危害松树的小蠹虫与其伴生菌的相互关系

危害健康松属植物的小蠹虫经常与一些特殊的真菌相联系。在小蠹虫危害松属植物的过程中,这些真菌被小蠹虫的一些特殊结构或者体表携带到松属植物上。小蠹虫与其伴生菌的联系表明小蠹虫和其伴生菌之间是一种互惠互利的关系。伴生菌随小蠹虫扩散而被带到新的寄主树木。而伴生菌或作为小蠹虫的食物来源,但更重要的是,有些伴生菌能够通过其菌丝渗透寄主组织,释放毒素,致死寄主树木,以帮助小蠹虫降低寄主抗性。许多研究致力于探索小蠹虫/伴生菌联合体与寄主树木之间关系的特征和确定小蠹虫与其伴生菌相互关系在生态学上的意义。然而,不同小蠹虫和其伴生菌所组成的共生体系,不同小蠹虫的种群数量,和不同环境条件下同种小蠹虫与其伴生菌相互作用方式的差异使我们在研究小蠹虫和其伴生菌这个共生体系时,对它们各自在成功聚集寄主树木过程中所发挥的重要作用的概括变得非常困难。     

Mycangia of the ambrosia beetle, Austroplatypus incompertus (Schedl) (Coleoptera: Curculionidae: Platypodinae)

Abstract  Ambrosia beetles have an obligate relationship with the ambrosia fungi that they feed on. This requires that the beetles have means to transport those fungi when they colonise new hosts. Some ambrosia beetles have special structures called mycangia to transport fungi in. This paper describes the mycangia of the ambrosia beetle Austroplatypus incompertus and illustrates how the mycangical hairs are probably used by the beetle to acquire fungal spores for transport. The mycangia and probable method of fungal acquisition of this species are compared with those of other ambrosia beetles.     

Origin and evolution of fungus farming in wood-boring Coleoptera – a palaeontological perspective

Insect–fungus mutualism is one of the better-studied symbiotic interactions in nature. Ambrosia fungi are an ecological assemblage of unrelated fungi that are cultivated by ambrosia beetles in their galleries as obligate food for larvae. Despite recently increased research interest, it remains unclear which ecological factors facilitated the origin of fungus farming, and how it transformed into a symbiotic relationship with obligate dependency. It is clear from phylogenetic analyses that this symbiosis evolved independently many times in several beetle and fungus lineages. However, there is a mismatch between palaeontological and phylogenetic data. Herein we review, for the first time, the ambrosia system from a palaeontological perspective. Although largely ignored, families such as Lymexylidae and Bostrichidae should be included in the list of ambrosia beetles because some of their species cultivate ambrosia fungi. The estimated origin for some groups of ambrosia fungi during the Cretaceous concurs with a known high diversity of Lymexylidae and Bostrichidae at that time. Although potentially older, the greatest radiation of various ambrosia beetle lineages occurred in the weevil subfamilies Scolytinae and Platypodinae during the Eocene. In this review we explore the evolutionary relationship between ambrosia beetles, fungi and their host trees, which is likely to have persisted for longer than previously supposed.     

REPEATED EVOLUTION OF CROP THEFT IN FUNGUS‐FARMING AMBROSIA BEETLES

Ambrosia beetles, dominant wood degraders in the tropics, create tunnels in dead trees and employ gardens of symbiotic fungi to extract nutrients from wood. Specificity of the beetle–fungus relationship has rarely been examined, and simple vertical transmission of a specific fungal cultivar by each beetle species is often assumed in literature. We report repeated evolution of fungal crop stealing, termed mycocleptism, among ambrosia beetles. The mycocleptic species seek brood galleries of other species, and exploit their established fungal gardens by tunneling through the ambient mycelium‐laden wood. Instead of carrying their own fungal sybmbionts, mycocleptae depend on adopting the fungal assemblages of their host species, as shown by an analysis of fungal DNA from beetle galleries. The evidence for widespread horizontal exchange of fungi between beetles challenges the traditional concept of ambrosia fungi as species‐specific symbionts. Fungus stealing appears to be an evolutionarily successful strategy. It evolved independently in several beetle clades, two of which have radiated, and at least one case was accompanied by a loss of the beetles’ fungus‐transporting organs. We demonstrate this using the first robust phylogeny of one of the world's largest group of ambrosia beetles, Xyleborini.     

