<Emphasis Type="Italic">Uncisudis posteropelvis</Emphasis>, a new species of barracudina (Aulopiformes: Paralepididae) from the western North Pacific Ocean

Six specimens (2 flexion larvae: 9.5–10.4mm in notochord length; 4 postflexion larvae: 12.3–18.2mm in standard length) collected from the western North Pacific are tentatively ascribed to the genus Uncisudis of the tribe Lestidiini of the subfamily Paralepidinae (Paralepididae) in sharing remarkably elongate and filamentous pelvic fin rays, their tips reaching the origin of the anal fin. They are described as Uncisudis posteropelvis sp. nov. in uniquely having the insertion of pelvic fins closer to the origin of anal fin than to the posterior end of dorsal fin base among lestidiine species. Addition to this character, the new species has remarkably elongate and filamentous dorsal fin rays, the short distance between anus and origin of anal fin (4.2–6.1% of standard length, SL), the posteriorly located pelvic fins (prepelvic length 69.4–71.5% SL), dorsal fin rays 10, anal fin rays 28–29, myomeres 41–42+38–40=80–81 (vertebrae 38+41=79), and peritoneal pigment spots 11–12. The occurrence of larvae differing in pigment pattern from the present new species suggests another undescribed species of Uncisudis in the western South Pacific.     

Effects of constant and fluctuating temperatures on sporulation and infection by the aphid-pathogenic fungus Pandora neoaphidis

Laboratory studies were performed to assess the importance of temperature on sporulation and infection by the aphid-pathogenic fungus Pandora neoaphidis (Remaudière and Hennebert) Humber. Numbers of primary conidia discharged from mycelium formulated as alginate granules and unformulated mycelial mats were assessed, as well as infection of the potato aphid, Macrosiphum euphorbiae (Thomas) (Homoptera, Hemiptera, Aphididae), using culture plugs as inoculum sources. Sporulation from experiments at constant temperatures indicated the optimum temperature range was 10–20°C for both mycelial preparations and there was no or very little sporulation at 30°C. Infection of aphids kept at 15°C was 34–50%, while infection at 25°C was 11–44%. At 20°C, 77–79% of aphids were infected. Under fluctuating temperature cycles, conidia numbers did not differ when mycelial preparations were maintained at 18–25°C compared with 18–20°C, but fewer conidia were recorded when preparations were exposed continuously to 18–30°C. Infections of inoculated aphids kept for varying numbers of days at 18–25°C varied between 24–47%, but only 3–32% of aphids were infected when exposed to a cycle of 18–30°C for various times. Unformulated mycelial mats of P. neoaphidis appear to be superior to forumlated alginate granules for use in experimental greenhouse and field trials, since temperature stability is similar for both materials but mycelial mats are much easier to produce.     

The Effects of Hypoxia on Three Sympatric Shark Species: Physiological and Behavioral Responses

Behavioral and physiological responses to hypoxia were examined in three sympatric species of sharks: bonnethead shark Sphyrna tiburo, blacknose shark, Carcharhinus acronotus, and Florida smoothhound shark, Mustelus norrisi, using closed system respirometry. Sharks were exposed to normoxic and three levels of hypoxic conditions. Under normoxic conditions (5.5–6.4mg l^–1), shark routine swimming speed averaged 25.5 and 31.0cm s^–1 for obligate ram-ventilating S. tiburo and C. acronotus respectively, and 25.0cm s^–1 for buccal-ventilating M. norrisi. Routine oxygen consumption averaged about 234.6 mg O₂kg^–1h^–1 for S. tiburo, 437.2mg O₂kg^–1h^–1 for C. acronotus, and 161.4mg O₂ kg^–1 h^–1 for M. norrisi. For ram-ventilating sharks, mouth gape averaged 1.0cm whereas M. norrisi gillbeats averaged 56.0 beats min^–1. Swimming speeds, mouth gape, and oxygen consumption rate of S. tiburo and C. acronotus increased to a maximum of 37–39cm s^–1, 2.5–3.0cm and 496 and 599mg O₂ kg^–1 h^–1 under hypoxic conditions (2.5–3.4mg l^–1), respectively. M. norrisi decreased swimming speeds to 16cm s^–1 and oxygen consumption rate remained similar. Results support the hypothesis that obligate ram-ventilating sharks respond to hypoxia by increasing swimming speed and mouth gape while buccal-ventilating smoothhound sharks reduce activity.     

