Population size and growth rate, sex ratio and behaviour in the ant isopod, Platyarthrus hoffmannseggi

Female-biased sex ratios are often associated with small, isolated populations. These conditions are exhibited in populations of Platyarthrus hoflmannseggi (Brandt, 1833), a small, blind woodlouse which lives almost solely in ant nests. This study was undertaken to determine how sex ratio varies with population size in P. hoffmunnseggi and how both of these factors affect population growth rate. To accomplish this, a total of 2155 of these isopods were collected from 20 nests of the ant LusiusJaour (Fabricius, 1781). The majority of the isopod populations were female-biased. The behaviour and survivorship of the isopods in their own and foreign antcolonies were compared and suggest that the free movement of P. hoffmannseggi individuals among ant colonies may be extremely limited. These studies also show how isopods are nutritionally linked to their hosts. Measurement of the woodlice revealed two distinct size/age classes in each population and allowed the change in sex ratio from one generation to the next to be estimated. Female-biased populations produce in future generations relatively more females and fewer males than populations of a similar size with a sex ratio of unity. There was no correlation between the degree of female bias and the rate of population growth. These findings are reviewed in the light of recent theoretical sex ratio models and the possible control of the isopods sex allocation by unusual chromosomal mechanisms and intracellular parasites.     

Operational sex ratio in terrestrial isopods: interaction between potential rate of reproduction and Wolbachia-induced sex ratio distortion

Selfish genetic elements distorting sex ratio are known in several arthropods. By inducing a deficit of males, these sex ratio distorters may modify sexual selection by reversing the sex that competes for a mate. They also have potential to reduce the male proportion to values limiting mating possibilities and therefore limiting population size. Wolbachia endosymbionts are intracytoplasmic, vertically transmitted bacteria that convert genotypic males of terrestrial isopods (woodlice) into functional females. We have tested the impact of these feminizing symbionts on the operational sex ratio (OSR) in three woodlice species. Preliminary experiments consisted in estimating the potential rate of reproduction in males and females, and measuring the dynamics of the onset of reproduction in the wild. These parameters were then combined with population sex ratio to discriminate key factors influencing the OSR. The results suggest that the high potential rate of reproduction of males and the asynchrony in female receptivity both counterbalance female-biased sex ratios. The result is an overall balanced or slightly female-biased OSR. Male deficit can therefore not be considered as a factor strongly limiting reproduction in woodlice. Some females were nevertheless found not mated in the wild at the beginning of the reproductive season, most of them being infected by Wolbachia . This suggests that uninfected females may have an advantage as the first mate. Consequences of these findings on woodlice population dynamics are discussed.     

Female remating and sperm competition patterns in a terrestrial crustacean

In most species, both sexes may mate with more than one partner during their life. In terrestrial isopods (woodlice) female remating can occur within a reproductive season (immediate remating) or after a period of sexual rest and sperm storage, that is in a subsequent reproductive season (delayed remating). The pattern of sperm precedence is unknown in both cases. These two female remating patterns may shape male-male competition in different ways. To elucidate both patterns of female remating and sperm precedence, we used an albinism mutation in Armadillidium vulgare to track paternity in laboratory experiments. Males had low remating success after immediate remating attempts, mainly because of the female's refractory behaviour. However, refractory behaviour seemed to be lost after female sexual rest: delayed remating attempts were as successful as first mating attempts with virgin females. In both immediate and delayed remating, competing males had similar fertilization success, but varied in sperm precedence. We hypothesize that males might induce the refractory mating behaviour in females to ensure their paternity. This could be a strategy that evolved in woodlice after the loss of precopulatory mate guarding during adaptation to the terrestrial environment. We discuss the consequences of these findings for woodlice optimal mating strategies. Copyright 2002 The Association for the Study of Animal Behaviour. Published by Elsevier Science Ltd. All rights reserved.     

