in an Insectary on Virginia Tech's campus. To improve health of these beetles during rearing, a series of chemical amendments were added to the water baths where HWA infested cut hemlock branches were placed. HWA samples were taken from each treatment and analyzed for carbon, nitrogen and total carbohydrates and beetles emergence was recorded in relation to prescribed treatment. None of the treatments appeared as the best for improving HWA health, but there were differences between the sampling months within the two years. These differences in month correlate with the biological processes occurring within HWA at these times such as reproduction and preparation for aestivation. No treatment helped increase beetle emergence. / Master of Science in Life Sciences

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

The hemlock woolly adelgid (HWA), Adelges tsugae Annand, (Hemiptera: Adelgidae) is an invasive species in the eastern United States and Canada. This scale-like insect feeds on the xylem ray parenchyma cells of eastern hemlock, Tsuga canadensis (L.) Carrière, (Pinales: Pinaceae) and Carolina hemlock, Tsuga caroliniana Englem., resulting in multiple physical and physiological symptoms, all of which commonly leads to tree mortality within 4 to 10 years. Currently, HWA has spread throughout most of the eastern hemlock range and all of the range of Carolina hemlock and threatens the forest ecosystems they serve. In its introduced range, there are no natural enemies specialized to feed on HWA and suppress HWA populations below damaging levels. In the urban environment and at some public locations, the use of chemicals, such as imidacloprid, have been used to temporarily protect hemlocks, however; this does not provide a permanent solution. In the 1990's a classical biological control program for HWA was created with the goal to suppress HWA populations below damaging levels to reduce tree mortality, and to reduce the negative effects associated with the loss of either hemlock species. Presently, four predators have become the main focus of this program: Laricobius nigrinus Fender, Laricobius osakensis Shiyake and Montgomery (Coleoptera: Derodontidae), Leucotaraxis argenticollis (Zetterstedt), and Leucotaraxis piniperda (Malloch) (Diptera: Chamaemyiidae). Throughout the eastern US, La. nigrinus has established populations and expanded its distribution beyond where it has been released. Although it has successfully established and continues to spread, its effect on eastern hemlock health has been unknown. A 1-year study, carried out in 2021, revealed that La. nigrinus predation on the sistens generation not only reduced the sistens generation, but also had a similar net photosynthetic rate to the negative control on 1-2 year old shoot growth. In addition, significantly more shoots were produced for treatment branches with the highest La. nigrinus density. This also led to a significantly reduced aestivating sistens density compared to the no predator treatment. However, predation was insufficient to reduce HWA populations below the suggested damaging threshold of eastern hemlock (< 4 HWA/cm), suggesting that additional predators would be needed. Recently, the western strain of Le. argenticollis had been released in the eastern US. However, its phenology in the eastern US was unknown. A 2-year study, from 2021-2023, revealed that Le. argenticollis eggs and larvae were present when both generations of HWA adults with eggs and HWA nymphs were present. In addition, Le. argenticollis was capable of completing development in NY and VA, showing promise that this species could establish populations in the eastern US. Timing the adult fly releases into cages at the time when sistens were producing eggs resulted in Le. argenticollis phenology to synchronize well with HWA phenology at both locations, thus providing additional support for its release in the eastern US. Another 2-year study conducted in 2021-2022, at five locations (MD, NC, PA, TN, and VA), sought to determine how the predation of La. nigrinus and Le. piniperda, alone and together, would impact the sistens and progrediens generations, as well as how their predation would affect the aestivating sistens density and new shoot growth production. From this study, La. osakensis was found as the dominant Laricobius species present at the TN site, and similarly to La. nigrinus, reduced intact ovisac density in. This provided a small preview in La. osakensis capabilities and how this predator may be impacting HWA populations at locations where it has established. In order to assess Le. piniperda's impact on the progrediens generation, mesh cages were applied to treatment branches at each site. The study found that Le. piniperda could reproduce at all field sites and that the percentage of disturbed progrediens was significantly higher for treatments with Le. piniperda present at all field sites, except at PA in 2022. However, the affect of both of these predators on the aestivating sistens density and eastern hemlock health was inconclusive. In addition, the percentage of new shoot growth was highly variable among treatments at all field sites, and there was no treatment effect on the mean length of new shoot growth produced at all field sites. The results of this study