Inoculation of sugar mill by-products compost with N2-fixing bacteria

Inoculation of sugar mill by-products compost with N₂-fixing bacteria may improve its quality by increasing total N and available P. Compost was inoculated with Azotobacter vinelandii(ATCC 478), Beijerinckia derxii (ATCC 49361), and Azospirillumsp. TS8, each alone and all three together. Numbers of all N₂-fixing bacteria in compost declined from an initial population of 5×10⁵cellsg^–1 during incubation. The population of Azotobacter declined to approximately 2×10²cellsg^–1 and the population of Beijerinckia and Azospirillum declined to approximately 9×10³ and 3.5×10⁴cellsg^–1 respectively, at day 50. Inoculation with N₂-fixing bacteria increased acetylene reduction, total N by 6–16 and available P by 25–30% in comparison to the uninoculated control. Increasing the N content and P availability of compost increases its value and there may be additional benefit from providing N₂ fixing bacteria.     

Carbon dynamics and budget in a <Emphasis Type="Italic">Miscanthus sinensis</Emphasis> grassland in Japan

We investigated the carbon dynamics and budget in a grassland of Miscanthus sinensis, which is widely distributed in Japan, over a 2-year period (2000–2001). Plant biomass began to increase from May and peaked in September, then decreased towards the end of the growing season (October). Soil respiration rates also exhibited seasonal fluctuations that reflected seasonal changes in soil temperature and root respiration. The contribution of root respiration to total soil respiration was 22–41% in spring and summer, but increased to 52–53% in September. To determine the net ecosystem production (carbon budget), we estimated annual net primary production, soil respiration, and root respiration. Net primary production was 1207 and 1140gCm^–2 in 2000 and 2001, respectively. Annual soil respiration was 1387gCm^–2 in 2000 and 1408gCm^–2 in 2001; root respiration was 649 and 695gCm^–2 in 2000 and 2001, respectively. Moreover, some of the carbon fixed as net production (457–459gCm^–2) is removed by mowing in autumn in this grassland. Therefore, the annual carbon budget was estimated to be –56gCm^–2 in 2000 and – 100gCm^–2 in 2001. These results suggest that the Miscanthus sinensis grassland in Japan can act as a source of CO₂.     

Uptake of sewage derived nitrogen by<Emphasis Type="Italic"> Ulva rigida</Emphasis> (Chlorophyceae) in Bahía Nueva (Golfo Nuevo,Patagonia, Argentine)

Uptake kinetics of nitrogen derived from sewage–seawater mixtures (2.5–20% v/v effluent) were determined in the laboratory for Ulva rigida (Chlorophyceae) native from Bahía Nueva (Golfo Nuevo, Patagonia, Argentine). In terms of nitrogen concentration, experimental enrichment levels varied between 53.7 and 362.3M of ammonium and between 0.77 and 6.21M of nitrate+nitrite. Uptake rates were fitted to the Michaelis–Menten equation, with the following kinetic parameters: ammonium: V_max = 591.2molg^–1h^–1, K _s=262.3M, nitrate+nitrite: V _max=12.9molg^–1h^–1, K _s=3.5M). Both nutrients were taken up simultaneously, but ammonium incorporation was faster in all cases. The results show a high capability of Ulva rigida to remove sewage-derived nitrogen from culture media. In the field, most of the nitrogen provided by the effluent would be tied up in algal biomass, supporting low nitrogen levels found at a short distance away from the source.     