Effects of thermal and oxygen conditions during development on cell size in the common rough woodlice Porcellio scaber

During development, cells may adjust their size to balance between the tissue metabolic demand and the oxygen and resource supply: Small cells may effectively absorb oxygen and nutrients, but the relatively large area of the plasma membrane requires costly maintenance. Consequently, warm and hypoxic environments should favor ectotherms with small cells to meet increased metabolic demand by oxygen supply. To test these predictions, we compared cell size (hindgut epithelium, hepatopancreas B cells, ommatidia) in common rough woodlice (Porcellio scaber) that were developed under four developmental conditions designated by two temperatures (15 or 22°C) and two air O₂ concentrations (10% or 22%). To test whether small‐cell woodlice cope better under increased metabolic demand, the CO₂ production of each woodlouse was measured under cold, normoxic conditions and under warm, hypoxic conditions, and the magnitude of metabolic increase (MMI) was calculated. Cell sizes were highly intercorrelated, indicative of organism‐wide mechanisms of cell cycle control. Cell size differences among woodlice were largely linked with body size changes (larger cells in larger woodlice) and to a lesser degree with oxygen conditions (development of smaller cells under hypoxia), but not with temperature. Developmental conditions did not affect MMI, and contrary to predictions, large woodlice with large cells showed higher MMI than small woodlice with small cells. We also observed complex patterns of sexual difference in the size of hepatopancreatic cells and the size and number of ommatidia, which are indicative of sex differences in reproductive biology. We conclude that existing theories about the adaptiveness of cell size do not satisfactorily explain the patterns in cell size and metabolic performance observed here in P. scaber. Thus, future studies addressing physiological effects of cell size variance should simultaneously consider different organismal elements that can be involved in sustaining the metabolic demands of tissue, such as the characteristics of gas‐exchange organs and O₂‐binding proteins.     

Ontogenetic shifts in habitat use and activity in a stream-dwelling isopod

We examined ontogenetic changes in diel patterns of habitat use and activity of the lotic isopod Lirceus fontinalis in relation to differences in predation risk from green sunfish, Lepomis cyanellus. Juveniles are ≤5 mm total length (TL), individuals 6-9 mm TL are mature females (c. 90%) and immature males, and mature males are ≥10 mm TL. Isopods of all sizes were far more dense in Cladophora , a filamentous green algae, than in open silt/sand substrates in fish pools. Densities of each size did not increase in open areas at night in fish pools when risk of predation presumably decreases. Few individuals of any size class occurred in open silt/sand areas in fish pools. However, habitat use did not depend on fish presence. Densities and relative abundances of all sizes were similar between algal and open habitats in fishless pools both day and night, except for mature males which were more abundant in open areas during daylight. All sizes of isopods were significantly more dense in fishless versus fish pools, and density of a size class was influenced by fish presence. Densities of size classes were typically juveniles > individuals 6-9 mm TL > mature males. Green sunfish are size selective feeders on isopods, thus demography of isopods within pools may be partially influenced by the size structure of the fish population.
Isopods reduced their activity, as measured by captures in benthic traps, in the presence of caged green sunfish only at night. This behavior coupled with relatively high activity levels during daylight regardless of fish presence suggests other predators or factors may influence diel activity. Mature males were overrepresented in traps relative to their occurrence in fish runs, indicating high activity levels. Mature males were also more active than gravid and nongravid females and juveniles in the presence of fish in laboratory aquaria. Number of male encounters with nongravid females depended on activity level.     

Aggregation in woodlice: social interaction and density effects

Terrestrial isopods are known to be sensitive to humidity, brightness or temperature. Until now, aggregation was assumed to depend on these sensitivities as a result of individual preferences. In this paper, we show that the social component is also important in the isopod aggregation phenomenon. In experimental arenas with two identical shelters up to nearly 90% of woodlice aggregated under shelters. This aggregation was quick as in 10 minutes most of the animals aggregated, irrespective of their density. Nonetheless, 10-15% of the animals walked around the arena, rarely forming very small and short-lasting aggregates outside shelters. Woodlice aggregated preferably under one of the shelters in 77% of experiments. Indeed, almost 80% of the animals out of 40, 60 or 80 animals in the arena aggregated under one shelter. In arenas with 100 individuals the aggregations were proportionally smaller (70%). Our results revealed that 70 animals was a maximum number of woodlice in an aggregate. We concluded that the location of aggregates is strongly governed by individual preferences but the dynamics of aggregation and collective choice are controlled by social interaction between congeners. The tested densities of the animals in the arena did not impact the aggregation patterns.     