suggests that in order to fully understand the affects of these predators on HWA populations and on hemlock tree health, longer term studies should be conducted. However, this study does provide some insight on the capabilities of these predators in the short term and supports the continued release and redistribution of these predators. / Doctor of Philosophy / The hemlock woolly adelgid (HWA), is an invasive species in the eastern United States and in Canada. This small scale-like insect was introduced likely on nursery stock material from southern Japan and was first discovered in the eastern US in 1951 in Richmond, Virginia. Two generations of HWA occur each year known as the sistens and progrediens; both reproduce without males. This insect feeds on eastern hemlock and Carolina hemlock, and since its discovery has spread through most of their range. While this insect feeds, it weakens the tree, eventually causing tree death. In eastern North America, eastern hemlock is one of the major tree species in the forest and provides a unique habitat for various mammals, birds, fish, and invertebrate species. To protect this habitat, multiple management strategies have been developed. The two main management strategies that are in use are the use of pesticides and the release of predators of HWA. Pesticides have been useful in the urban environment and in parks and forests, but they only last for a short time and can lead to negative affects to the environment. Since the 1990's, the release of insect predators originally found in the native range of HWA, has occurred with the goal of predator establishment, to lower HWA populations, and to lower hemlock tree death in the eastern US. Four small insect predators have been the main focus of this program: Laricobius nigrinus, Laricobius osakensis, Leucotaraxis argenticollis, and Leucotaraxis piniperda. So far La. nigrinus and La. osakensis have populations at multiple locations throughout the eastern US, and a low number of sites have been recently found for Le. argenticollis and Le. piniperda. Past studies have indicated that La. nigrinus has a significant impact to the sistens generation of HWA, but its affect to eastern hemlock health has been unclear. A 1-yr study conducted in VA in 2021, released La. nigrinus adults into mesh cages, with two different population sizes of beetles, to determine if La. nigrinus predation on HWA populations lead to an indirect effect on eastern hemlock health at the branch level. Results confirmed that La. nigrinus had a significant impact to the sistens generation of HWA. Photosynthesis of branches with La. nigrinus, in June, was similar to the photosynthesis of the negative control branches, which started with a lower number of sistens present on the branch at the beginning of the study. Suggesting that branches with La. nigrinus may be healthier compared to branches without La. nigrinus. Laricobius nigrinus predation also significantly reduced the number of aestivating sistens on new shoot growth, but not below damaging levels (< 4 HWA/cm). This suggested that additional predators may be needed to target the progrediens generation of HWA so that HWA populations do not return to damaging levels after La. nigrinus predation. To learn more about the potential for Le. argenticollis to survive and reproduce in the eastern US, a 2-yr study was conducted in 2021 – 2023, to determine when different life stages of Le. argenticollis could occur in NY and VA. Results revealed that Le. argenticollis was able to reproduce in the mesh cages and complete its life cycle. In addition, Le. argenticollis eggs and larvae were present at the time when HWA adults were producing eggs and when HWA nymphs were present for both generations of HWA. These results provide insight on Le. argenticollis life cycle and shows that Le. argenticollis has a good chance in surviving throughout the eastern US. A 2-yr study at five locations in the eastern US (MD, NC, PA, TN, and VA), tried to combine two predators, La. nigrinus and Le. piniperda, to determine if their combined impact would be sufficient to reduce HWA populations and affect eastern hemlock tree health. Three assessments were conducted each year to determine the impacts of La. nigrinus and Le. piniperda on HWA populations and on eastern hemlock shoot growth production as a representation of tree health. Results confirmed that La. nigrinus significantly reduced the sistens generation at all locations, except at TN. At the TN site, La. osakensis was found to be the main predator of the sistens generation and had a similar affect on the sistens generation as La. nigrinus. Leucotaraxis piniperda was able to reproduce at all locations and also had a significant affect on progrediens ovisacs at four of the locations in 2021 (MD, NC, PA, and VA) and at three of the locations in 2022 (MD, NC, and VA). However, the results on the number of aestivating sistens on new shoot growth and eastern hemlock health were inconclusive. Therefore, it is unclear as to how the presence of both of these predators' affect eastern hemlock health. A long term study may be needed in order to understand the relationship between these predators and their indirect affect to eastern hemlock health.

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.