In vitro propagation of Eucomis L'Hérit species - plants with medicinal and horticultural potential

Eucomis (Family Hyacinthaceae) are deciduousgeophytes with long, narrow leaves and erect, densely packed flower spikes. Thebulbs are greatly valued in traditional medicine for the treatment of a variety ofailments, and are thus heavily harvested for trade in South Africa's traditional herbal markets. Eucomis species propagaterelatively slowly from offsets and seed, and this, together with theirover-utilisation ethnopharmacologically, has led to their threatened status. Thein vitro propagation of the genusEucomis was investigated to optimise this technique for thebulk production of plants for commercial and conservation purposes. Multipleshoot production was initiated from leaf explants, in all species studied. AMurashige and Skoog (MS) medium, supplemented with 100mgamp;ell;^–1 myo-inositol, 20gamp;ell;^–1 sucrose, and solidified with 2gamp;ell;^–1 Gelrite® was used. Theoptimal hormone combination for shoot initiation in the majority of species was1 mgamp;ell;^–1 NAA and 1mgamp;ell;^–1 BA. Optimal root initiation wasdemonstrated on media supplemented with 1mgamp;ell;^–1 IAA, IBA or NAA, depending onspecies. A continuous culture system using this protocol produced 25–30plantlets per culture bottle, with 10–25 specimens per bottle availablefor acclimatisation. To maximise plantlet survival, different support media usedduring the acclimatisation process were necessary. Certain species respondedbest on a vermiculite medium, while perlite (which holds less water) wasnecessary for the optimal survival rate of other species. Acclimatised plantletswere repotted in a sand: soil mix (1:1).     

Increased erythritol production in Torula sp. by Mn2+and Cu2+

The production of erythritol and the erythritol yield from glucose by Torula sp. were improved, in increasing order, by supplementing with 10 mg MnSO₄4H₂O l^–1, 2 mg CuSO₄5H₂O l^–1, and both 10 mg MnSO₄4H₂O l^–1 and 2 mg CuSO₄5H₂O l^–1. Mn²⁺ decreased the intracellular concentration of erythritol, whereas Cu²⁺ increased the activity of erythrose reductase in cells. These results suggest that Mn²⁺ altered the permeability of cells, whereas Cu²⁺ increased the activity of erythrose reductase in cells.     

Indirect method to estimate convective gas flow through culms of a <Emphasis Type="Italic">Phragmites australis</Emphasis> stand

We propose an indirect method to estimate continuously the rate of convective gas flow in a Phragmites australis stand. In this method, the rate of gas flow is estimated using the dynamic pressure differential in a culm and the convective conductance of the culm. The rate of gas flow obtained by this indirect method coincided well with that obtained by the direct method in which a culm is detached and then reconnected to the stubble using a mass flow meter. We monitored the total gas flux through a P. australis stand in a field and found that it fluctuated diurnally with the dynamic pressure differential in culms, showing a highest rate of 26lairm^–2 ground area h^–1 at noon. The total daily gas flux was about 170lairm^–2. Our indirect method has advantages in simultaneous and continuous measurements for a cluster of culms. This method will be of use not only to quantify various gas dynamics through aquatic plants in aquatic ecosystems but also to elucidate the ecosystem processes and properties that regulate these gas dynamics.     