Ambrosiodmus rubricollis (Eichhoff) (Coleoptera; Curculionidae; Scolytinae) associated with young Tasmanian blue gum trees

Eucalypts are among the most widely planted forest trees in the world, and outside their native Australian range, the main arthropod pests are sap‐sucking insects, defoliators, gall‐making insects and xylophagous beetles. We report on a new association between a polyphagous wood‐boring beetle and Tasmanian blue gum (Eucalyptus globulus Labill.) in Central Portugal. Unidentified wood‐boring insects were found attacking two three‐year‐old E. globulus trees showing signs of decline among otherwise healthy trees in a commercial plantation, in June 2018. Declining trees presented dead twigs and branches, and recently developed epicormic sprouts evident on the trunks. Insects emerging from logs were identified as Ambrosiodmus rubricollis (Eichhoff), a species native to eastern and southern Asia, with the taxonomic identification validated by molecular analysis. To our knowledge, this is the first record of A. rubricollis associated with a eucalypt species worldwide. It is not clear whether the beetles played any significant role on the decline of the trees, but Ambrosiodmus may be potential pests for several tree and shrub species in Europe, as these beetles can transport pathogenic fungi.     

Influence of global warming on forest coleopteran communities with special reference to ambrosia and bark beetles

Forest insect pests are one of the major disturbance factors in forest ecosystems and their outbreaks are expected to be more severe under the influence of global warming. Coleopterans are dominant among forest insects and their ecological functions include general detritivores, dead wood feeders, fungivores, herbivores, live wood feeders and predators. Ambrosia and bark beetles contribute to ecological succession of forests and, therefore, ecological functions of forests can be changed in response to their outbreaks. Mountain pine beetle (MPB) outbreaks are the most dramatic example of changes in the ecological functions of forest due to the outbreak of a forest insect pest altered by global warming. Composition of coleopteran species varies with latitude. However, composition of functional groups is consistent with latitude which indicates that resources available to beetles are consistent. In coleopteran communities, ambrosia and bark beetles can become dominant due to increases of dead or stressed trees due to the warming climate. This can also induce changes in the ecological functions of coleopterans, i.e. selective force to displace trees that have lower ecological fitness due to temperature increase. Therefore, recent increases in the density ambrosia and bark beetles offer a chance to study ecological processes in forests under the influence of global warming.     

THE EVOLUTION OF AGRICULTURE IN BEETLES (CURCULIONIDAE: SCOLYTINAE AND PLATYPODINAE)

Abstract Beetles in the weevil subfamilies Scolytinae and Platypodinae are unusual in that they burrow as adults inside trees for feeding and oviposition. Some of these beetles are known as ambrosia beetles for their obligate mutualisms with asexual fungi—known as ambrosia fungi—that are derived from plant pathogens in the ascomycete group known as the ophiostomatoid fungi. Other beetles in these subfamilies are known as bark beetles and are associated with free‐living, pathogenic ophiostomatoid fungi that facilitate beetle attack of phloem of trees with resin defenses. Using DNA sequences from six genes, including both copies of the nuclear gene encoding enolase, we performed a molecular phylogenetic study of bark and ambrosia beetles across these two subfamilies to establish the rate and direction of changes in life histories and their consequences for diversification. The ambrosia beetle habits have evolved repeatedly and are unreversed. The subfamily Platypodinae is derived from within the Scolytinae, near the tribe Scolytini. Comparison of the molecular branch lengths of ambrosia beetles and ambrosia fungi reveals a strong correlation, which a fungal molecular clock suggests spans 60 to 21 million years. Bark beetles have shifted from ancestral association with conifers to angiosperms and back again several times. Each shift to angiosperms is associated with elevated diversity, whereas the reverse shifts to conifers are associated with lowered diversity. The unusual habit of adult burrowing likely facilitated the diversification of these beetle‐fungus associations, enabling them to use the biomass‐rich resource that trees represent and set the stage for at least one origin of eusociality.     