The predatory beetle, Laricobius nigrinus Fender (Coleoptera: Derodontidae), has been released in the eastern U.S. since 2003 for the management of hemlock woolly adelgid (HWA), Adelges tsugae Annand (Hemiptera: Adelgidae). The establishment of L. nigrinus at release sites in Virginia was assessed in 2017 and 2018. Sampling was performed in both years to determine presence or absence L. nigrinus. Stand level HWA densities, tree health, predator-prey ratios, and Laricobius spp. identifications were also assessed at each site. Laricobius nigrinus established at 82% of sites and was the primary species recovered. HWA densities appeared to decline, and tree health appeared to improve in year two. Predator-prey ratios were lower than those indicated in the native range, however much is left to be understood about the dynamics of this system in its introduced range. A separate two-year study to assess the impact of L. nigrinus on HWA was initiated in 2014 (Phase One) at nine sites in the eastern U.S. Significant predation of HWA sistens ovisacs was demonstrated during this period, therefore, it was continued from 2016-2018 (Phase Two) to provide longer-term evaluations. Predator exclusion cages were used to monitor predator and prey populations. In Phase Two of the study, mean ovisac disturbance rates on no-cage branches were significantly greater than caged branches and were as high as 80%, suggesting that L. nigrinus can have a significant impact on the sistens generation. Microsatellite analysis of Laricobius larvae indicated that L. nigrinus was the primary species recovered at study sites. / Master of Science in Life Sciences / Hemlock woolly adelgid (HWA), an invasive insect native to Asia and western North America, is a significant threat to two native hemlock species in the eastern U.S. Since 2003, a predatory beetle, Laricobius nigrinus, has been released for management of HWA. In biological control programs such as this one, it is important to evaluate the ability of predators to establish and impact prey in areas where they are introduced, as this helps guide future management decisions. As such, a study was conducted to examine the ability of L. nigrinus to establish after being released at 26 locations in Virginia. In those investigations we found that L. nigrinus established at 82% of the sites. Although promising, longer-term studies are needed to understand if the predation of HWA by L. nigrinus helps improve the health of our native hemlocks. A separate two-phase study was carried out from 2014-2016 and 2016-2018 evaluating the impact of L. nigrinus on HWA at nine release sites in the eastern U.S. In Phase Two reported here, cages were used to exclude L. nigrinus on some HWA infested branches while on others, no cage was applied to allow free access to HWA. Comparisons between branches were made to determine the level of predation by L. nigrinus. These studies showed a significant impact by L. nigrinus on the winter generation of HWA with as many as 80% of those insects being attacked on study branches which indicates that this species has potential as an effective predator.

Hemlock woolly adelgid, Adelges tsugae Annand, is an invasive pest threatening eastern (Tsuga canadensis (L.) Carrière) and Carolina hemlock (T. caroliniana Englem.) forests in the eastern US. A new predator, Laricobius osakensis Montgomery and Shiyake, has been found in association with A. tsugae in Japan. Laricobius osakensis was evaluated in a series of pre-release studies to assess its potential as a biological control agent for A. tsugae. Host-range studies indicated that L. osakensis is a specific predator that feeds predominantly and reproduces only on A. tsugae. The functional response "prey consumption changes in response to changes in prey density" was similar for both L. osakensis and Laricobius nigrinus Fender adults. However, L. osakensis had a higher numerical response"changes in oviposition in response to changes in prey density"than L. nigrinus. Laricobius osakensis larvae had a higher functional response than L. nigrinus larvae. Laricobius osakensis\' higher numerical and functional response indicates that this species can potentially be more effective than L. nigrinus. In the evaluation of L. osakensis in sleeve cages in the field from December to April high rates of adult survival, feeding, and reproduction were found. A pair of predators in a cage killed on average five adelgids/day. Peak oviposition occurred in March and April. Larvae from eggs placed in the cages reached maturity in 28-50 days, depending on the season, and only 6.7 % died before reaching maturity. Laricobius osakensis was able to survive, feed, develop, and reproduce in USDA cold-hardiness zones 5b and 6a of southwest Virginia. Behavior of L. osakensis and L. nigrinus was qualitatively similar but varied quantitatively. Laricobius osakensis was more active and had a lower association with T. canadensis. Interactions between species were minimal and not detrimental to either. Intrasexual copulation attempts were observed between males and to a lesser extent between females; however, intrasexual interactions were less frequent than intersexual interactions between the two species. Otherwise activity, including oviposition, was not altered by the presence of the other species. These studies indicate that L. osakensis has the potential to be a valuable addition to the natural enemies complex against A. tsugae. / Ph. D.