Developmental morphology of the cyprinid fish Chela dadiburjori

Embryonic, larval, and juvenile development of an Indian cyprinid fish, Chela dadiburjori, is described from laboratory-reared specimens. The eggs, measuring 0.7–0.9mm in diameter, were demersal, almost spherical in shape, transparent and unpigmented, with a pale yellow yolk and no oil globule. Hatching occurred 50–61h after fertilization at ca. 27°C. The newly hatched larvae, measuring 2.4–2.6mm in body length (BL), had melanophores on the body with 14–16+14–17=29–31 myomeres. Two dark transverse bands on the ventral body surface and one melanophore on the lower margin of the eye in newly hatched larvae were diagnostic. Additionally, a cement organ for adhering to objects was present on the forehead of yolk sac larvae <3.1mm BL. The yolk was completely absorbed at 3.5mm BL. Notochord flexion was initiated at 5.0mm BL and finished at 6.0mm BL. Aggregate numbers of all fin rays were completed at 9.2mm BL. Squamation was initiated on the caudal peduncle at 8.0mm BL and completed at 10mm BL. The eggs of C. dadiburjori resembled those of the closely related species Devario malabaricus and Danio rerio. The larvae and juveniles of C. dadiburjori were also similar to those of the latter species in general morphology, especially the presence of body melanophores in newly hatched individuals and a distinctive lateral streak on the head during the period from yolk sac to postflexion larvae. However, early yolk sac larvae of C. dadiburjori were more similar to those of Devario malabaricus than Danio rerio in having a cement organ on the forehead. Larvae and juveniles of C. dadiburjori differed from those of the latter two species in pigmentation on the ventral body surface at hatching and around the mouth during the period from preflexion to early postflexion larvae and in having a dark lateral streak or band on the body in postflexion larvae and juveniles.     

Structure and dynamics of a <Emphasis Type="Italic">Castanopsis cuspidata</Emphasis> var. <Emphasis Type="Italic">sieboldii</Emphasis> population in an old-growth,evergreen, broad-leaved forest: The importance of sprout regeneration

The population structure and dynamics of Castanopsis cuspidata var. sieboldii were studied to evaluate vegetative and sapling regeneration in an old-growth, evergreen broad-leaved forest exposed to low-severity typhoon disturbances by annual typhoons on the Tsushima Islands in Japan. The density of individuals 5cm d.b.h. was 38.0ha^–1 in 1990; 7.9% were multiple-stemmed individuals. Over the 7-year study period (1990–1997), the number of individuals decreased, although the number of stems increased. Over 30% of apparently dead individuals were reconstituted by sprouting stems. Compared with sprout regeneration, sapling regeneration rarely occurred and was only observed in canopy gaps. Most individuals had at least one sprout shoot (H 30cm, d.b.h. <5cm), and the number and size of sprout shoots increased as the size of the individuals increased. During the study period, larger individuals with stem breakage tended to produce sprout stems. The density of saplings was 1074ha^–1 and they were more abundant in canopy gaps than under closed canopies, but large saplings were very rare even in canopy gaps. The population of C.cuspidata var. sieboldii consisted primarily of single-stemmed individuals with a few multiple-stemmed individuals providing a sprout bank. Larger individuals responded to the low severity typhoon disturbances and formed sprout stems. Although many saplings were observed, regeneration occurred more often by sprout formation than by growth of saplings. Thus, sprout regeneration is an important mode of regeneration, which allows this pioneer-like species to maintain its population in this forest.     

The Effects of Increasing UV-B Radiation on Pigmentation, Growth and Survival of Coregonid Embryos and Larvae

In the northern regions UV-B radiation levels have increased due to ozone depletion. A two-week laboratory experiment was conducted to measure the effects of UV-B radiation on the pigmentation, growth, oxygen consumption rate and survival of whitefish, Coregonus lavaretus, and vendace, Coregonus albula, larvae. In May newly hatched embryos were exposed in laboratory aquaria to three CIE weighted UV-B radiation levels: subambient (daily dose 1.37kJm^–2), 9% (1.81kJm^–2) and 34% higher (2.24kJm^–2) than ambient. Control embryos and larvae were not exposed to UV-B. Larvae of whitefish and vendace that were irradiated with highest UV-B level had 32% and 31% more melanin than control larvae, respectively, which we interpret as an apparent induced response. In controls, the species difference revealed 53% more melanin in vendace larvae than in whitefish larvae. UV-B radiation had no effect on the mortality of either species, the survival being high in all treatments (>90%). Additionally, neither growth rate nor the metabolic rate of larvae of either species was affected by UV-B radiation. Thus, in relation to future scenarios UV-B radiation may not be a threat to whitefish or vendace larvae in current or expected radiation levels.     

