Fifth International Nematology Symposium. Vladivostok 2003

Fifth International Nematology Symposium. Vladivostok 2003. http://wwwimb.dvo.ru/misc/fisn/index.htm

RUSSIAN SOCIETY OF NEMATOLOGISTS

FIFTH INTERNATIONAL NEMATOLOGY SYMPOSIUM

VLADIVOSTOK, RUSSIA, 13 - 17 JULY 2003

| ABSTRACTS | ORGANIZING COMMITTEE |

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RUSSIAN SOCIETY OF NEMATOLOGISTS

FIFTH INTERNATIONAL NEMATOLOGY SYMPOSIUM

VLADIVOSTOK, RUSSIA, 13 - 17 JULY 2003

Organizing committee of the Fifth English Language International Symposium of the Russian Society of Nematologists in Vladivostok, Russia 2003

Dr. Vladimir V.Yushin Chairman of Organising Committee President of the Russian Society of Nematologists Institute of Marine Biology FEB RAS Vladivostok, RUSSIA E-mail: [email protected]

Dr. Sergei E. Spiridonov Secretary and treasurer of the Russian Society of Nematologists Institute of Parasitology RAS Moscow, RUSSIA E-mail: [email protected]

Dr. Sergei A. Subbotin IFNS Rep. - Russian Society of Nematologists Institute of Parasitology RAS Moscow RUSSIA E-mail: [email protected]

Dr. Prof. Vladimir L. Kasyanov Academician of RAS Director of the Institute of Marine Biology RAS Director of the Academy of Ecology, Marine Biology and Biotechnology FESU Director of the Scientific Division of the Science and Education Program "Marine Biota" FESU Vladivostok, RUSSIA E-mail: [email protected]

Dr. Alex Yu. Ryss Zoological Institute RAS Saint-Petersburg, RUSSIA E-mail: [email protected]

Dr. Prof. Vladimir V. Malakhov Corresponding member of RAS Moscow State University Moscow, RUSSIA E-mail: [email protected]

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Dr. Prof. Alexei V. Tchesunov Moscow State University Moscow, RUSSIA E-mail: [email protected]

Dr. Prof. Derek J.F. Brown Elected Fellow of the Russian Society of Nematologists Mycology, Bacteriology & Nematology Unit Scottish Crop Research Institute Invergowrie, Dundee, Scotland, U.K.

Dr. Liao Jinling IFNS Rep. - Chinese Society of Plant Nematologists Laboratory of Plant Nematology South China Agricultural University Guangzhou 510642, CHINA E-mail: [email protected] or [email protected]

Dr. Mutsuhiro Yoshida The Japanese Nematological Society Nematology & Soil Zoology Unit Department of Biological Safety National Institute of Agro-Environmental Sciences Kannonndai, Tsukuba, Ibaraki, JAPAN E-mail: [email protected]

FINS Local Organazing Committee in Vladivostok

Institute of Marine Biology FEB RAS

Dr. Vladimir V.Yushin, Chairman of Organising Committee E-mail: [email protected] E-mail: Dr. Lyudmila S.Belogurova [email protected] Dr. Olga N. Pavlyuk E-mail: [email protected]

Institute of Biology and Soil Sciences FEB RAS E-mail: [email protected]

Dr. Alexander S. Eroshenko Dr. Irina A. Kruglik Dr. Inna P. Kazachenko Dr. Tamara V. Volkova

Academy of Ecology, Marine Biology and Biotechnology of the Far Eastern State University

Dr. Olga I. Dashchenko E-mail: [email protected] Inna L. Davydkova E-mail: [email protected] Nataly L. Demchenko E-mail: [email protected] Dr. Nataly P. Fadeeva E-mail: [email protected] Vladimir V. Morduchovich E-mail: [email protected] Yulia K. Zograf E-mail: [email protected]

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RUSSIAN SOCIETY OF NEMATOLOGISTS

FIFTH INTERNATIONAL NEMATOLOGY SYMPOSIUM

VLADIVOSTOK, RUSSIA, 13 - 17 JULY 2003

ABSTRACTS

PLENARY SESSION

PLANT PARASITIC AND SOIL NEMATODES

MORPHOLOGY, EMBRYOLOGY, AND EVOLUTION

MOLECULAR BIOLOGY AND DIAGNOSTICS

ENTOMOPATHOGENIC NEMATODES

ECOLOGY OF AQUATIC NEMATODES

VERTEBRATE PARASITIC NEMATODES

TAXONOMY AND SYSTEMATICS

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PLENARY SESSION

Marine biological diversity: problems, comments, methodology of researches

Andrey V. ADRIANOV

Institute of Marine Biology FEB RAS, Vladivostok, 690041, Russia, [email protected]

Terminology and six main levels of biodiversity have been suggested and discussed. Species richness and taxonomic diversity of terrestrial, freshwater and marine organisms are compared based on a new quantitative estimations. About 1.5 million of terrestrial species and 320 000 species of water organisms have been described up date. Despite more than 200 years of intensive researches biologists have described only about 280 000 marine species including about 180 000 marine invertebrates. Of the 33 phyla of Metazoa listed in the text, 31 phyla are found in the sea, 13 of which are exclusively marine. Only 17 phyla are found in the fresh water and 11 phyla are found in terrestrial environments. Only two phyla, the fresh water Micrognathozoa, and the terrestrial Onychophora, are thought to be non-marine endemics. What does it mean for forming of marine biodiversity? Marine diversity in the coral reef, coastal environments, deep-water macrobenthos and in meiofauna are calculated and discussed. Number of species for all of the metazoan phyla are quantified. Approximately 25 million undescribed macrobenthic species and about 20-30 million of meiobenthic species, including more than 10 millions of marine nematodes, are suggested in the deep-sea based on modern extrapolations. Hypothesis explaining high benthic species diversity in the deep-sea sediments are summarized. New global and regional initiatives, methodology of marine researches and biodiversity research methods are discussed.

Multikingdom system of the living things

Anatoly L. DROZDOV

Institute of Marine Biology FEB RAS, Vladivostok, 690041, Russia, [email protected]

The five-kingdom system of cellular organisms suggested in 70s (Whittaker, 1969; 1977; Whittaker, Margulis, 1979; Margulis, 1974; 1981; Margulis, Schwartz, 1982; 1988)) is in common use now. This system was primarily associated with sharp expansion of researches in microanatomy, cytology, and cytochemistry, especially those using ultramicroscopical techniques. In addition, numerous investigations were conducted in the area of phylogenetic cladistics of Hennig, genetic systematics, karyology, comparative biochemistry, immunology, and a number of other rapidly developing scientific disciplines. The five kingdoms are included into this system: three higher kingdoms of plants, fungi, and animals, and two lower kingdoms: eukaryotic Protista and prokaryotic Monera. L. Margulis clearly delineated the kingdom Protoctista from the three superior kingdoms and transferred to it all algae, all flagellate forms of fungi, and fungilike organisms. She thus made monophyletic the kingdoms of plants, fungi, and animals but increased the heterogeneity of the kingdom Protoctista. L. Margulis herself frankly admitted that "the protoctist kingdom becomes looking as if it were a dump". Retaining algae in the plant kingdom and diploid water moulds in the kingdom of fungi, she "straightforwardly admitted the polyphyletic nature of the plant and animal kingdoms". From these words of L. Margulis, it unambiguously follows that all schemes with few eukaryotic kingdoms (1 to 4) will err toward inadmissible polyphyly, as is confirmed by contemporary cytological and, especially, molecular biology data. The studying of bacteria indicated the two different groups: Eubacteria and Archaebacteria (Archae). The multikingdom system of the living things advanced by us (Kussakin, Drozdov, 1994, 1998) comprises 11 kingdoms of prokaryotes and 15 kingdoms of eukaryotes. It was based on the principle of conservatism of cellular structures formulated by V.F. Mashansky and A.L. Drozdov (1975; 1988). In these works, we made an attempt to analyze a biological paradox: at the molecular, cellular, organismic, population, and species levels, biological diversity is astonishing, while the subcellular and organoids levels are extremely conservative. There are only two major ultrastructural patterns of biological membranes and 7-8 patterns of structure of cell walls, 2-3 patterns of ribosomes, 3-4 patterns of organization of synthetic apparatus, and only six patterns of ultrastructural organization of plastids, 4-5 patterns of organization of mitochondria, 3 patterns of organization of nuclear apparatus, and 7-8 patterns of organization of kinetic apparatus. We postulated that organisms having the same structure of cellular organelles belong to the same kingdom of living organisms. "Kingdom" is an objective taxonomic category, as it is in the case with "species". We represented our system in two modus - as a table and as a drawing.

The database of class Nematoda from the Sea of Japan

Natalia P. FADEEVA*, Olga I. DASHCHENKO**

Far East State University, Vladivostok, 690600, Russia, *[email protected], **[email protected]

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The collection of free-living marine nematodes of Zoology Department of Far Eastern State University is growing since 70th. Now it has about 10 thousand individuals and more than 280 species (including type specimens) of marine nematodes from the Sea of Japan. Moreover there is material from the Far Eastern Seas, Japan, Vietnam, Indian Ocean, the Pudget Sound Bay, New Zealand and Australia. The systematic database of class Nematoda from the Sea of Japan using the Access 2000 has been created with the purpose of systematization and representation of the material in the convenient form for perception and operation. The database contains 15 tables, 10 of which represent taxa (subclass, order, suborder, family, subfamily, tribe, subtribe, genus, subgenus, species), and 5 have keys for the identification. Currently the diagnoses of 110 genera registered in the Sea of Japan are in a database. The following names of fields are presented: "A species Code", "A name of species for searching", "Species", "Description of species", "Ecological data", "Geographical distribution", "Literary data", "Male", "Female", "Types", "Material", "Figure", "Legend of figure". The name of a field "A species Code" is a counter, in case of adding a new species to the database it is automatically appropriated a numeral code. The Fields "A name of species for searching" and "Species" are text. It is possible to include to a text field the information no more than 255 symbols length. The field of "Figure" contains subjects such as OLE (drawings, EXCEL table, WORD documents etc.), in remaining fields data are situated such as MEMO. 4 elements were used for control such as: a legend, a button, a drawing and a rupture of page in the database. The database is initial stage of registration of nematofauna and bionomics of the Sea of Japan. It will be completed periodically due to new data. The database will facilitate an identification quality control to the specialists.