Male mate choice based on ontogenetic colour changes of females in the damselfly <Emphasis Type="Italic">Ischnura senegalensis</Emphasis>

While male mate choice behaviour has been reported in many taxa, little is known about its plasticity and evolutionary consequences. In the damselfly Ischnura senegalensis, females exhibit colour dimorphism (gynomorph and andromorph). The body colour of gynomorphs changed ontogenetically in accordance with sexual maturation, while little change occurred in andromorphs. To test the male mate choice between sexually immature and mature females of both morphs, binary choice experiments were conducted. Virgin males that were reared separately from females after emergence did not show significant preference between sexually immature and mature females for both morphs, indicating that virgin males were unable to discriminate female reproductive status. On the other hand, males that had experienced copulation with gynomorphs preferred sexually mature gynomorphs to sexually immature ones. However, males that had experienced copulation with andromorphs could not discriminate between sexually immature and mature andromorphs, probably due to the absence of significant ontogenetic change in their thoracic colour. Therefore, female body colour is an important cue for males in discriminating between sexual maturation stages. Learned mate discrimination depending on copulation experience might help males to detect potential mates effectively and avoid sexually unreceptive immature female. We finally discuss the adaptive significance of the ontogenetic colour change in females.     

Sex-specific plasticity of growth and maturation size in a spider: implications for sexual size dimorphism

Sex-specific plasticity in body size has been recently proposed to cause intraspecific patterns of variation in sexual size dimorphism (SSD). We reared juvenile male and female Mediterranean tarantulas (Lycosa tarantula) under two feeding regimes and monitored their growth until maturation. Selection gradients calculated across studies show how maturation size is under net stabilizing selection in females and under directional selection in males. This pattern was used to predict that body size should be more canalized in females than in males. As expected, feeding affected male but not female maturation size. The sex-specific response of maturation size was related to a dramatic divergence between subadult male and female growth pathways. These results demonstrate the existence of sex-specific canalization and resource allocation to maturation size in this species, which causes variation in SSD depending on developmental conditions consistent with the differential-plasticity hypothesis explaining Rensch's Rule.     

The ecology, morphology and behaviour of Bakerdania elliptica (Acari: Prostigmata: Pygmephoridae), a mite associated with terrestrial isopods

In deciduous woodlands near to Bristol, South-west England, about one in ten specimens of the woodlouse Oniscus asellus (L.) are infested with Bakerdania elliptica (Krczal, 1959), a small pygmephorid mite of about 200 pm in length. Woodlice in the field rarely carry more than three mites, which are usually attached to the mid or lateral regions of the sixth and seventh pereonites on the ventral surface of the isopod. These 'favoured sites' correspond to regions of the exoskeleton of the woodlouse which are free from abrasion as they move through leaf litter. Mites removed from these areas and replaced on the first pereonite return to a favoured site, usually within 30 min. During this process they exhibit four distinct types of behaviour.]
Uninfested specimens of Oniscus usellus maintained in laboratory tanks on leaf litter from their 'own' site all become infested with mites within a week. The number of mites on the isopods increases rapidly under these conditions. After six weeks, each individual carries a mean of about 50 mites. The level of infestation is subsequently stable, probably due to saturation of favoured sites, since large numbers of unattached mites can be found in the leaf litter during this period.
Studies by light microscopy and scanning electron microscopy have shown that Bakerdania elliptica does not feed while attached to the cuticle of Oniscus asellus , and that all attached mites are females. Males are extremely rare and occur only in leaf litter. These observations suggest that the relationship between Bakerdania elliptica and Oniscus asellus is one of phoresy and that female mites use woodlice as an aid to dispersal during their life cycle.     

Warm developmental temperatures induce non-adaptive plasticity in the intrasexually selected colouration of a dragonfly

1. When the breeding environment fluctuates across generations, reproductive traits may evolve plasticity that optimises the balance between survival and mating success for the prevailing environment. 2. For sexually selected colouration, this balance can depend on environmental temperatures. Accordingly, breeding colouration often co-varies with temperature through space and time. However, whether such traits exhibit plasticity in response to environmental temperatures is poorly understood. 3. In the present study, a dragonfly (Pachydiplax longipennis) was reared under ambient or experimentally warmed conditions and tested for plasticity in its intrasexually selected wing colouration. Although wing colouration improves male territorial success, these advantages are smaller under warmer conditions than cooler conditions. It was therefore predicted that males reared under the ambient thermal conditions of the study site (Cleveland, Ohio) would develop more wing colouration than those reared under experimentally warmed conditions. 4. Contrary to this prediction, males reared in warm larval temperatures produced more wing colouration. Thus, although the secondary sexual colouration of this species displays some thermal plasticity, it does not appear to be adaptive relative to the known thermal variation of intrasexual selection in this population. 5. Given that the environment often determines the strength and direction of sexual selection, future studies should consider the potential for non-adaptive, and even maladaptive, developmental plasticity in the sexually selected traits of insects.     