Control of basidiospore production by a nuclear gene in the fungus Coprinus congregatus

Summary The genetical control of basidiospore production by sporophores of the fungus Coprinus congregatus was studied. This species is characterized by a bipolar compatibility control, and homokaryons with complementary alleles A1 and A2 can be distinguished apart. We confirmed that the pale mushroom phenotype of the fungus is determined by a nuclear gene symbolized pal. This gene also controls a sporeless character and segregates independently of the mating-type locus. Dikaryons homoallelic for the pal ^– allele produce typical pale and sporeless sporophores, while heteroallelic (pal ⁺, pal ^–) and homoallelic (pal ⁺, pal ⁺) dikaryons produce normal or almost normal sporulating sporophores. In order to segregate homokaryons homoallelic for the pal gene (A1, pal ^–; A1, pal ⁺, A2, pal ^–; A2, pal ⁺), the following protocols were used: (a) the dikaryotization of stock homokaryons containing the pal ⁺ allele and of each mating type, A1 or A2, by dikaryotic mycelia homoallelic for the pal ^– allele; (b) the culturing of homokaryotic mycelia issuing from the germination of basidiospores from sporophores produced by dikaryotic mycelia heterokaryotic for the pal gene; (c) the culturing of mycelia grown from protoplasts obtained from dikaryons homoallelic for the pal ^– allele (D6 strain), and from homokaryons heteroallelic for the pal gene (H8), or homoallelic for pal ^#x002B;+ allele (H7). These techniques enabled us to segregate homokaryons of the four types defined above and were indispensable in the segregation of the pal ^– homoallelic homokaryons as no basidiospores were produced by typical pale mushrooms.     

Proliferation and germination of somatic embryos from embryogenic suspension cultures in <Emphasis Type="Italic">Phœnix dactylifera</Emphasis>

The present study demonstrates a procedure for the rapid development of a high number of somatic embryos from embryogenic suspension culture. This method might be efficient for mass propagation of Phnix dactylifera L. Embryogenic callus placed in liquid medium with 10^–5M ABA yielded an average 72 embryos per 100ml of culture medium within 2months, while those placed on solid medium yielded an average of 33, 20 and 16 embryos per 100ml of culture medium respectively for 10^–7, 10^–6 and 10^–5 M ABA after 4months. The combination of 2,4-DIchlorophenoxyacetic acid (2,4-D) (4.5×10^–7M), glutamine (6.7×10^–4M), and ABA (10^–5M) (L8 liquid medium) showed a beneficial effect on somatic embryos production compared to 2,4-D and glutamine alone, while this combination significantly (p<0.05) increased the accumulation of storage proteins (144 and 138mgg^–1 DW respectively for Jihel and Bousthami noir cultivars) in somatic embryos. The somatic embryos which underwent maturation on medium containing only 4.5×10^–7M 2,4-D and 10^–5M ABA (L6 liquid medium) accumulated more sugars (292 and 265mgg^–1 DW respectively for Jihel and Bousthami noir) than those matured on any other liquid medium. Histological studies revealed that somatic embryos (developed in L6 and L8 liquid media) accumulated less reserve compounds (proteins and sugars) than zygotic embryos. The addition of activated charcoal (0.25 and 0.5gl^–1) and phytagel^® (2.5gl^–1) to the germination medium may be useful for enhancing the germination of Phnix dactyliferasomatic embryos.     