Phylogeny of Bursaphelenchus conicaudatus, and its vector, Psacothea hilaris

Natsumi KANZAKI*, Kazuyoshi FUTAI**

Laboratory of Environmental Mycoscience, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan, *[email protected], **[email protected]

In order to examine whether or not the co-speciation occurs in a phoretic relationship, molecular phyogenetic analysis on Bursphelenchus conicaudatus, and its phoretic host, Psacothea hilaris was conducted. The beetle is widespread in Japan, Taiwan, and the continental China, and is divided into 13 geographical subspecies. First of all, ten subspecies of the beetle were collected in various districts of Japan, and the molecular phylogenetic analysis based on the cytochrome oxidase subunit I in the mitochondrial DNA was conducted both for the nematode isolates and for its vector beetle subspecies. The analysis divided 30 isolates of nematodes obtained from 10 subspecies of beetles into five isolate groups and 10 subspecies of beetles (totally 40 individuals) into four subspecies groups, respectively. The nematode isolate groups and the beetle subspecies groups were corresponding to each other, and the mechanism of the co-speciation could be related to the paleogeography of Japan, though there were several discrepancies. Intraspecific vector replacements were supposed to occur in the isolates of B. conicaudatus on two islands. These discrepancies were found between nematode isolates and their corresponding beetle subspecies and were assumed as concomitant to the migration process of the beetle and/or the nematode populations, i.e., a beetle population and its associate nematode population on an island might have migrated from different provinces. These results suggest that 1) the co-speciation can occur in a phoretic association between the nematodes and the beetles, and 2) the co-speciation could be disturbed by extinction and re-migration of populations.

New ideas on the animal phylogeny and the position of nematodes in the system of animal kingdom

Vladimir V. MALAKHOV

Faculty of Biology, Moscow State University, Moscow, 119992, Russia, [email protected]

Molecular biology introduced new ideas into phylogenetic investigations. As a result of DNA sequence study, the new taxa were created and the animal classification changed drastically. Nematodes fell within the new superphylum taxon Ecdysozoa. In modern classification, nematodes are coupled with Arthropoda, Onychophora, Tardigrada, and Cephalorhyncha instead of previously accepted conceptions on the relation of nematodes to turbellarians, gastrotrichs, rotifers etc. The total lack of ciliary epithelia, strong cuticularization, absence of intracuticular microvilli and molting, disappearance of trochopore larvae, non-spiral type of cleavage, lack of true coelom, the absence of free nerve endings and setae-like sensilla, and other features of nematode morphology and development could be explained on the basis of the derivation from ecdysozoan ancestor. The adoption of the ecdysozoan conception makes us to assume the evolutionary reduction of limbs and segmentation in nematode ancestor as a result of burrowing habits. The evolutionary model of nematode origin could be roughly pictured as a row Onychophora-Tardigrada-Nematodes.

Parasitic Dwarf Males: Biology and Evolutionary Implications

Alexei V. RYBAKOV

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Institute of Marine Biology FEB RAS, Vladivostok, 690041, Russia, [email protected]

Dwarfs represent an extreme case of sexual dimorphism, when highly reduced males inhabit the female providing immediate fertilization of deposited eggs. They are rather widespread, being particularly common, when contacts between specimens of different sex are difficult (due to low population densities, sessile or parasitic mode of life, "patchiness" of the habitats, etc.). In these cases we could observe development of some peculiar reproductive strategies, comprising hermaphroditism, self-fertilization, or asexual reproduction. However, presence of cross-fertilization in life cycle provides obvious selective advantages, therefore long-term (or even lifetime) associations between specimens of different sex sometimes appear the most favorable, often resulting in the development of dwarf males. Dwarfism provides some important advantages. There is no need of search for breeding partners. The development of such males is much faster and the need for metamorphosis with its inherent risks is often reduced, as many dwarfs exhibit neoteny. Dwarfism also minimizes negative effects on a host that otherwise would have at least two large parasites. The associated expenses are just those of spermatogenesis, with a little more to support certain somatic growth (as the males are usually extremely reduced). Sperm production is thought to be relatively cheap, so the cost to the female for hosting even several dwarfs would be not too high. Female organism provides a habitat for such an "integrated" male and performs most of its life activities (except spermatogenesis). To host the dwarfs, females acquire some particular organs (pockets, cavities, or male receptacles of different kinds), sometimes of a complicated structure. These adaptations benefit the parasitism of the dwarf males, favoring their fast and profound degeneration. The processes associated with the development of dwarfism are very similar to those observed during the course of evolution in parasites. The phenomenon of dwarf males might be considered as a peculiar pattern of intraspecific parasitism.

Master of Science in Nematology: a success story

Nic SMOL

PINC, Vakgroep Biologie, Universiteit Gent, Ledeganckstraat, 35; B-9000, Gent, Belgium, [email protected]

The Postgraduate International Nematology Course at Ghent University has been created in 1992 to meet the needs of educating of students and scientist in Nematology. Most of the students are coming from developing countries, all over the world. The course started with 7 students and gradually increased to the present batch of 18 students, chosen out of about 90 applications. Within the past 10 years 98 students have obtained the degree of "Master of Science in Nematology". The results of the past decade are discussed and future prospectives, such as the new programme, are explained. In view of the European unification of the diplomas and the consequent introduction of the Bachelor and Master degree, new ideas for switching from one year to two years programme are discussed. Details about the admission requirements, possible scholarships and miscellaneous will be given.

Caenorhabditis elegans - 2002 Nobel prize winner

Sergei E. SPIRIDONOV

Institute of Parasitology of RAS, Leninskii pr. 33, Moscow, 117071, Russia, [email protected]

"2002 Nobel Prize for worms", i.e. presentation of the highest scientific prize for three well known specialists (S. Brenner, H.P. Horvitz, J. Sulston) in Caenorhabditis elegans developmental biology and genetics was an outstanding event of last year. Surely it was accepted with satisfaction in numerous laboratories dealing with modern developmental biology, but such a decoration is simultaneously sending very clear and powerful message to nematological community as a whole. International scientific community is paying a tribute not only to terribly profound studies in biology of single animal species, but in fact single and simple decision made more than four decades ago is pinpointed - an idea to use nematodes as biological models for developmental biology. Immanent features of nematodes and participation of elite biological workforce were probably two main factors, which produced such revolutionary results. Classical nematology was crucial to choose C. elegans as model object, with researchers like Nigon witnessing the nematode virtues. On early stages of these investigations, nematodes as transparent creatures of few hundreds of cells were studied in very straightforward way - observing the cell divisions under microscope. It means that first steps in this huge progress were made with principally the same tools and methods as those still common for classical nematology. Even morphological facts about C. elegans considered alone - apart from developmental and genetical data - are making this nematode the best morphologically studied nematode object. (Let we mention here only "curcuit scheme" - fully mapped contacts between cells in nerve system.) Basic observation - the fact of programmed cell deaths in C. elegans development - was made by John Sulston after light microscopic examination of embryonic development. Discovered developmental and morphological facts about C. elegans can be directly applied to wider scope of nematological researches. But the clear message for all the community of classical nematologists is even more important - unbiased analysis of the data obtained with quite

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common methods of nematode examination (light microscope, TEM) could bring much more features of nematode organization than it is routinely used in nematology practice.

Benthimermithidae: a review of an enigmatic marine nematode group

Alexey V. TCHESUNOV1, Dmitrii M. MILJUTIN2

1Department of Invertebrate Zoology, Faculty of Biology, Moscow State University, Moscow, 119899, Russia, [email protected]

2Federal Research Institute of Fisheries & Oceanography, Moscow, Russia, [email protected]

The family Benthimermithidae comprises marine and mainly deep-sea nematodes parasitizing at larval stages in internal organs of benthic invertebrates. Adult stages leave their hosts and dwell freely. Since benthimermithids have a modified alimentary tract devoid a mouth and normal pharynx, they obviously feed through body wall during larval parasitism and do not feed at adult stage. Larval stages possess a stylet-like structure and glandular pharynx. Adults provide some characters (cephalic and somatic setae, lateral amphids and serial supplementary organs) indicating their relationships to free-living nematodes. The family consists of three genera, two of them (Trophomera and Adenodelphis) are monotypic, whilst the third genus Benthimermis includes over 30 species. Future synonymization of Trophomera Rubzov & Platonova 1974 and Benthimermis Petter 1980 is possible. Benthimermis is a highly diversified genus, which species vary in length from 1 mm to 17 cm. Benthimermis species demonstrate different stages of fusion of male gonads from two distinct parallel testes to one united gonad and a variety of ovary types (reflexed and outstretched, telogonic and hologonic). The majority of species is described on the base of either males or females while both sexes are known for a few species only. This circumstance poses difficult taxonomic problems within the family. (RFFI grant No. 03-04-49152).