Host selection patterns in insects breeding in bracket fungi

Abstract.  1. Fungivorous insects are generally viewed as polyphagous, largely because most fungal fruiting bodies constitute an unpredictable resource. To examine the validity of this hypothesis, and degree of phylogenetic relatedness between the preferred hosts of the insects, host selection in the insect fauna of bracket fungi was studied, using data obtained both from the field and the literature.
2. More than half (53%) of the insect species breeding in them appeared to be monophagous.
3. Modern phylogenies explained the host selection patterns better than older classifications, since non-monophagous species of beetles frequently used hosts that are closely related to each other.
4. The hypothesis that polyphagous species use more heavily decayed fruiting bodies than monophagous species was verified for insects breeding in Fomes fomentarius . The results indicate that the chemical composition of the fungi influences host selection.
5. It is suggested that fruiting bodies of bracket fungi differ from most other fungi in that their occurrence is more predictable. Therefore, the primary colonising fungivores generally attack only one host species, or a few hosts that are closely phylogenetically related. Polyphagous species generally colonise fruiting bodies after they have reached a certain stage of decay, thus escaping their chemical defence.     

Past attacks influence host selection by the solitary bark beetle Dendroctonus micans

1. A spatio‐temporal study of host selection and local spread of a solitary bark beetle attacking live spruce Dendroctonus micans (Kugelann) was carried out using a combination of standard statistical methods, geostatistical analyses, and modelling. The study was based on data from three plots (150–300 trees, 0.3–1 ha) from 1978 to 1993. All trees were mapped and successful and abortive bark‐beetle attacks on each tree were counted annually. Because the attacked trees usually survived, temporal attack patterns as well as spatial patterns could be analysed. 2. The distribution of successful insect attacks on the trees was slightly aggregative, indicating some degree of choice rather than totally random establishment. 3. The level of yearly individual attacks per tree was very stable, suggesting that D. micans usually leave the host in which they develop. 4. The attacked trees were distributed randomly in the plots; at the study's spatial scale, the insects dispersed freely throughout the plot (no spatial dependence). 5. On the other hand, time dependence was strong; some trees were attacked repeatedly while others were left untouched. 6. Among a choice of scenarios (random attack, fixed variability in individual host susceptibility, induced host susceptibility following random attack), the best fit was obtained with the model involving induced individual host susceptibility. This type of relation to the host tree contrasts strongly with patterns generally described in host–plant relationships (including gregarious, tree‐killing bark beetles), where local herbivore damage results in induced resistance. 7. These results suggest that the first attacks in a new stand are made at random, that all or most of the beetles emerging from a tree disperse and resample the stand, and that they settle preferentially on trees that were colonised successfully by previous generations.     

Host specificity of ambrosia and bark beetles (Col., Curculionidae: Scolytinae and Platypodinae) in a New Guinea rainforest

Abstract.  1. Bark and ambrosia beetles are crucial for woody biomass decomposition in tropical forests worldwide. Despite that, quantitative data on their host specificity are scarce.
2. Bark and ambrosia beetles (Scolytinae and Platypodinae) were reared from 13 species of tropical trees representing 11 families from all major lineages of dicotyledonous plants. Standardised samples of beetle-infested twigs, branches, trunks, and roots were taken from three individuals of each tree species growing in a lowland tropical rainforest in Papua New Guinea.
3. A total of 81 742 beetles from 74 species were reared, 67 of them identified. Local species richness of bark and ambrosia beetles was estimated at 80–92 species.
4. Ambrosia beetles were broad generalists as 95% of species did not show any preference for a particular host species or clade. Similarity of ambrosia beetle communities from different tree species was not correlated with phylogenetic distances between tree species. Similarity of ambrosia beetle communities from individual conspecific trees was not higher than that from heterospecific trees and different parts of the trees hosted similar ambrosia beetle communities, as only a few species preferred particular tree parts.
5. In contrast, phloeophagous bark beetles showed strict specificity to host plant genus or family. However, this guild was poor in species (12 species) and restricted to only three plant families (Moraceae, Myristicaceae, Sapindaceae).
6. Local diversity of both bark and ambrosia beetles is not driven by the local diversity of trees in tropical forests, since ambrosia beetles display no host specificity and bark beetles are species poor and restricted to a few plant families.     