Eco-morphological studies on pleopodal lungs and cuticle in Armadillidium species (Crustacea,Isopoda, Oniscidea)

Terrestrial isopods (Crustacea, Isopoda, Oniscidea) have adapted to land life by diverse morphological, physiological and behavioral changes. Woodlice species exhibit a large variety in this respect, their preferences ranging from moist to dry habitats. These moisture preference values are related to various morphological adaptations, rendering terrestrial isopods amenable to studying morphological adaptations to terrestrial life. We performed a comparison of four Armadillidium species (Armadillidium zenckeri, Armadillidium nasatum, Armadillidium versicolor, Armadillidium vulgare), by quantifying two morphological traits: the extent of the interfacial endothelium between the respiratory space and the hemolymph within pleopodal lungs and the thickness of tergite cuticle, which are ‘key factors’ in determining protection from desiccation. These values were measured from light micrographs of cross-sectioned lungs. The cosmopolitan A. vulgare, as a habitat generalist, seems to be the most resistant against desiccation and other environmental conditions, while A. zenckeri is the most sensitive one. Light microscopic studies revealed that the four species can be ordered similarly, if we compare them by the extension of the endothelial interface and cuticle thickness, suggesting that these morphological traits are important determinants of their distribution on habitat, microhabitat scales and through the existence of suitable habitats – together with many other factors – the geographical pattern of species occurence.     

Rearing Temperature Influences Adult Response to Changes in Mating Status

Rearing environment can have an impact on adult behavior, but it is less clear how rearing environment influences adult behavior plasticity. Here we explore the effect of rearing temperature on adult mating behavior plasticity in the butterfly Bicyclus anynana, a species that has evolved two seasonal forms in response to seasonal changes in temperature. These seasonal forms differ in both morphology and behavior. Females are the choosy sex in cohorts reared at warm temperatures (WS butterflies), and males are the choosy sex in cohorts reared at cooler temperatures (DS butterflies). Rearing temperature also influences mating benefits and costs. In DS butterflies, mated females live longer than virgin females, and mated males live shorter than virgin males. No such benefits or costs to mating are present in WS butterflies. Given that choosiness and mating costs are rearing temperature dependent in B. anynana, we hypothesized that temperature may also impact male and female incentives to remate in the event that benefits and costs of second matings are similar to those of first matings. We first examined whether lifespan was affected by number of matings. We found that two matings did not significantly increase lifespan for either WS or DS butterflies relative to single matings. However, both sexes of WS but not DS butterflies experienced decreased longevity when mated to a non-virgin relative to a virgin. We next observed pairs of WS and DS butterflies and documented changes in mating behavior in response to changes in the mating status of their partner. WS but not DS butterflies changed their mating behavior in response to the mating status of their partner. These results suggest that rearing temperature influences adult mating behavior plasticity in B. anynana. This developmentally controlled behavioral plasticity may be adaptive, as lifespan depends on the partner’s mating status in one seasonal form, but not in the other.     

Photoperiodic determination of gynoparae and males of damson-hop aphid Phorodon humuli

Abstract. Sexual morph production in Phorodon humuli is controlled by daylength. Wingless aphids reared from birth in short-day conditions (LD 12 : 12 h) and transferred when adult to long-day conditions (LD 18 : 6 h) produce only gynoparae and males until death some 7 weeks later, whereas those reared in long days and transferred to short days produce 20% wingless parthenogenetic females, 50% gynoparae and 30% of males in an overlapping sequence. No winged morph capable of re-infesting hop is produced. Less mature embryos are more sensitive to short days than older embryos because 100% of the former became gynoparae after 4 days of exposure of their mothers, and 59% when their mothers were exposed for the 4 days immediately preceding the birth of their offspring. Two days of exposure to short days switches 94% of young embryos from wingless to gynopara production when mature. The response to short days is irreversible. Wingless aphids reared from birth to adult in short days produce 30% fewer offspring than those reared to the same stage in long days and are male-biased, with 76–78% of their offspring being male.     