Population density and group composition of Japanese sika deer (<Emphasis Type="Italic">Cervus nippon yakushimae</Emphasis>) in an evergreen broad-leaved forest in Yakushima,southern Japan

The population density of Japanese sika deer (Cervus nippon yakushimae Kuroda and Okada) in an evergreen broad-leaved forest in Yakushima, southern Japan, was surveyed over 4years from 1998 to 2001. Two approximately 50ha study sites, Hanyama and Kawahara, were established with a total of 4km of census trails at each site. The estimated densities of sika deer at the two sites were 43–70 deerkm^–2 at Hanyama and 63–78 deerkm^–2 at Kawahara, although these values might be underestimates. The adult sex ratio (number of adult males:number of adult females) ranged from 0.6 to 1.0 at Hanyama, and from 0.4 to 0.9 at Kawahara. Mean group size was 1.9 deer (male group, 1.5 deer; female group, 1.6 deer; mixed group, 3.6 deer). The population density of sika deer was relatively high compared to other sites in Japan, with the exception of very small (<10km²) islands. Possible explanations for this naturally high density of sika deer in an evergreen broad-leaved forest in Yakushima are discussed.     

Accumulation and depuration of microcystin produced by the cyanobacterium <Emphasis Type="Italic">Microcystis</Emphasis> in a freshwater snail

Seasonal changes in microcystin concentrations in a resident snail (Sinotaia histrica) and an edible clam (Corbicula sandai) in Lake Biwa were surveyed. To clarify both the accumulation and depuration of microcystins, experimental studies with microcystin were also carried out on the snail. In the field investigation, microcystin was detected from the hepatopancreas and intestine of S. histrica (up to 3.2µgg^–1 dry weight and 19.5µgg^–1 dry weight, respectively); however, no microcystin was detected in the hepatopancreas of C. sandai. In the laboratory experiment, the microcystin-LR concentration in the hepatopancreas of S. histrica reached a value of 436µgg^–1 dry weight on day 10 of 15 days of uptake, and a high value persisted despite a depuration period of 15 days. The depuration rate constant of microcystin and its biological half-life were 0.0828 day^–1 and 8.4 days, respectively. These results indicate that S. histrica has a high ability to accumulate microcystin in its tissue. Because S. histrica is predated by fish and water fowl, it is likely to play an important role as a vector for microcystin in lakes with dense blooms of toxic cyanobacteria.     

Laboratory culture studies of Trichodesmium isolated from the Great Barrier Reef Lagoon, Australia

Cultures of Trichodesmium from the Northern and Southern Great Barrier Reef Lagoon (GBRL) have been established in enriched seawater and artificial seawater media. Some cultures have been maintained with active growth for over 6years. Actively growing cultures in an artificial seawater medium containing organic phosphorus (glycerophosphate) as the principal source of phosphorus have also been established. Key factors that contributed to the successful establishment of cultures were firstly, the seed samples were collected from depth, secondly, samples were thoroughly washed and thirdly, incubations were conducted under relatively low light intensities (PAR 40–50molquantam^–2s^–1). N₂ fixation rates of the cultured Trichodesmium were found to be similar to those measured in the GBRL. Specific growth rates of the cultures during the exponential growth phase in all enriched media were in the range 0.2–0.3day^–1 and growth during this phase was characterised by individual trichomes (filaments) or small aggregations of two to three trichomes. Characteristic bundle formation tended to occur following the exponential growth phase, which suggests that the bundle formation was induced by a lack of a necessary nutrient e.g. Fe. Results from some exploratory studies showed that filament-dominated cultures of Trichodesmium grew over a range of relatively low irradiances (PAR 5–120molquantam^–2s^–1) with the maximum growth occurring at 40–50molquantam^–2s^–1. These results suggest that filaments of the tested strain are well adapted for growth at depth in marine waters. Other studies showed that growth yields were dependent on salinity, with maximum growth occurring between 30 and 37psu. Also the cell yields decreased by an order of magnitude with the reduction of Fe additions from 450 to 45nM. No active growth was observed with the 4.5nM Fe addition.     