Conflict between insect conservation and public safety: the case study of a saproxylic beetle (<Emphasis Type="Italic">Osmoderma eremita</Emphasis>) in urban parks

Urban parks can harbour small populations of saproxylic insects of high conservation concern, such as Osmoderma eremita and other rare beetles. These areas often host old trees which have become very uncommon in rural areas where they are threatened by commercial forestry management procedures based on frequent tree cutting. Nevertheless, old trees of urban parks may represent a hazard for public safety and are sometimes cut by management authorities. The aim of this work was to assess the loss of reproductive sites for saproxylic beetles of the Scarabaeidae, Lucanidae and Cerambycidae, when felling plans are adopted according to a Visual Tree Assessment Procedure (VTA), in a Mediterranean urban park. On July–August 2004, 1,247 holm oaks were surveyed within the border of an urban park of Rome (Villa Borghese). The occurrence of saproxylic beetles (i.e. the presence of frass, living insects or their remains) was verified in 66 old holm oaks, 41% of which were doomed to cutting by VTA. Eleven of these trees (41% of the trees doomed to be cut) held fragments of adults and sometimes living larvae of Osmoderma eremita, and four of them (36%) were included in the felling plan. The presence of Osmoderma eremita in tree holes was more frequent in deep cavities. The presence of frass in the cavities was positively associated with tree height and a high degree of damage at the root collar and negatively with the presence of hole-nesting birds.     

Reducing insecticide volume and nontarget effects of ambrosia beetle management in nurseries

Ambrosia beetles (Coleoptera: Curculionidae: Scolytinae) are increasingly important pests of nursery-grown trees because of the arrival of several invasive species. Ambrosia beetles bore into young trees and inoculate them with ambrosia fungus, which interferes with vascular transport resulting in limb or tree death. In spring, when beetles are active, growers make frequent applications of pyrethroid insecticides to susceptible tree species to deter beetles from boring into trees. Applications often are made with airblast sprayers that forcefully release insecticide mist that billows through nursery beds. Our objective was to compare the environmental, nontarget, and economic effects of airblast sprayer applications to applications made with a new dual-nozzle spray wand that makes targeted applications only to tree trunks where beetles attack. Through replicated experiments at commercial nurseries, we found that 5 times more insecticide was released by airblast sprayers than the manual spray wand. The extra insecticide from airblast applications landed on tree canopies, between rows, and left the nursery beds as drift. As a consequence of not spraying tree canopies, 50% more natural enemies and 50% fewer spider mites were captured in nursery beds treated with the manual spray wand than beds treated with the airblast sprayer. Manual applications require 12 times more labor than airblast applications. However, increased need for expensive miticide applications may make manual applications an economically feasible strategy for integrated pest management (IPM) of ambrosia beetles in nurseries.     

Unintentional introductions of microscopic organisms associated with forest insects

International trade and travel are devastating native flora and fauna in many countries through the intentional and/or unintentional introduction of exotic organisms. Pathway control appears to be particularly effective for microscopic organisms such as mites, nematodes, and fungi that are difficult to see with the naked eye. However, taxonomic and ecological information on such organisms is scarce, sometimes causing time lags or failure in eradication programs. Several groups of mites, nematodes, and fungi commonly share a habitat with insects or use them as dispersal agents (phoresy). Some exotic mites and nematodes are introduced simultaneously with exotic insects, sometimes in wood materials. In Japan, mites, nematodes, and fungi have been collected from lucanid beetles introduced as pets from Southeast Asia. While no lethal nematode species have been collected from lucanid beetles, one hemolymph-sucking mite species, inhabiting the sub-elytral space of its native host, is able to easily switch to the Japanese beetle, Dorcus rectus, killing the insect. Yeasts have also been reported on exotic beetles and laboulbeniomycetes have been found on mites associated with the beetles, although their interactions are unknown. Despite the lack of information available about other mites, nematodes, and fungi associated with intentionally and unintentionally introduced forest insects, our analysis of insect species listed by the International Union for Conservation of Nature suggests that unintentional introductions of the microscopic organisms are quite common as a consequence of the existence of symbiotic relationships such as phoresy and parasitism.     

Endophytic fungi in forest trees: are they mutualists?