Limited flexibility in female Pacific field cricket (Teleogryllus oceanicus) exploratory behaviors in response to perceived social environment

How populations adapt, or not, to rapid evolution of sexual signals has important implications for population viability, but is difficult to assess due to the paucity of examples of sexual signals evolving in real time. In Hawaiian populations of the Pacific field cricket (Teleogryllus oceanicus), selection from a deadly parasitoid fly has driven the rapid loss of a male acoustic signal, calling song, that females use to locate and evaluate potential mates. In this newly quiet environment where many males are obligately silent, how do phonotactic females find mates? Previous work has shown that the acoustic rearing environment (presence or absence of male calling song) during late juvenile stages and early adulthood exposes adaptive flexibility in locomotor behaviors of males, as well as mating behaviors in both sexes that helps facilitate the spread of silent (flatwing) males. Here, we tested whether females also show acoustically induced plasticity in walking behaviors using laboratory‐reared populations of T. oceanicus from Kauai (HI; >90% flatwings), Oahu (HI; ~50% flatwings), and Mangaia (Cook Islands; no flatwings or parasitoid fly). Though we predicted that females reared without song exposure would increase walking behaviors to facilitate mate localization when song is rare, we discovered that, unlike males, female T. oceanicus showed relatively little plasticity in exploratory behaviors in response to an acoustic rearing environment. Across all three populations, exposure to male calling song during development did not affect latency to begin walking, distance walked, or general activity of female crickets. However, females reared in the absence of song walked slower and showed a marginally non‐significant tendency to walk for longer durations of time in a novel environment than those reared in the presence of song. Overall, plasticity in female walking behaviors appears unlikely to have facilitated sexual signal loss in this species.     

Using woodlice (Isopoda,Oniscoidea) to demonstrate orientation behaviour

Woodlice show two behavioural kineses as adaptations to prevent water loss. These are attraction to solid objects, including each other, (thigmokinesis), and attraction to dark places (negative photqkinesis). The interaction between these two kineses was tested in a classroom experiment, in which one to five woodlice were placed in a petri dish with one half painted black. The strongest response was the thigmotactic attraction, with negative photokinesis playing a secondary role. From these experiments students learn about cooperation, taxes (directional responses to stimulus i.e. the woodlice move towards or away from a stimulus) and kineses (non-directional response to stimulus, i.e. the woodlice respond by showing an increase or decrease in activity level), and experimental design.     

Aggressive Transition between Alternative Male Social Tactics in a Long-Lived Australian Dragon (Physignathus lesueurii) Living at High Density

Theory predicts the evolution of alternative male social tactics when intense competition coupled with the superior competitive ability of some individuals limits access to reproductive opportunities by others. How selection has shaped alternative social tactics may be especially interesting in long-lived species where size among sexually mature males varies markedly. We conducted experimental studies on long-lived eastern Australian water dragons living where competition was intense to test the hypotheses that mature males adopt alternative social tactics that are plastic, and that large size and body condition determine resource-holding potential. Approximately one-half of mature males (N = 14) defended territories using high rates of patrol and advertisement display, whereas 16 smaller mature males having lower body condition indices utilized non-territorial social tactics. Although territorial males were larger in absolute size and head dimensions, their heads were not allometrically larger. Territorial males advertised very frequently using displays involving stereotypical movements of the head and dewlap. More aggressive displays were given infrequently during baseline social conditions, but increased during periods of social instability. Female home ranges overlapped those of several territorial and non-territorial males, but females interacted more frequently with territorial males. The extreme plasticity of social tactics in this species that are dependent on body size was confirmed by two instances when relatively large non-territorial males spontaneously evicted territory owners, and by marked shifts in tactics by non-territorial males in response to temporary experimental removals of territory owners, followed (usually) by their expulsion when original owners were reinstated. The high level of social plasticity in this population where same-sex competitors are densely concentrated in preferred habitat suggests that chronic high energetic costs of defense may select for males to cycle between territorial and non-territorial social tactics depending upon their changing energetic status and their current capacity for competition with rivals.     

Variable male potential rate of reproduction: high male mating capacity as an adaptation to parasite-induced excess of females?