Oogenesis in free-living aquatic nematodes: ultrastructural aspects

Vladimir V. YUSHIN1, Vladimir V. MALAKHOV2

1Institute of Marine Biology FEB RAS, Vladivostok, 690041, Russia, [email protected]

2Department of Biology, Moscow State University, Moscow, 119899, Russia, [email protected]

Original data on the oogenesis in 11 species of free-living aquatic (marine and fresh water) nematodes belonging to the orders Enoplida, Mononchida, Chromadorida, Desmodorida, Monhysterida and Plectida are reviewed. Free-living aquatic nematodes have the basic pattern of oogenesis (solitary oogenesis) and vitellogenesis (authosynthesis) described in a variety of parasitic species. The vitellogenic oocytes show great synthetic activity. Voluminous nuclei have nuclear envelope with numerous nuclear pores; the nucleoplasm contains from one to several large nucleoli. Cytoplasm is filled with ribosomes, cysternae of rough endoplasmic reticulum (RER), Golgi bodies, and mitochondria. The RER is organized into stacks and whorls. The surface area of the vitellogenic oocytes is highly increased by development of numerous microvilli or another surface modifications (infoldings, outfoldings, large processes). Phagocytosis and pinocytosis, which are indicators of heterosynthetic vitellogenesis, have not been detected. The vitellogenesis results in formation of two basic types of yolk: (i) proteinaceous membrane-bound granules and (ii) lipid droplets. The special type of granules, which are enveloped by membrane but have characteristic heterogeneous matrix, was found in estuarine (Enoplida-Tripyloidina) and fresh-water (Enoplida-Tobrilina, Mononchida, Plectida) species. Analogous granules are well known in a variety of terrestrial and parasitic nematodes as the heterogeneous or shell granules. These granules are utilized during rapid egg-shell formation after fertilization. Synthesis of the shell granules correlates well with adaptation of nematodes to such an unfavourable habitat as fresh water, soil, terrestrial plants, and digestive system. (Support for Science Schools grant No. 1219.2003.4).

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PLANT PARASITIC AND SOIL NEMATODES

Dependence of vertical distribution of cyst-forming nematodes from cultivated host plants and ways of principal tilling of soil

Anatoly G. BABICH

National Agricultural University, Kiev 03041, Ukraine, [email protected]

Subsurface tillage of soil in Ukraine earlier was being implemented in the regions of insufficient moistening of steppe area. However, last time less energy-consuming technologies are applied also in the regions of non-stable and sufficient moistening of forest-steppe area and Polesya zone. The aim of research is investigation of influence of principal tilling of soil on vertical distribution of cyst-forming nematodes. It is found that localisation of cysts in the soil depends on cultivated host plants, kinds of nematodes, types of soil and ways of its tilling. Thus, on humus-scarce black soils the prevailing number of beet nematode cysts after growing up oil radish and rape was found in plow layer up to 20 cm, sugar beet -15-30 cm, and oats nematode after cultivating crops in the range of 5-15cm. On the turf and podzol soils with shallow lying of fertile layers the range of 10-20 cm was the most invaded by the beet nematode and the range 0-10 cm by oats nematode. Thus, the distribution of nematode cysts across the soil depends on depth of prevailing lying of root system of host plants. That is why their maximum number on black soils is usually situated 5-10 cm deeper than in turf-and-podzol soil. The disc tillage and flat-cut plowing cause minimal influence on vertical distribution of cysts. Plowing with soil layer turning leads to more even distribution of cysts in the soil range of 0-20 cm in case of usual plowing, and 0-30 cm in case of deep plowing for sugar beet plantation. Therefore, applying only subsurface soil tillage would promote accumulation of cyst-forming nematodes in the area of maximum location of root system of host plants. So it is advisable to apply plowing with the soil layer turning in case of technical and vegetable crops, and to apply flat-cut plowing in case of grain crops.

Soil nematodes as bioindicators for territories along main oil pipelines

Alexander S. EROSHENKO

Institute of Biology and Soil Sciences FEB RAS, Vladivostok, 690022, Russia, [email protected]

Investigations of natural biocenoses for soil nematode determination were carried out in the North Sakhalin from the north-east Piltun Bay to the north-west Pogibi Cape on the territory near to the main oil pipe in 2001. There were selected 4 ecological-faunistic complexes (EFC): i) coast-seal, ii) forestry, iii) riverside, iv) meadow-marshal. Specimens from 25 families of different groups were determined. Dominant bacterial feeders were represented by Cephalobidae and Plectidae; fungal feeders were represented by Tylenchidae. These families were main groups both frequently and domineering. Bacterial feeders with families Plectidae, Cephalobidae, Prismatolaimidae, plant feeders with Tylenchidae and root-ectoparasite with the genus Hemicycliophora (which was collected from rhizosphere of Pinus pumila) were dominant in the coast-seal EFC. The number of nematodes was limited from 2680 thousand specimens per 1 m2 of soil. Bacterial feeders with Plectidae, Cephalobidae, Prismatolaimidae, plant and fungal feeders with Tylenchidae and Diphterophoridae were dominant in the forestry EFC. The omnivorous Dorylaimida were dominant in the forestry EFC too. The nematode number was limited average 1360 thousand specimens in deciduous forests, 8035 thousand specimens in fir forests. The number of nematodes in burnt-over forests was noted as 620 thousand specimens. The same groups were dominant in the riverside EFC, and the number of nematodes registered was from 1886 to 3680 thousands specimens per 1 m2 of soil for different biotopes. The lowest density (584 thousands) was noted in the meadow-marshal EFC. The bacterial feeders Alaimidae, Prismatolaimidae, Plectidae and Teratocephalobidae were registered in this complex as dominants. Most nematode taxa had Maturity Indexes c-p 2 and c-p 3. They relate to constant inhabitants of investigative biotopes and reflex nondisturbed natural ecosystems. (Financial support - Ecological Company "Sakhalin").

Distribution of plant-nematodes in the Russian Far East

Alexander S. EROSHENKO, Tamara V.VOLKOVA

Institute of Biology and Soil Sciences FEB RAS, Vladivostok, 690022, Russia, [email protected]

The order Tylenchida includes 160, Aphelenchida - 67, and Dorylaimida - 15 species of 242 species of plant- nematodes found in plants or rhizosphere of roots in the Russian Far East. On the coniferous the main parasites from 163 species, found in rhizosphere and vegetative organs of wood, brushes and lianas plants in natural forests of Far East of Russia are species of the family Tylenchidae, genera Rotylenchus, Hemicriconemoides, Ogma, Pratylenchoides, the species Criconema varicaudata, Mesocriconema curvatum,Criconemoides sabulosus, Helicotylenchus clarkei, Hemicycliophora montana, Xiphinema thornei, Bursaphelenchus

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mucronatus. The root parasites Meloidodera sikhotealiniensis, Meloinema maritima, Sphaeronema salicis are widespread in broadleaves trees in valley forests. 109 species of nematodes are parasites on grass plants. Group of spiral nematodes of genera Helicotylenchus and Scutellonema and criconematids (Criconema, Mesocriconema and Ogma) have a wide spectrum of host-plants and may be considered as dominant species. Cyst-forming nematode Globodera artemisiae (sibling species of potato globoder) is widely distributed in the Far East of Russia and prefers Artemisia spp. Other species of cyst-forming nematodes: Heterodera graminis, H riparia, Afenestrata orientale parasitize on Poaceae in Primorye Territory, Heterodera humuli - on Urtica sp. in Kamchatka Region. Anguina agrostis is widespread in all regions and forms galls on the different species of Poaceae. Subanguina graminophila and S.radicicola inhabit small-reed (Calamagrostis langsdorffii) in Primorye, Khabarovsk and Sakhalin Regions and form leaf and root galls respectively. Anguina moxai forms leaf galls on Artemisia spp. and is widespread in the south of Primorye Territory. 10 species from 20 plant-nematodes are known as parasites of soybean roots. Cyst-forming nematode Heterodera glycines (common species in Primorye and Amur Regions) is most serious parasite of soybean. 3 pathogenic nematodes species are registered on the potato: cyst-forming nematode Heterodera rostochiensis (localy spreading in personal plots of citizens in all regions of Far East of Russia), Ditylenchus destructor and D dipsaci. Two species of gall-gorming nematodes: stem nematode Ditylenchus dipsaci and root endoparasite Pratylenchus vulnus (parasitizing on different vegetable and floral cultures in greenhouses) are the most dangerous for vegetable cultures. Aphelenchoides fragaria and Ditylenchus dipsaci damage strawberry in all Regions. Aphelenchoides besseyi is parasite of rice, and Heterodera graminis is parasite of maize in Primorye Region.

The role of culture medium in the tolerance of soil nematode Cephalobus persegnis to lead acetate effect

Anatoly I. GRUZDEV, Ludmila I. GRUZDEVA, Tatiana E. KOVALENKO, Elizaveta M. MATVEEVA

Institute of Biology, Karelian Research Centre, RAS, Petrozavodsk, 185610, Karelia, Russia, [email protected]

Cephalobus persegnis was grown on the 2 % standard bactoagar "Difco" (BA) and potato dextrose agar (PDA) with fungus Althernaria tenuis. Activities of cytochrome C oxidase, aldolase and NAD-linked malate dehydrogenase (MDH) isozymes were measured to determine the physiological state of C. persegnis exposed to lead acetate at concentrations from 0.75 to 25 mg/l for three days. It was displayed that in the C. persegnis cultured on BA without lead acetate the activities of the aldolase and the MDH-A2 isozyme (gluconeogenic enzymes) were significantly lower than those of C. persegnis cultured on PDA. It gave an indication of the gluconeogenesis weakening, admittedly associated with glucose synthesis from galactose which is the major constituent of BA. Experimentally, in C. persegnis cultured on BA, the aldolase activity grew progressively up twofold whereas the lead acetate dosage increased. It could be indicative of that storage carbohydrates in C. persegnis broke down by glycolysis. This assumption was confirmed by the correlation analysis applied to the data received. Using the method of probabilistic estimation of the toxic effect (Kopanev et al., 1988), we have found that the minimal toxic concentrations of lead acetate for C. persegnis cultured on PDA and BA were 0.141 ± 0.025 and 0.045 ± 0.019 mg/l, accordingly, and differed significantly (p = 0.025). Thus, C. persegnis cultured on BA differed from that cultured on PDA in fermentative status and this could be the reason why they had a reduced vitality observed on exposure to lead acetate.