Forest trees form symbiotic associations with endophytic fungi which live inside healthy tissues as quiescent microthalli. All forest trees in temperate zones host endophytic fungi. The species diversity of endophyte communities can be high. Some tree species host more than 100 species in one tissue type, but communities are usually dominated by a few host-specific species. The endophyte communities in angiosperms are frequently dominated by species of Diaporthales and those in gymnosperms by species of Helotiales. Divergence of angiosperms and gymnosperms coincides with the divergence of the Diaporthales and the Helotiales in the late Carboniferous about 300 million years (Ma) ago, indicating that the Diaporthalean and Helotialean ancestors of tree endophytes had been associated, respectively, with angiosperms and gymnosperms since ≥300 Ma. Consequently, dominant tree endophytes have been evolving with their hosts for millions of years. High virulence of such endophytes can be excluded. Some are, however, opportunists and can cause disease after the host has been weakened by some other factor. Mutualism of tree endophytes is often assumed, but evidence is mostly circumstantial. The sheer impossibility of producing endophyte-free control trees impedes proof of mutualism. Some tree endophytes exhibit either a pathogenic or a putatively mutualistic behaviour depending on the situation. The lifestyle (mutualism, commensalism, parasitism) of most tree endophytes is, however, not known. They are just there in the tissue and resume growth at the onset of natural senescence of the host tissue on which they eventually sporulate. Density of colonization of conifer needles by endophytes increases with needle age. It is postulated that the needles die as soon as colonization density reaches a threshold value. Normally, the threshold is not reached before the onset of natural senescence. The threshold value may, however, be reached earlier under some adverse conditions, e.g. lack of light in dense stands. As a consequence, endophytes kill the needles prematurely. Needle endophytes could, thus, be useful to eliminate “parasitic” needle mass, i.e. needles which consume more than they produce.     

Fungal flora associated with Tomicus piniperda L. in an area close to a timber yard in southern Poland

Abstract:  The association between Tomicus piniperda L. (Col., Scolytinae) and fungi was studied in a Pinus sylvestris L. forest in Mielec-Mościska. Fungi were isolated from overwintered adult beetles taken from two stands situated in different distance from timber yard. Two media were used for isolation. The results showed great diversity of fungi associated with T. piniperda : 1895 cultures, representing 64 species, were isolated. Penicillia and Hormonema dematioides were the dominant species, found in 20.2% and 17.8% of all beetles, respectively. A frequently isolated ophiostomatoid fungi was Ophiostoma minus . Qualitative and quantitative differences in the mycobiota composition of this insect between two stands were detected. The highest richness and diversity of fungal species appeared in the samples taken from the location where the trees were heavily damaged by shoot-feeding of T. piniperda . Differences were most clear for the pathogenic O. minus , which was a common fungal associate of the insects in this stand.     

Modelling mating success of saproxylic beetles in relation to search behaviour, population density and substrate abundance

We compared the efficiency of two mate-finding strategies exploited by representatives of the beetle families Cisidae and Anobiidae (genus Dorcatoma) that live inside fruiting bodies of wood-decaying fungi. In the Cisidae both sexes are attracted to host odour, but no pheromones seem to be present (nonpheromone strategy). In the Dorcatoma species only the females are attracted to host odour, but having found a host they attract males with a sexual pheromone (pheromone strategy). With a simulation model, we compared the efficiency of the two strategies at four densities of trees hosting fungal fruiting bodies and at three relative densities of insects. We found only small differences in efficiency between the two strategies at high relative densities of conspecific individuals, regardless of host tree density. The pheromone strategy was relatively more efficient when the relative density of insects or the density of host trees decreased. Thus, species adopting the nonpheromone strategy are probably more sensitive to habitat fragmentation and more likely to decline and go extinct at low population densities (because of Allee effects) than species using the pheromone strategy. Copyright 2003 Published by Elsevier Science Ltd on behalf of The Association for the Study of Animal Behaviour.      

Host switching by an ambrosia beetle fungal mutualist: Mycangial colonization of indigenous beetles by the invasive laurel wilt fungal pathogen

Ambrosia beetles require their fungal symbiotic partner as their cultivated (farmed) food source in tree galleries. While most fungal-beetle partners do not kill the host trees they inhabit, since their introduction (invasion) into the United states around ~2002, the invasive beetle Xyleborus glabratus has vectored its mutualist partner (but plant pathogenic) fungus, Harringtonia lauricola, resulting in the deaths of over 300 million trees. Concerningly, indigenous beetles have been caught bearing H. lauricola. Here, we show colonization of the mycangia of the indigenous X. affinis ambrosia beetle by H. lauricola. Mycangial colonization occurred within 1 h of feeding, with similar levels seen for H. lauricola as found for the native X. affinis-R. arxii fungal partner. Fungal mycangial occupancy was stable over time and after removal of the fungal source, but showed rapid turnover when additional fungal cells were available. Microscopic visualization revealed two pre-oral mycangial pouches of ~100–200 × 25–50 μm/each, with narrow entry channels of 25–50 × 3–10 μm. Fungi within the mycangia underwent a dimorphic transition from filamentous/blastospore growth to yeast-like budding with alterations to membrane structures. These data identify the characteristics of ambrosia beetle mycangial colonization, implicating turnover as a mechanism for host switching of H. lauricola to other ambrosia beetle species.     