Mode of uptake of sterol by Arthrobacter simplex

The mechanism of uptake of water-insoluble -sitosterol by a newly isolated strain of Arthrobacter simplex SS-7 was studied. The production of an extracellular sterol-pseudosolubilizing protein during growth of A. simplex on -sitosterol was demonstrated by isolating the factor from the cell-free supernatant and its subsequent purification by Sephadex G-150 column chromatography. The M _r of the purified sterol-pseudosolubilizing protein determined by SDS–PAGE was 19kDa. The rate of sterol pseudosolubilization (5.2×10^–3g l^–1h^–1) could not adequately account for the rate of sterol uptake (72×10^–3g l^–1h^–1) and the specific growth rate (56×10^–3 h^–1). However in the unfavourable growth condition, when the cells were treated with sodium azide at the level of 30–60% of MIC, the sterol pseudosolubilization accounted for nearly 74% of the total growth containing 96% free cells. Cellular adherence to substrate particles was found to play an active role in the normal growth of the strain on -sitosterol. Unlike sodium acetate-grown cells, whose surface activity was negligible (60mNm^–1), the sterol-grown cells had strong surface activity (40mNm^–1). The high lipid content and long chain fatty acids in the cell-wall of -sitosterol-grown cells probably contribute to the high sterol adherence activity of the cells.     

Functional expression of keratinase (kerA) gene from Bacillus licheniformis in Pichia pastoris

A 1.2 kb DNA fragment coding for the pro-peptide and mature keratinase from Bacillus licheniformis PWD-1 (kerA) was cloned into vectors pPICZA and pGAPZA for extracellular expression in the methylotrophic yeast, Pichia pastoris. Recombinant keratinase was secreted by the pPICZA-kerA transformants 24 h after methanol induction of shake-flask cultures, and reached a final yield of 124 mg l^–1 (285 U ml^–1) 144 h after the induction. The recombinant keratinase was glycosylated ( 39 kDa), and was optimal between pH 8.5–9.5 and between 55°C –60°C using azokeratin as substrate. The enzyme degraded bovine serum albumin, collagen, and soy protein concentrate. In conclusion, P. pastoris can be used as an efficient host to express keratinase for nutritional and environmental applications.     

Effects of outbreak of the train millipede Parafontaria laminata armigera Verhoeff (Diplopoda: Xystodesmidae) on litter decomposition in a natural beech forest in Central Japan. 1. Density and biomass of soil invertebrates

Soil invertebrates were studied to determine the animals important in litter decomposition and soil formation in a natural forest of Fagus crenata Blume mixed with Quercus mongolica var. grosseserrata Rehd. et Wils., where outbreak of the adult train millipede, Parafontaria laminata armigera Verhoeff was recorded in 1980. The density and biomass of soil invertebrates, and their distribution in soil, were surveyed largely by hand sorting and also using Tullgren and Baermann apparatuses. The densities of soil macroinvertebrates were 10²–10³ individuals/m². Diplopoda was the most abundant among them, 87.3% of which was the train millipede. The total biomass in wet weight of soil macro-invertebrates was 34.9–70.5g/m² in 1980–1981, when the millipede was pre-adult or adult, and it decreased to 3.5–6.7g/m² in 1982–1984, when it was young. The density of soil invertebrates extracted by apparatuses was 10⁴–10⁶ individuals/m². The total biomass of these was estimated to be 5.8–10.8g/m². The majority of soil invertebrates lived in the A₀ horizon, whereas the train millipede lived in soil of the A₁+A₂ horizon. These results suggest that the train millipede is one of the most important soil invertebrates affecting soil properties. For evaluating the importance of the species, more than 8 years survey of hand-sorting together with the Tullgren apparatus was necessary, as the train millipede grew synchronously and had an 8-year life span and young stages of 1.5–3.5mm long.