The nematodes of family Tylenchidae Orley, 1880 in the Russian Far East

Inna P. KAZACHENKO

Institute of Biology and Soil Sciences FEB RAS, Vladivostok 690022, Russia, [email protected]

Representatives of 5 subfamilies, 9 genera and 21 species of family Tylenchidae were registered in the Russian Far East. Specimens from genera Filenchus (F. retusus, F. thornei, F. vulgaris, F. hamatus, F. ditissimus), Coslenchus (C. costatus), Malenchus (M. bryophilus, M.pressulus), Cephalenchus (C.hexalineatus, C. leptus) are widely distributed. Filenchus helenae and Tylenchus elegans are registered very seldom. Tylenchus davainei and Aglenchus ainakamure are revealed on Sakhalin only. Coslenhus bisexualis is noted in the Russian Far East at first. It was known earlier for Indian nematode fauna. Basiria paraobliqua is widely distributed in Primorye Region. Basiria graminophila and B. tumida are found seldom. Malenchus bryanti is collected from forest trees as single specimen. Lelenchus leptosoma is noted on Sakhalin and Primorye Regions.

Local and Systemic induced resistance to the root-knot nematodes (Meloidogyne incognita) in tomato by different doses of chemicals and its comparison with biological control agent (Paenibacillus polymyxa)

Hafeez U. KHAN1, Young Ho KIM2

1Pakistan Science Foundation, Islamabad, Pakistan

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2School of Agricultural Biotechnology, Seoul National University, Suwon 441-744, Korea

A 60 days glasshouse experiment was conducted to assess the influence of two doses of BABA (DL-B- aminobutyric acid) and BTH (Benzo-(1.2.3)-thiadiazole-7-carbothioic acid S-methylester) with 20 mM and 40 mM and one dose of GBR-I (Paenibacillus polymyxa) on the multiplication of Meloidogyne incognita and growth of tomato (Lycopersicon esculentum). GBR-I was better at improving tomato growth and reducing number of galls formed on each tomato seedlings as compared to BABA and BTH. Study revealed that GBR-I gave minimum number of galls (9.2) on each tomato seedlings which was 91.8 percent less as compared with the control. BABA with 40 mM dose ranked second in performance as it gave 17.6 galls per plant which was 84.3 percent less as compared to control. The minimum reduction in gall formation on each tomato seedling was observed with BTH at 20mM dose and it was only 62 percent less than control. Thus biological control treatment with GBR-I (Paenibacillus polymyxa) proved best in reducing the number of galls formed on each tomato seedlings and improving the plant growth as compared to chemical treatments with BABA and BTH.

Discovery of the nematode Sheraphelenchus sp. in dead-wood of the pine Pinus koraiensis from Primorski territory, Far East Russia

Irina A. KRUGLIK

Institute of Biology and Soil Sciences FEB RAS, Vladivostok, 690022, Russia, [email protected]

The genus Sheraphelenchus Nickle, 1970 includes only two species. They have a phoretic relationship with the nitidulid beetles; the free-living generations were collected from split and decaying oranges in USA (Texas, California, Hawaii, Colorado and New Mexico) and in Italy. Females of the genus Sheraphelenchus were collected from the dead-wood of the pine Pinus koraiensis in Primorsky Territory, Far East of Russia. The species from Primorye differs from both known species by morphological characters. It is characterized by long body (1,27-1,8 mm against 0,48-1,19 mm), long stylet (16-17 μm & 12 μm), location of nerve ring, long oesophageal glands. The Primorye females differ from one of the type species, S. entomophagus Nickle, 1960, by length and shape of tale.

Distribution of nematodes inside trunk and branches of the dead 200 years old pine Pinus koraiensis

Irina A. KRUGLIK

Institute of Biology and Soil Sciences FEB RAS, Vladivostok, 690022, Russia, [email protected]

Primary location of different nematodes in separate parts of the stem and branches was found during investigation of the nematodefauna of dead-wood of the pine Pinus koraiensis. Most nematodes collected were mycophagous and saprobiont forms. Content of the nematodes in the bark was approximately: 50% - Cryptaphelenchus ipinius; 30% - Parasitorhabditis acanthocini, 22% - Bursaphelenchus mucronatus and 8% - Ektaphelenchoides pini. Specimens of Diastolaimus grossus were more than half of all nematodes in the branches; Panagrobelus phloesini, Aphelenchoides saprophilus, Bursaphelenchus sp. and Deladenus sp. were collected in uniform proportions. The species B. mucronatus occupies a core of the middle part of the stem where a funous lesion was not visualized. Its quantity in this sample was estimated as 97,2%. Considerable fungous infection was found in the external layer of the middle part of the stem. The dominant species in this habitat were Bealius pissodi - 55%, P.acanthocini - 19%, Rhabditis cucumeris - 12%, Neoditylenchus corniculatus - 10 % and B. mucronatus - 4 %. Areas with considerable fungous infection were not observed in the upper part of the stem while fungi were scattered in sawcut almost evenly. Here the estimated quantity of B.mucronatus was 71%. C. ipinius dominated (52%) in the lower part of the stem with strong fungous infection. The species Bursaphelenchus silvestris, Neodiplogaster wacheki, Parasitorhabditis acanthocini, Neoditylenchus corniculatus and Parasitaphelenchus sp. were also registered here.

Fauna of xylobiont nematodes of the pine Pinus densiflora from Primorye

Irina A. KRUGLIK

Institute of Biology and Soil Sciences FEB RAS, Vladivostok, 690022, Russia, [email protected]

Fauna of xylobiont nematodes of the pine Pinus densiflora is characterized by much less number of species compared with other pine species growing in Primorye. Quantity of nematodes is also much less. Nematodes were not found almost in half of trees investigated. 18 species of nematodes were found in stems and branches of a pine Pinus densiflora: Plectus rhizophilus, Macrolaimus arboreus, Diastolaimus grossus, Heterocephalobus elongatus, H. cf. pauciannulatus, Panagrolaimus rigidus, Panagrobelus phloesini, Protodiplogasteroides saperda, Ditylenchus cf. tenuidens, D. uniformis, Deladenus durus, Aphelenchoides hamatus. A.paramonovi, A.rhytium, A.saprophilus, Ektaphelenchus prolobus, Bursaphelenchus mucronatus. Only two of them (H. elongatus and E. prolobus) were not found in other pine species. Most of nematodes

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collected are mycophagous and saprophagous forms. More abundant saprophags were P. phloesini, M. arboreus and D. grossus. A. saprophilus was more abundant mycophag. B. mucronatus, always accompanied by other species of nematodes, was found in 12,5% of the investigated trees. Such frequent occurrence is much more higher than in the forest P.koraiensis but less than in P.koraiensis and P.silvestris from the forest nurseries. Quantity of B. mucronatus in wood of P. densiflora is very low. This fact testifies against the pathogenicity of B. mucronatus for P. densiflora.

Review of nematode order Aphelenchida Siddiqi, 1980 from stems and branches of pines from Primorye

Irina A. KRUGLIK

Institute of Biology and Soil Sciences FEB RAS, Vladivostok, 690022, Russia, [email protected]

Nematodes belonging to the order Aphelenchida are very numerous in dead-wood of pines in Primorye. 28 species of aphelenchids belonging to 10 genera of five families (Paraphelenchidae, Aphelenchoididae, Seinuridae, Ektaphelenchidae, Parasitaphelenchidae) were registered. The family Aphelenchoididae is represented by three genera (Aphelenchoides, Laimaphelenchus, Sheraphelenchus) with 11 species. Genus Sheraphelenchus was found in Russia for the first time. Most of the nematodes found belong to the genus Aphelenchoides - 9 species. One of these species is new for science; five species were found in the Russian Far East for the first time. Family Parasitaphelenchidae is represented by two genera (Bursaphelenchus, Parasitaphelenchus) with 10 species. Seven species belong to the genus Bursaphelenchus. Three of them are new for science; two species are registered in the Russian Far East for the first time. Three species from the genus Parasitaphelenchus also were found in the Russian Far East for the first time. Three genera (Ektaphelenchus with 3 species, Cryptaphelenchus with 1 species and Ektaphelenchoides with 1 species) belong to the family Ektaphelenchidae. All five species were registered in the Far East for the first time. Species from the families Paraphelenchidae and Seinuridae were found in the dead-wood of pines only as sporadic specimens. One species was registered for each family. Thus, four of 28 species of aphelenchids found in the Russian Far East are new for science; 16 species were found for the first time, eight species have been registered previously.

Description of Meloidogyne zhanjiangensis n.sp. from China

Jinling LIAO*, Zhixin FENG

Lab of Plant Nematology, College of Environmental and Natrual Resource, South China Agricultural University, Guangzhou 510642, P.R.China, *[email protected]

A new species Meloidogyne zhanjiangensis on pea from Guangdong province, China was observed and described. For female: stylet 12-13.3 μm length; perineal pattern changed much, near round, sometimes regular; arch low to middle high; fine striae, smooth to waved; tail terminus covered with irregular circles. In compare with M. arenaria, M. zhanjiangensis has irregular pattern, perineal pattern with higher arch, smooth striae in lateral field, irregular former lip, whereas M. arenaria has typical dumbbell lips. Lower esterase migrating rate belongs to VS1-S1 type. Nal type of malic dehydrogenase. For the second stage juvenile: body length 320-475μm; stylet 13.5-15 μm length; tail has 1-4 indentations, small rounded terminus. Smaller mtDNA fragment compared with M.arenaria (0.7 kb for M. zhanjiangensis, 1 kb for M. arenaria). The host reaction of Meloidogyne zhanjiangensis on some crops also was discussed (Natural Science Foundation of China, grant No. 30170611).