Behavioral mechanisms involved in reducing competition in bark beetles

Bark beetles feed and reproduce in the phloem/cambium tissue of trees where severe competition between individuals of the same and different species significantly reduces their reproductive success. In this coevolutionary setting, individuals that can avoid competition whenever possible by means of genetically controlled behavioral mechanisms are naturally selected. Avoidance of intraspecific and interspecific competition is accomplished in part even before landing by olfactory perception of specific pheromones and allomones. These olfactory systems may function at least as well after landing. In several species the uniform distribution of attacks on host trees indicates that individuals avoid competition by only attacking if they are at least a minimum distance from other established attacks. Once beetles are under the bark their tunneling patterns indicate that a behavioral mechanism exists to avoid intersecting nearby galleries of competing individuals. Finally, beetles may choose to re-emerge if the expected chances of successful reproduction by continuing their stay become less than the probabilities of finding another host and successfully reproducing.     

The sudden emergence of pathogenicity in insect-fungus symbioses threatens naive forest ecosystems

Invasive symbioses between wood-boring insects and fungi are emerging as a new and currently uncontrollable threat to forest ecosystems, as well as fruit and timber industries throughout the world. The bark and ambrosia beetles (Curculionidae: Scolytinae and Platypodinae) constitute the large majority of these pests, and are accompanied by a diverse community of fungal symbionts. Increasingly, some invasive symbioses are shifting from non-pathogenic saprotrophy in native ranges to a prolific tree-killing in invaded ranges, and are causing significant damage. In this paper, we review the current understanding of invasive insect-fungus symbioses. We then ask why some symbioses that evolved as non-pathogenic saprotrophs, turn into major tree-killers in non-native regions. We argue that a purely pathology-centred view of the guild is not sufficient for explaining the lethal encounters between exotic symbionts and naive trees. Instead, we propose several testable hypotheses that, if correct, lead to the conclusion that the sudden emergence of pathogenicity is a new evolutionary phenomenon with global biogeographical dynamics. To date, evidence suggests that virulence of the symbioses in invaded ranges is often triggered when several factors coincide: (i) invasion into territories with naive trees, (ii) the ability of the fungus to either overcome resistance of the naive host or trigger a suicidal over-reaction, and (iii) an 'olfactory mismatch' in the insect whereby a subset of live trees is perceived as dead and suitable for colonization. We suggest that individual cases of tree mortality caused by invasive insect-fungus symbionts should no longer be studied separately, but in a global, biogeographically and phylogenetically explicit comparative framework.     

Insect fungal symbionts: A promising source of detoxifying enzymes

Summary Many species of insects cultivate, inoculate, or contain symbiotic fungi. Insects feed on plant materials that contain plant-produced defensive toxins, or are exposed to insecticides or other pesticides when they become economically important pests. Therefore, it is likely that the symbiotic fungi are also exposed to these toxins and may actually contribute to detoxification of these compounds. Fungi associated with bark beetles, ambrosia beetles, termites, leaf-cutting ants, long-horned beetles, wood wasps, and drug store beetles can variously metabolize/detoxify tannins, lignins, terpenes, esters, chlorinated hydrocarbons, and other toxins. The fungi (Attamyces) cultivated by the ants and the yeast (Symbiotaphrina) contained in the cigarette beetle gut appear to have broad-spectrum detoxifying abilities. The present limiting factor for using many of these fungi for large scale detoxification of, for example, contaminated soils or agricultural commodities is their slow growth rate, but conventional strain selection techniques or biotechnological approaches should overcome this problem.Presented at the Symposium on Fungal Detoxification at the 48th Annual Meeting of the Society for Industrial Microbiology, Philadelphia, PA, August 4–9, 1991.