The pine bark adelgid, Pineus strobi (Hemiptera: Adelgidae) is a native herbivore of eastern white pine, Pinus strobus (Pinales: Pinaceae), in eastern North America. It is a sessile insect that settles on P. strobus and inserts its stylet bundle to feed on the tree’s phloem. Although P. strobi is not considered a serious pest, it shares its range with the invasive hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae). Predators introduced as biological control agents of A. tsugae interact with P. strobi and its native predators, including Laricobius rubidus LeConte (Coleoptera: Derodontidae). Prior to this study, little work had been done to document the phenology or predators of P. strobi, particularly in its southern range. In the present study, the phenology of P. strobi is reported in southwestern Virginia. Patterns in overwintering population dynamics varied notably from those described from this species’ northern range. The number of annual generations could not be measured due to overlap following two distinct spring generations. Adult body size varied seasonally and was greatest in the spring. Variation between observations from the northern and southern ranges of P. strobi indicate phenological plasticity that informs biological control efforts and offers insight into implication of climatic effects on population dynamics of this and related species. Arthropod predators associated with P. strobi in forests of southwest Virginia were collected during a two-year survey. Morphology and DNA barcoding were used for identification. Species of predators found included: Laricobius rubidus (Coleoptera: Derodontidae), a native adelgid specialist, and two species from the dipteran family Chamaemyiidae, Leucopis piniperda Malloch and L. argenticollis Zetterstedt, which are adelgid specialists. Members of the families Cecidomyiidae, Coccinellidae, Chrysopidae, Hemerobiidae, and Syrphidae were also recovered. Most diverse were the Cecidomyiidae, with 15 different species inferred from their DNA barcodes. Additional work was performed to quantify supercooling points of L. rubidus collected from November – December 2016. These will be compared to those of other Laricobius species in a parallel study. Knowledge of this predator complex is beneficial to describing P. strobi ecology, and also with regard to potential biological control of invasive adelgids in the same region. / Ph. D. / The pine bark adelgid, Pineus strobi (Hemiptera: Adelgidae) is a native herbivore of eastern white pine, Pinus strobus (Pinales: Pinaceae), in eastern North America. It is a sedentary insect that settles on P. strobus bark, branches, and needle bases where it inserts its mouthparts to feed on the tree’s fluid nutrients. Even when P. strobi populations are dense, it does not harm the tree and is not considered a serious pest. It does, however, share its range with the invasive hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae). Predators introduced as biological control agents of A. tsugae also interact with P. strobi and its native predators, including Laricobius rubidus LeConte (Coleoptera: Derodontidae). Prior to the current study, little work had been done to document the natural history or predators of P. strobi, particularly in its southern range, both of which are relevant to the management of other invasive adelgid species found there. In the present study, the natural history of P. strobi is reported in southwestern Virginia. Patterns in the frequencies of various life stages present during the winter varied notably from those described from this species’ northern range. The number of annual generations could not be accurately measured due to the degree of overlap following two distinct spring generations. Adult body size varied seasonally and was greatest in the spring. Variation between observations from the northern and southern ranges of P. strobi indicates that its natural history can vary with the climate, which not only informs biological control efforts, but also offers insight into implication of climatic effects on the population dynamics of this and related species. This study also describes arthropod predators associated with the pine bark adelgid in forests of southwest Virginia found during a two-year survey. Physical characteristics and molecular techniques were used in identification. Species of predators found include: Laricobius rubidus (Coleoptera: Derodontidae), a native adelgid specialist, and two species from the family Chamaemyiidae, Leucopis piniperda Malloch (Diptera: Chamaemyiidae) and L. argenticollis Zetterstedt (Diptera: Chamaemyiidae), which are known adelgid specialists. Members of the families Cecidomyiidae, Coccinellidae, Chrysopidae, Hemerobiidae, and Syrphidae were also recovered. Most diverse were the Cecidomyiidae, with 15 different species genetically identified. Additional work was performed to quantify the bodily freezing points of L. rubidus collected from November – December 2016. These will be compared to those of related biological control agents in a parallel study. Knowledge of this predator complex is beneficial to describing P. strobi ecology, and also with regard to potential biological control of other invasive adelgids in the same region.