Structure of soil nematode communities in potato fields infested with potato cyst nematode (PCN)

Elizaveta M. MATVEEVA, Ludmila I. GRUZDEVA, Tatiana E. KOVALENKO

Institute of Biology, Karelian Research Centre, RAS, Petrozavodsk, 185610, Karelia, Russia, [email protected]

Plant parasitic nematode complexes are usually formed from natural communities existing in local areas. Agroecosystems are characterized with monoculture that enhances a distribution of specialized plant parasitic nematodes, once occurred in the soil (e.g. potato - PCN as example). Diversity of soil nematode fauna, eco-trophic structure of the nematode communities in the infested potato fields were compared with those of health fields and after the applying of control measures against PCN. Nematode fauna diversity was decreased in the highly infested potato fields (over 200 viable PCN juveniles per 100 g of soil): 17 genera are observed while from 22 up to 30 nematode genera were found in the slightly infested fields. Maturity index (MI) of the nematodes communities was low (2.4 - 2.5); plant parasite index (PPI) was high (3.0 -3.2). Nematodes - bacteriotrophs were the most numerous among eco-trophic groups. Plant parasites were observed as a subdominant group, and genus Globodera was the most abundant in the plant parasite complex. Polytrophs and predacious nematodes were scanty. PCN population showed the 3-fold decline after the growing of potato cultivars resistant to the nematode and a 5-fold decline under fallow for 2 years. Simultaneously, changes in the

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trophic structure of the nematode communities were established. Bacteriotrophs were still dominant; polytrophs and predacious nematodes occurred as subdominants instead of plant parasites. Changes in the plant parasite complex were also found: representatives of Tylenchorhynchus, Pratylenchus, Paratylenchus were increased in numbers. This was mainly due to a succession of plants growing on the former fields and, consequently, occurrence of new relations in agrosystem. (Grant from Department of General Biology Programme "Biological resources").

Response of a Root-Knot Nematode, Meloidogyne incognita, to Homobrassinolide and Epibrassinolide

Puja OHRI1, Satwinder K. SOHAL1, Renu BHARDWAJ2

1Department of Zoology, Guru Nanak Dev University, Amritsar, India, [email protected]

2Department of Botanical and Environmental Sciences, Guru Nanak Dev University, Amritsar, India, [email protected]

The regulatory effects of Brassinosteroids (BS) have been extensively demonstrated in plants and have been documented on animal systems too. Since plant-parasitic nematodes are intimately associated with plants for their nutritional requirement for sterol, therefore the present study was a preliminary step in this regard to see the effect of two most potent brassinosteroids - Epibrassinolide (EBl) and Homobrassinolide (HBl) on a root - knot nematode, Meloidogyne incognita. Six concentrations of both the steroids were prepared in distilled water (10-5,10-6, 10-7,10-8,10-9and10-10 M). The egg masses of M. incognita were exposed in vitro to the above six concentrations for 7 days. Observations recorded after 24, 48, 72, 96h and 7 days of hatching revealed much higher percentage of hatching in treated egg masses (except in 10-10 M HBl) as compared to control. Some higher concentrations resulted into almost double percentage of egg hatching. The second stage juveniles were also tested with the same concentrations of HBl and EBl for their vigour. Juveniles apparently were as active as in controls. The juvenile mortality in treatments was not significantly different from control.

Soil nematodes of broad-leaved wood in southern Primorye Region

Tamara V.VOLKOVA, Alexander S.EROSHENKO, Inna P.KAZACHENKO

Institute of Biology and Soil Sciences FEB RAS, Vladivostok, 690022, Russia, [email protected]

The samples were collected in broad-leaved wood where the oak Quercus mongolica is a dominant. Representatives of 26 families of soil nematodes were determined. Ecological-taxonomic analysis of nematode species showed that fauna of plant and soil nematodes is typical for natural forest cenosis. Plant nematodes are represented by seven families, where Rotylenchus feroxis and Xenocriconemella macrodora are dominant. Fungal feeders are represented by genera Filenchus, Aphelenchus and Aphelenchoides. Bacterial feeders are represented by rhabditids, cephalobids and plectids. Significant percent of fauna is formed by predators from the family Mononchidae, with the dominant genus Coomansus, and omnivores from the order Dorylaimida. The highest density of soil nematodes is noted in oak woods in the region of the lake Lotos and in Popov Island, the lowest - near the dust-heap of Vladivostok City. The highest diversity of nematode species is noted in oak woods in Popov Island, settlement Rybachii and Shamora bay, the lowest - in the region of the City’s dust-heap. Dynamics of development of nematofauna is wave-like during the vegetation season. Bacterial feeders and predators have stable dynamics type. Plant nematodes have more sharp seasonal fluctuations of the number. Main peak of the number of criconematids was accounted in June and September, for spiral hoplolaimids - at July and October. Sharp reduction of abundance of these groups was observed at August. It seems, this is connected with aridity of this month. The omnivorous form of Dorylaimida had practically constant, relatively high number from May to July, sharp reduction in August and September and an increasing number in October.

The influence of the soil acidity (pH) on nematode density

Tamara V. VOLKOVA, Alexander S. EROSHENKO, Serafima V. TEPLYAKOVA

Institute of Biology and Soil Sciences FEB RAS, Vladivostok, 690022, Russia, [email protected]

The soil samples were collected from rhizosphere of oak Quercus mongolica for studing the influence of the soil acidity on nematode density in August-September, 2001. Sampling sites were disposed along the highway Vladivostok-Artem nearby the Ussuriisky Bay at the distance 2 km each from other including ash disposal area and cities’ dust-heaps - technogenic and urban pollution zones. The highest acidity was revealed in the region of dust-heap and neighbouring sites (pH 3.1-3.8). There was marked the reduction of nematode density compared with other sites (N=245 per 1000 ml soil). The bacterial feeders were most abundant from the trophic groups. This group was represented by rhabditids, cephalobids and plectids. The lowest acidity, near to neutral, was revealed in the region of ash disposal and neighbouring sites (pH 5.5-7.0). The highest nematode density (N = 807 per 1000 ml soil) were recorded here. Main peak of the number was accounted among the

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fungal feeders. This group was represented by the genus Filenchus. The soil acidity within the limits of pH 4.0-6.5 was noted for other sites. The optimal soil acidity (pH 5.1-5.5) was recorded for the plant feeders with the dominant species Rotylenchus feroxcis and Xenocriconemella macrodora. Other nematode groups responded to pH insignificantly.

The dependence of nematode density from heavy metal content

Tamara V. VOLKOVA, Alexander S. EROSHENKO, Serafima V. TEPLYAKOVA

Institute of Biology and Soil Sciences FEB RAS, Vladivostok, 690022, Russia, [email protected]

The 35 soil samples were collected from rhizosphere of oak Quercus mongolica for study the dependence of nematode density on heavy metal content in August-September, 2001. Sampling sites were disposed along the highway Vladivostok-Artem nearby the Ussuriisky Bay at distance 2-km each from other. Heavy metals were represented by Fe, Mn, Zn, Cu, Co, Ni, Cd, Pb. Content of general and mobile forms of heavy metals was revealed in the collected samples. General form of heavy metal content showed stability of these elements. Dependence of nematode density from general form of heavy metal content has not found. This form reflects only the general pollution level. Changing of nematode density was evaluated by mobile form of heavy metal content. The constant nematode density of all groups depended from pollution stability. Small nematode density deflections were recorded during sharp overranging of admissible concentration limit (ACL). The high ACL was noted in 6 samples only. The decrease nematode density except predators was noted during overranging of the lead (Pb) content. Decrease of density of all nematode groups was noted during the overranging of cadmium (Cd) content. Combinations of overranging lead + copper (Cu) and lead + zink content were revealed the regularity of plant and fungal feeders density increase. Probably, zinc and copper smooth out negative activity of lead and cadmium, but to certain degree.

Research of intensity of Globodera rostochiensis infection in potatoes with endotoxine-gene Bacillus thuringiensis

Svetlana V. ZINOVIEVA1, Zhanna V. UDALOVA1, Olga L. OZERETSKOVSKAYA2

1Institute of Parasitology RAS, Moscow 119071, Russia, [email protected], [email protected]

2Bach Institute of Biochemistry RAS, Moscow 119071, Russia, [email protected]

Transgenic plants with expressed modified genes of entomocide proteins, encoding only their toxic domens, are known to exhibit unexpected resistance spectrum to other pathogens. The greatest progress in this field was achieved with transfer to plants of endotoxine gene Bacillus thuringiensis which could inhibit not only insects but other invertebrates, including nematodes. Effect of cyst-forming nematode Globodera rostochiensis (race 3) on potatoes with endotoxine-gene B. thurigiensis var. tenebrionis (Bt) developed on Superior ("Monsanto" USA) cv., resistant to Colorado potato beetle, was studied. Plants with unchanged genotype susceptible to G.rostochiensis were used as controls. The experiments were conducted in greenhouses. Potatoes were infected at germination with 10 000 larvae/plant. Fifteen replicates were completed for each variant. G.rostochiensis cyst counts were made in 2 months in roots and soil after germination. The results showed that cyst number in roots of transgenic potatoes was 2.3 times as low as in control. The number of larvae presented in cysts did not differ in both variants. Transgenic potatoes with endotoxine-gene Bt were more resistant to phytopathogenes in treatment with biogenic elicitors.

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MORPHOLOGY, EMBRYOLOGY, AND EVOLUTION

Comparative study of the female reproductive system within the Tylenchomorpha (Nematoda: Tylenchina)

Wim BERT, Ruben VAN GANSBEKE, Rita VAN DRIESSCHE, Myriam CLAEYS, Etienne GERAERT and Gaлtan BORGONIE

Ghent University, Department of Biology, Nematology section, Ghent, [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

The cellular gonoduct structure of 97 species belonging to the Tylenchomorpha and of eight species belonging to outgroups were studied to explore diagnostic applications of gonoduct characteristics and their use to assess phylogenetic relationships within the Tylenchomorpha. The morphology of the spermatheca has been shown to have particular value as a taxonomic character, while the cellular arrangement of oviduct and uterus seems to be a reliable phylogenetic marker. The gonoduct of the studied tylenchs (without Criconematoidea) displays important similarities: the oviduct consists of two rows of three to seven cells; the spermatheca comprises 10 to 20 cells and the uterus cells are arranged in three or four rows. The spermatheca of Meloidogyne is formed by a variable number of thick, lobe-like cells making it distinctive from any other known nematode genus. Ultrastructural data substantiate that the Meloidogyne spermatheca and oviduct are exceedingly complex structures and that their function is broader than a simple conduit for oocytes and receptacle for sperm. In the Criconematoidea, cells of the uterus-spermatheca complex are not arranged in distinct rows and a lumen is present. The gonoduct structures of the Aphelenchidae and Aphelenchoididae are remarkably distinct from each other and from the remaining Tylenchomorpha, that does not preclude the polyphyly of the Aphelenchoidea as suggested by molecular data.