Numerous animals are known to harbour intracytoplasmic symbionts that gain transmission to a new host generation via female eggs and not male sperm. Bacteria of the genus Wolbachia are a typical example. They infect a large range of arthropod species and manipulate host reproduction in several ways. In terrestrial isopods (woodlice), Wolbachia are responsible for converting males into females (feminization (F)) in some species, or for infertility in certain host crosses in other species (cytoplasmic incompatibility (CI)). Wolbachia with the F phenotype impose a strong excess of females on their host populations, while Wolbachia expressing CI do not. Here, we test the possibility that male mating capacity (MC) is correlated with Wolbachia-induced phenotype. We show that males of isopod hosts harbouring F Wolbachia possess a strong MC (i.e. are able to mate with several females in a short time), while those of species harbouring CI Wolbachia possess a weaker MC. This pattern may be explained either by the selection of high MC following the increase in female-biased sex ratios, or because the F phenotype would lead to population extinction in species where MC is not sufficiently high. This last hypotheses is nevertheless more constrained by population structure.     

Developmental temperature affects the expression of ejaculatory traits and the outcome of sperm competition in Callosobruchus maculatus

The outcome of post‐copulatory sexual selection is determined by a complex set of interactions between the primary reproductive traits of two or more males and their interactions with the reproductive traits of the female. Recently, a number of studies have shown the primary reproductive traits of both males and females express phenotypic plasticity in response to the thermal environment experienced during ontogeny. However, how plasticity in these traits affects the dynamics of sperm competition remains largely unknown. Here, we demonstrate plasticity in testes size, sperm size and sperm number in response to developmental temperature in the bruchid beetle Callosobruchus maculatus. Males reared at the highest temperature eclosed at the smallest body size and had the smallest absolute and relative testes size. Males reared at both the high‐ and low‐temperature extremes produced both fewer and smaller sperm than males reared at intermediate temperatures. In the absence of sperm competition, developmental temperature had no effect on male fertility. However, under conditions of sperm competition, males reared at either temperature extreme were less competitive in terms of sperm offence (P₂), whereas those reared at the lowest temperature were less competitive in terms of sperm defence (P₁). This suggests the developmental pathways that regulate the phenotypic expression of these ejaculatory traits are subject to both natural and sexual selection: natural selection in the pre‐ejaculatory environment and sexual selection in the post‐ejaculatory environment. In nature, thermal heterogeneity during development is commonplace. Therefore, we suggest the interplay between ecology and development represents an important, yet hitherto underestimated component of male fitness via post‐copulatory sexual selection.     

Influence of male nuptial plumage on mate selection in the female mallard (Anas platyrhynchos)

The aim of this study was to investigate the basis of mate selection in the mallard, with special reference to the role of the female. Females were reared under more or less restricted social conditions during ontogeny. When sexual mature they were offered a choice between males of different colour categories. The females used were raised under three conditions: (1) with natural mother and siblings (control females), (2) with experience of other conspecific females but no males, or (3) with no experience of ducks at all (Kaspar Hauser). The total number of females used was 227. The females were able to discriminate between normal mallards and differently coloured mallards to a large extent irrespective of their own early experience. This suggests that the females' initial mate selection is dependent on male plumage characteristics.     

Effects of <Emphasis Type="Italic">Acanthocephalus</Emphasis> sp. (Acanthocephala: Echinorhynchidae) on the body size and reproduction of isopods (<Emphasis Type="Italic">Asellus hilgendorfi</Emphasis>)

 Isopods Asellus hilgendorfi were collected from a small lake in northern Japan and examined to determine whether their body size and reproduction were affected by infection with larval acanthocephalans (Acanthocephalus sp.). Seasonal changes in the breeding ratio of isopods and the prevalence of larval acanthocephalan infection showed a reverse trend. Acanthocephalan larvae occurred mainly in males and immature females and were rarely found in mature females. In late immature females, the body size, as indicated by the width of the pleotelson, of infected isopods was significantly larger than that of uninfected ones. These results suggest that acanthocephalans can prevent female isopods from attaining sexual maturity and increasing their body size. Received: January 9, 2002 / Accepted: December 16, 2002 Acknowledgments We thank Professor Shōichi Saito, Faculty of Education, Hirosaki University, for his encouragement of the present study. Thanks are also due to the Iwasaki Village Office and the Fukaura Forestry Office for giving us permission for the survey. Correspondence to:A. Ohtaka