Peculiarities of the life-cycle of the free-living marine nematode Oncholaimium ramosum (Enoplida: Oncholaimidae) from the Sea of Japan

Inna L. DAVYDKOVA, Natalia P. FADEEVA

Far East State University, Vladivostok 690600, Russia [email protected]

The free-living marine nematode Oncholaimium ramosum Smolanko et Belogurov, 1987 dominates in the sediments of chronic harbour pollution of the Far Eastern ports. Study of life cycle was carried out both in laboratory conditions during two years and using the season data. Observations on the embryonic and postembryonic development, and also the moulting processes are presented. 35 egg layings, 400 eggs and about 1000 nematodes of this species are investigated. Nematode O. ramosum have two generations in the Sea of Japan. The egg deposition continues from February to July (spring generation) and from September to November (autumn generation). 100% mortality of female is registered afrer completion of the egg deposition. The females Oncholaimium ramosum are observed from egg deposition to death almost continuously. The female deposit consists from either clusters of 2-38 eggs or from a single egg. There are line, bunch or lump layings. Ellipsoid eggs are 95 - 120 μm long and 50 - 94 wide. The cleavage of 43-50% of eggs in a laying is observed several right after deposition. The others 50% of eggs lag behind in development for 8-10 days. The process of hatching from initial to last larva in laying proceeds also 8-10 days. The stages of 2, 4, 8 blastomeres, late gastrula, and vermiform larva were described. The average time of development of Oncholaimium ramosum from start of cleavage to hatching is 1-1,4 months. The average time of hatching is 5-10 minutes. The postembryonic development includes four moulting cycles resulting in four juvenile stage designated as J1, J2, J3, and J4. The size and time of development of juveniles J1-J4 for Oncholaimium ramosum are determined. (RFBR grant No. 03-04-49573).

The cuticle structure of Tricoma sp.(Nematoda: Desmoscolecida)

Shota V. KOVALYEV

Department of Invertebrate Zoology, Faculty of Biology, Moscow State University, Moscow, 119899, Russia, [email protected]

Todate nematodes of the order Desmoscolecida remain poorly investigated with transmission electron microscope (TEM). The study is an attempt to deepen understanding of organization of desmoscolecids. The structure of cuticle of desmoscolecid Tricoma sp. from the sublittoral zone of Kandalaksha Bay of the White Sea is investigated with TEM. The cuticle is coarsely annulated and consists of 38 annuli (desms) joined by arthrodial cuticle. Desms are covered with chaotically situated plates. Three-layered epicuticle and the light homogenous layer are distinguished in transversal sections. Two layers, subsurface and inner, are observed

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under the epicuticle in longitudinal sections. The subsurface layer is light, homogenous, without any obvious features of radial striation, it presents both in desms and interzones between them. The inner layer presents only in arthrodial cuticle of interzones; it is divided into 4-5 distinct layers formed by alternating light and dark plots. Electron-dense plates and bundles of fibres lie in hypoderm under the arthrodial cuticle. Hypoderm penetrates each cuticle annulus where it underlies the homogenous layer. Body surface, head and tail regions were studied in Tricoma albimaris with scanning microscope. The pattern of head sensilla "6+4" was detected. Additional TEM data on the morphology of sensory organs of the anterior end are also given.

On the taxonomy and ultrastructure of Microlaimidae (marine Chromadoria)

Shota V. KOVALYEV*, Alexei V. TCHESUNOV**

Department of Invertebrate Zoology, Faculty of Biology, Moscow State University, Moscow, 119899, Russia, *[email protected],**[email protected]

Taxonomy of the family is revised. Original keys for definition of the genera as well as of the species within each genus (with the exception of Microlaimus) are made up. The following genera are accepted as valid: Acanthomicrolaimus, Bathynox, Bolbolaimus, Caligocanna, Ixonema, Microlaimus, Pseudomicrolaimus, Spirobolbolaimus. The genus Calomicrolaimus Lorenzen 1976 is considered to be a junior synonym of Microlaimus De Man 1880. Five species from the White, Caribbean seas and the Pacific Ocean are described as new to science: Microlaimus sp.1, M. sp.2, M. sp.3, Aponema sp.1, Caligocanna sp.1; redescriptions of 5 known species are also given. The ultrastructure of cuticle, hypoderm, musculature, intestine is investigated in Microlaimus honestus with TEM. The cuticle consists of four layers of typical structure: three-layer epicuticle, radial-striated exocuticle, meso- and endocuticle subdivided into several layers. Hypoderm is seven-cellular in cross section, with prominent lateral and ventral chords. Ventral nerve trunk is well developed and located in corresponding chord. Musculature is of polymyarian type (22 cells in the field of cross section), with contractile zone in the basal part of muscular cells. The body cavity looks as narrow spaces between internal organs. Intestine is oligocellular, with microvilli turned to the lumen.

Self-moving spermatophores: spermatozoan dimorphism in Steinernema (Steinernematidae, Rhabditida)

Vladimir V. YUSHIN1, Mutsuhiro YOSHIDA2, Sergei E. SPIRIDONOV3

1Institute of Marine Biology FEB RAS, Vladivostok 690041, Russia, [email protected]

2National Institute for Agro-Environmental Sciences, Kannondai Tsukuba, Ibaraki, Japan [email protected]

3Institute of Parasitology of RAS, Leninskii pr. 33, Moscow, 117071, Russia, [email protected]

The genus of the enthomopathogenic nematodes Steinernema includes the complex of species whose testes produce relatively small number of giant spermatozoa 50-100 µm in diameter (megaspermatozoa). The megaspermatozoa possess a set of cytoplasm features usual for common spermatozoa of the steinernematids (S. intermedia, S. feltiae) and other rhabditids. The megaspermatozoa from the testes are unpolarized cells, which contain mitochondria and numerous aberrant components known as the membranous organelles (MOs). Numerous minute spermatozoa about 2 μm in diameter (microspermatozoa) cover the surface of the megaspermatozoa. The microspermatozoa contain the same organelle set, which includes several mitochondria and the sole MO. The megaspermatozoa from the uterus have large pseudopod, the MOs are attached to the plasmalemma and are open to the exterior via pores. The microspermatozoa from the uterus were found both on the surface of the megaspermatozoa and as free cells in the uterus lumen. The MO of microspermatozoon is attached to the plasmalemma and is open to the exterior via pore. Thus, the spermatogenesis in S. tami results in the formation of spermatophores where one hypertrophied germ cell (megaspermatozoon) functions as a transportation device for numerous simplified male gametes (microspermatozoa). The megaspermatozoa from the uterus are motile cells, therefore the male gamete complex of S. tami may be defined as a "self-moving spermatophore". The uniform (5 μm in diameter) "normal" spermatozoa of some steinernematids (S. intermedia, S. feltiae) are capable of association into stable self-moving chains. This feature may be considered as a preadaptation to the formation of spermatophores. (Grant of Far East Branch of RAS for V.V. Yushin, Support for Science Schools grant No. 1219.2003.4).

Electron microscopic study of the spermatogenesis in free-living marine nematode Paracyatholaimus pugettensis (Chromadorida, Cyatholaimidae)

Julia K. ZOGRAF1, Vladimir V. YUSHIN2

1Department of Zoology, Academy of Ecology, Marine Biology and Biotechnology of Far East State

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University, Vladivostok, 690000, Russia, [email protected]

2Institute of Marine Biology FEB RAS, Vladivostok, 690041, Russia, [email protected]

Spermatogenesis and structure of mature spermatozoa in a free-living nematode Paracyatholaimus pugettensis were studied using electron microscopy. The cytoplasm of spermatocytes contains many Golgi bodies, cisternae of the rough endoplasmic reticulum (RER), ribosomes, mitochondria, and fibrous bodies (FBs). In the main cell body (MCB) of spermatids FBs and mitochondria surround the centrally located nucleus devoid of nuclear envelope; the sperm plasmalemma develops many folopodia. Narrow cellular bridge unites the MCB with the residual body which includes the synthetic apparatus of the spermatid. The immature sperm resembles in structure the MCB of the late spermatids. Mature spermatozoa from the uterus consists of the MCB and prominent pseudopodia filled with cytoskeleton filaments. The MCB contains nucleus, mitochondria, remnants of FBs, large osmiophilic bodies and vesicles with flocculent material. The spermatozoa of P. pugettensis exhibit the main ultrastructural features characteristic for nematodes: amoeboid nature and absence of the axoneme, acrosome and nuclear envelope. However, in P. pugettensis and other chromadorids studied the aberrant membranous organelles (MOs), which are typical of the nematode sperm, were not found at any stage of spermatogenesis. In this respect, its spermatogenesis differs from spermatogenesis in monhysterids and secernents but resemles one of tylenchids (Grant of Far East Branch of RAS for V.V. Yushin, Support for Science Schools grant No. 1219.2003.4).

Ultrastructure of spermatozoa in the free-living marine nematode Paracanthonchus macrodon (Nematoda, Chromadorida)

Julia K. ZOGRAF1, Vladimir V. YUSHIN2

1Department of Zoology, Academy of Ecology, Marine Biology and Biotechnology of the Far Eastern State University, Vladivostok, 690000, Russia, [email protected]

2Institute of Marine Biology FEB RAS, Vladivostok, 690041, Russia, [email protected]

The spermatozoa from testes and uteri of the free-living marine nematode Paracanthonchus macrodon (Cyatholaimidae, Chromadorida) were studied electron-microscopically. The spermatozoa from the testes are unpolarized cells covered by numerous filopodia. The sperm nuclei have oval or bean-like outlines and are surrounded by mitochondria. The peripheral cytoplasm is devoid of organelles and contains evenly distributed filamentous material. The filopodium content is the same as of the sperm cytoplasm. A submembrane layer of 13-16 nm thick longitudinally oriented microtubule-like fibres (MLF) strengthens the filopodia. The spermatozoa from the uterus are amoeboid cells with the discrete nucleus surrounded by mitochondria. The bulk of the sperm cytoplasm lacks organelles and is filled with MLF, which also underlie the sperm plasmalemma. The spermatozoa develop large pseudopods, which form the hemidesmosome-like junctions with the uterus wall. A layer of a subsurface osmiophilic material concentrating at the junction point is associated with the MLF running out into the cytoplasm as parallel arrays. The aberrant cytoplasm components of the nematode spermatozoa, namely the membranous organelles and fibrous bodies, were not found in the spermatozoa of P. macrodon. This pattern of the spermatozoon structure is interpreted as a highly reduced condition. (Grant of Far East Branch of RAS for V.V. Yushin, Support for Science Schools grant No. 1219.2003.4).

Ultrastructural study of sperm development in the free-living marine nematode Halichoanolaimus sonorus (Chromadorida, Selachinematidae)

Julia K. ZOGRAF1, Vladimir V. YUSHIN2

1Department of Zoology, Academy of Ecology, Marine Biology and Biotechnology of Far Eastern State University, Vladivostok, 690000, Russia, [email protected]

2Institute of Marine Biology FEB RAS, Vladivostok, 690041, Russia, [email protected]

Spermatogenesis and structure of mature spermatozoa in the free-living marine nematode Halichoanolaimus sonorus were studied using electron microscopy. The cytoplasm of spermatocytes contains many Golgi bodies, cisternae of the rough endoplasmic reticulum (RER), ribosomes and mitochondria. The main cell body (MCB) of spermatid contains the centrally located nucleus devoid of nuclear envelope, mitochondria and dense fibrous bodies (FBs). The surface of the spermatid develops numerous short filopodia. The Golgi bodies, cisternae of RER, and ribosomes are concentrated in the residual body of the spermatid. The immature sperm from testes resembles the MCB of spermatid. The spermatozoa from uteri lack usual postinsemination transformations. Their FBs remain intact, the surface filopodia transform into small lamellipodia. The large pseudopod found in many other nematode spermatozoa is absent. The spermatozoa of H. sonorus also lack membranous organelles (MOs), another characteristic feature of the aberrant nematode spermatozoa. The unique pattern of spermatozoa

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with FBs never associating with the MOs differs H. sonorus from many nematodes studied but unites the chromadorids and the tylenchids (Tylenchida). This conclusion is supported by filopodial nature of the sperm surface demonstrated by both taxa. (Grant of Far East Branch of RAS for V.V. Yushin, Support for Science Schools grant No. 1219.2003.4).

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MOLECULAR BIOLOGY AND DIAGNOSTICS

Survey of Russian conifers for Bursaphelenchus species and their identification using PCR-RFLPs and sequences of ribosomal DNA.

Oleg A. KULINICH1*, Sergei A. SUBBOTIN1, Pavel V. TOULDOUKOV1, Maurice MOENS2, Andrei O. REMEZOV3

1Institute of Parasitology of Russian Academy of Sciences, Leninskii Prospect, 33, Moscow 119071, Russia,*[email protected]

2Crop Protection Department, Agricultural Research Centre, Burg, Van Gansberghelaan 96, 9820 Merelbeke, Belgium

3Plant Quarantine State Inspection of Altai region, Gorkii str. 4, Barnaul, 656056, Russia

The genus Bursaphelenchus contains nearly 60 species and includes quarantine and other plant-pathogenic nematodes. Eleven Bursaphelenchus species were found in Russia. The data presented here are from a survey of conifers in the Asian part of Russia for pathogenic nematodes of the genus Bursaphelenchus. The analysis of wood samples collected during surveys in 2002 in Irkutsk, Krasnoyarsk, Sverdlovsk, Novosibirsk and Altai regions revealed only the presence of B. mucronatus. Out of 442 wood samples of pine, larch, fir, B. mucronatus was found 42 times, i.e. in Irkutsk (in 20% of the samples), Krasnoyarsk (1%), Sverdlovsk region (31%), Novosibirsk (8%), and Altai (10%). For the 247 conifer wood samples analyzed from the Krasnoyarsk region B.mucronatus was found only three times. The differentiation of the pinewood nematode B. xylophilus and relative species, B. mucronatus and B. fraudulentus, based only on morphological parameters is difficult or sometimes impossible. The genetic diversity of specimens from twelve isolates from all the mentioned regions maintained on Botrytis cinerea was studied. PCR-RFLPs and sequences of ITS-rDNA confirmed the morphological identification and revealed that all studied isolates belong to the European genotype of B. mucronatus. (RFBR grant No. 01-04-49121).

Evidence from SSU rDNA of Anoplostoma rectospiculum (Galtsova, 1976) suggests that Anoplostomatidae is not related to Oncholaimidae

Anna N. PEGOVA1*, Elena D. KRASNOVA1, Vladimir V. ALEOSHIN2

1Pertsov White Sea Biological Station, Department of Biology, Moscow State University, Moscow, 119899, Russia, *[email protected]

2Belozersky Institute of Physico-Chemical Biology, Department of Evolutionary Biochemistry, Moscow State University, Moscow, 119992, [email protected]

Nematodes from the order Enoplida constitute a mainly free-living marine group of species for which there are no morphological synapomorphies currently available for their higher-rank classification. Thus, the small enoplid family Anoplostomatidae was usually classified within the superfamily Oncholaimoidea, but, alternatively, it was also being placed inside the Enoploidea (Lorenzen, 1994). The family possesses some features unique or rare within Enoplida. These are: a spacious toothless buccal cavity surrounded by pharyngeal tissue in the posterior section only, a cephalic capsule without an insertion point for musculature and a copulatory bursa in males. We analysed the position of Anoplostoma rectospiculum in a dataset comprising all enoplid sequences published so far, which belong to three monophyletic superfamilies (Enoploidea, Oncholaimoidea and Tripyloididea). MP, ML and NJ algorithms with bootstrap replicates consistently infer A. rectospiculum as a member of Enoploidea despite long branch leading to this species on trees. This topology is supported by an unique 1-bp insertion shared by A. rectospiculum and other members of Enoploidea. This character can be used for identification of 18S rRNA sequences of Enoploidea. (RFBR grants No. 01-04-48832, 02-04-48958; S.Sc. grant No. 1712.2003.4)

The SSU rRNA gene as a phylogenetic marker for major groups of Nematoda

Leonid Y. ROUSSINE1*, Paul De LEY2, Mark BLAXTER3, Andy VIERSTRAETE4, Vladimir V. ALEOSHIN1

1Belozersky Institute of Physico-Chemical Biology, Department of Evolutionary Biochemistry, Moscow State University, 119992, Moscow, Russia,*[email protected]

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2Department of Nematology, University of California, Riverside, CA 92521, USA, [email protected]

3Institute of Cell, Animal and Population Biology, University of Edinburgh, Edinburgh EH9 3JT, UK, [email protected]

4University of Ghent, Department of Biology, B-9000 Gent, Belgium, [email protected]

SSU rDNA data has been used in universal phylogenetic studies to define major lineages within Nematoda and suggest a new scenario of their relationships. However, since first attempts to recover deeper nematode phylogenies it has become clear that conventional techniques of inference often are not sufficient for this purpose. Intensive molecular phylogenetic analysis of the three major nematode clades demonstrated that relationships within Chromadoria are robustly inferred down to the deepest divergences. The deeper phylogenetic structure of Enoplia, on the contrary, is not robustly recovered even with the most sophisticated algorithms. The similar situation, albeit to a lesser extent, is characteristic of the Dorylaimia clade represented by only a few compact recent taxa. Our analyses suggest that the behaviour of the SSU rRNA gene is fundamentally similar in all nematode taxa. The main cause of bias in reconstructions of enoplian and dorylaimian phylogenies is a marked scarcity of reliable molecular characters to define deeper nodes of the tree, which is most likely due to poor taxon sampling. The current availability of evolutionary descendants of mile-stone divergences in Chromadoria permits the breaking of long inner branches of the tree and thus avoids computational artifacts caused, in particular, by disparities in rates of molecular evolution. A shortage of recent intermediate taxa in Enoplia appears to preclude similar solutions, and may result from their rapid initial radiation with subsequent extinction of greater number of descendant lineages. A similar evolutionary pattern is probably characteristic of Dorylaimia, although the amount of their recent taxa is very few. The latter allowes one to individually assess few competing phylogenies for reliability. (Grants RFBR No. 01-04-48832, 02-04-48958; RRSCCE No. 1712.2003.4; FEBS Collaborative scholarship for Central & Eastern Europe).

The sequence analysis of D2-D3 domain of LSU rDNA of Thelastomatidae (Thelastomatoidea, Oxyurida) from hind gut of cockroaches

Sergei E. SPIRIDONOV1, Elena A. GUZEYEVA2, Sergei V. LUKYANTSEV2

1Institute of Parasitology of RAS, Leninskii pr., 33, Moscow, 117071, Russia, [email protected]

2Laboratory of Entomology, Department of Plant protection, Tomsk State University, pr. Lenina, 36, Tomsk, 634050, Russia, [email protected]

The sequences for 739-788 bp long fragments of LSU rDNA D2-D3 domain were obtained for 6 species of cockroach thelastomatids (Cranifera cranifera, Hammerschmidtiella diesingi, Hammerschmidtiella sp. from Gromphadorhina portentosae cockroaches, Leidynema appendiculata, L. porentosae, and unidentified Thelastomatidae females from Blaptica dubia), and single Oxyuridae from toads, Thelandros sp. (to use as outgroup). The BLAST search in GenBank revealed sequences for Acrobeloides and Cervidellus (Rhabditida) as the closest ones for studied oxyurids. Maximum parsimony analysis revealed moderate support for the monophyly of Thelastomatidae, but 100% bootstrap support for the monophyly of Oxyuridae and Thelastomatidae. The close relationships of Cranifera and Leidynema were strongly supported, with unidentified Thelastomatidae clustering with Cranifera under weak bootstrap support. Total nucleotide differences in 8-10% (60-75 bp) were reported between species (with 8-16 nucleotide autapomorphies for 5 thelastomatids identified up to the species) and in 15-20% (100-150 bp) between genera. Thelastomatids of the same genus can inhabit hind guts of cockroaches belonging to distant clades of Blattoidea evolutionary trees (Kambhampati, 1995; Grandcolas and D’Haese, 2001). Prominent molecular and morphological differences were shown for Leidynema appendiculata and L. portentosae parasitizing cockroaches from separate evolutionary lines (Blattidae vs. Blaberidae). Also D2-D3 sequence of Hammerschmidtiella specimens from Gromphadorhina portentosae (Blaberidae) was found to be significantly different from that of H. diesingi inhabiting Blatta orientalis and Periplaneta americana (Blattidae). Combined molecular and morphological characters support the independence of Hammerschmidtiella species from G. portentosae.

Molecular and morphological characterization of new EPN isolates from Western Siberia

Sergei E. SPIRIDONOV1, Maurice MOENS 2

1Institute of Parasitology of RAS, Leninskii pr., 33, Moscow, 117071, Russia, [email protected]

2Agricultural Research Center, Burg. Van Gansberghelaan 96, 9820 Merelbeke, Belgium, [email protected]

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Eleven cultures of entomopathogenic soil nematodes (9 of Steinernematidae and 2 of Heterorhabditidae) were established during the field collection work on the territory of Altai Republic (Gorny Altai), Novosibirsk and Tomsk regions in Western Siberia, Russia. Cultures were established through traditional baiting technique and through direct extraction of infective juveniles from soil samples originated from natural habitats ("black taiga" forest on different altitudes, meadows) and agricultural ecosystems. DNA extraction and PCR amplification of ribosomal genes were done to obtain full sequence of ITS1+5.8S+ITS2 fragment of ribosomal genes for 8 steinernematid and two heterorhabditid isolates. Two isolates of S. feltiae were found in Altai Republic and Tomsk Region (both in agricultural habitats). Analysis of sequence data revealed that S. feltiae culture isolated in Tomsk corresponds to the type one from North-Eastern Russia (Izhevsk), when that from Gorny Altai - to the form of this species more common in South-Western Europe and Asia. Two steinernematid species were isolated in natural habitats of Altai mountains: several cultures of S. kraussei and steinernematids morphologically similar to the S. intermedium, but with differences from this latter in ITS rDNA sequence. In their ITS rDNA sequences S. kraussei cultures from Altai mountains are close to conspecific isolates from Switzerland and Scotland and "intermedium-like" cultures - to morphologically similar isolates from UK (E1), Russia (Zvenigorod), Estonia (E6) and Belgium (Rochefort). Both isolated Heterorhabditis cultures (Gorny Altai and Novosibirsk) belong to the H. megidis, what represents first molecularly-confirmed report of this species from Russia. (RFBR grant No. 02-04-48389).

Preliminary results of ITS rDNA sequence analysis for steinernematid strains (Rhabditida; Steinernematidae) from Russian Far East

Sergei E. SPIRIDONOV1, Mutsuhiro YOSHIDA2, Maurice MOENS3

1Institute of Parasitology of RAS, Leninskii pr., 33, Moscow, 117071, Russia, [email protected]

2National Institute for Agro-Environmental Sciences 3-1-3, Kannondai Tsukuba, Ibaraki 305-8604, Japan, [email protected]

3Agricultural Research Center, Burg. Van Gansberghelaan 96, 9820 Merelbeke, Belgium, [email protected]

Full sequences of the ITS1+5.8S+ITS2 domain of rDNA were obtained for the Sakhalin isolate of Steinernema feltiae, four isolates of S. kraussei from Russian Far East (Sakhalin and Primorski Territory) and one S. feltiae isolate from Hokkaido. Sequences for S. kraussei were obtained through direct sequencing of PCR product; those for S. feltiae were obtained by cloning of the PCR product in Escherichia coli competent cells. The comparison of obtained sequences with ITS rDNA sequences for other S. feltiae and S. kraussei isolates was performed through maximum parsimony analysis. All four S. kraussei isolates clustered together under moderate bootstrap support. They form a strongly supported group of isolates together with S. kraussei isolates from Western Siberia and some S. kraussei from Europe. All these Far Eastern S. kraussei isolates were characterised by the presence of an identical unique insertion in ITS2 rDNA. Several characteristic nucleotide substitutions were also found in the ITS1 rDNA of these strains. One such substitution was also reported for S. kraussei from Northern Europe (North East of Russia and Iceland), when two others were only present in two unusual Steinernema populations of the "kraussei" group from Europe (probably new steinernematid species). Both sequenced S. feltiae isolates clustered under strong bootstrap support with the topotype isolate of S. feltiae from North-East of European Russia, although the characteristic 10 bp insertion in the ITS1 rDNA was absent in these Far Eastern isolates. This insertion is reported for several S. feltiae strains common in Asia, Australia but is rarely reported from Europe.

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ENTOMOPATHOGENIC NEMATODES

A representation of Drilonematoidea (Rhabditida) biodiversity in museum collections

Elena S. IVANOVA, Sergei E. SPIRIDONOV*

Institute of Parasitology of RAS, Leninskii pr., 33, Moscow, 117071, Russia, *[email protected]

Four museum collections of earthworms from Moscow State University Zoological Museum, Smithsonian Natural History Museum (Washington, USA), Musйum Histoire d’ Naturelle (Genйve, Switzerland) and Musйe Royal de l’Afrique Centrale (Tervuren, Belgique) were studied for presence of nematodes (Drilonematoidea) parasitizing in body cavity of these invertebrates. Formalin-preserved earthworms were dissected and parasites were recovered. Each earthworm collection except Tervuren one represents the local annelid fauna as well as the animals from around the world. The latter part of collections reflects mainly the scientific interests of researchers contributed. If the bulk of collection in Smithsonian consists from megascolecids from South East Asia, there are South American glossoscolecids and acanthodrilids in Geneva and Indochina megascolecids in Moscow and African octochaetids and eudrilids in Tervuren. Only 3-4% of earthworms in collections was found infected by parasitic Drolonematoidea, whereas infectivity varies in different annelid taxa. About 23% of megascolecid and tropical acanthodrilid earthworms and 2.5% of glossocolecids and 3% of octochaetids and eudrilids and less than 1% of lumbricids were found infected. If megascolecids and acanthodrilids serve as hosts for several dozens genera of Drilonematidae, Ungellidae, Homungellidae and Scolecophilidae families, there are members of only two genera of Drilonematidae parasitic in lumbricids, representatives of few ungellid genera in glossoscolecids and some ungellid and drilonematid genera inhabiting in octochaetids and eudrilids. From few to couple of dozens nematode specimens can be recovered usually from body cavity of earthworms from museum collections though maximum infectivity found in natural habitat was nearly four hundreds nematodes per earthworm.

The "intermedium-affine" group of Steinernema (Rhabditida, Steinernematidae): morphological and molecular characters

Elena S. IVANOVA, Sergei E. SPIRIDONOV*

Institute of Parasitology of RAS, Leninskii pr., 33, Moscow, 117071, Russia, *[email protected]

Steinernematids of "intermedium-affine" group are a separate clade in the genus phylogeny, consisting of two described species and numbers of poorly distinguishable isolates from Holarctic. Morphology of adults and infective juveniles was studied and ITS rDNA sequences were obtained for 4 European isolates of S. affine, 2 isolates of S. intermedium from USA, and 8 unidentified Eurasian isolates. According to the maximum parsimony analysis all Eurasian isolates clustered together, with S. affine forming strongly supported group with Estonian isolate EE3 and Swiss CH221. Type USA isolate of S. intermedium (Poinar, 1985) differs from European "intermedium-like" cultures by shorter male stoma (2.7 μm vs 8.4 μm in cultures close to UK E1 vs 4.5 μm in Caucasus isolate vs 5.5 μm in EE3). All Eurasian isolates were characterized by the presence of mucron on male tail (1-2 μm long in E1 UK and close cultures, 1-5 μm in EE3 and 4-12 μ in Caucasus isolate). The latter one from Caucasus ridge differs from all other studied isolates by having larger and stronger spicules with wider spicule tips and longer tail mucron (4-12 μm). This isolate demonstrates the highest number of nucleotide autapomorphies and total nucleotide differences. Nematodes of the isolates similar to UK E1 presented separate clade in the trees with 100% bootstrap support, with minor total nucleotide differences between separate isolates.

Contortylenchus genitalicola (Tylenchida: Allantonematidae): the sphaerularioid nematode with a fungal feeding phase and the potential biological control agent of the Japanese pine sawyer

Hajime KOSAKA

Department of Forest Microbiology, Forestry and Forest Products Research Institute, Tsukuba 305-8687, Japan, [email protected]

Contortylenchus genitalicola Kosaka & Ogura, 1993 is one of the insect parasitic nematodes of the order Tylenchida (sphaerularioid nematodes). This nematode can develop not only through an insect parasitic phase but also through a fungal feeding phase. The insect host of C. genitalicola is the Japanese pine sawyer, Monochamus alternatus Hope, a serious forest pest and the vector of the pine wood nematode, Bursaphelenchus xylophilus (Steiner & Buhrer) Nickle, 1970, the cause of pine wilt disease, a serious disease worldwide. The fungal host of C. genitalicola is an unidentified fungus that was isolated from the single juvenile nematode recovered from the adult host insect. First in this presentation, the basic biology of C. genitalicola, the morphology and life cycle, is reported. Then, the potentials of C. genitalicola as the

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biological control agent, such as the effects on the host insect, geographical distribution and mass production through the fungal feeding phase, will be presented. The taxonomic status of C. genitalicola would be delicate because different researchers allocate the fungal feeding sphaerularioids to different taxa. There are also some obstacles to develop the biological control using C. genitalicola. Further studies to break these barriers are discussed.

Nematode fauna from hind gut of diplopods from Guadeloupe, French West India

Svetlana V. MALYSHEVA1, Sergei E. SPIRIDONOV2

1Department of Invertebrate Zoology, Faculty of Biology, Moscow State University, Moscow, 119899

2Institute of Parasitology of RAS, Leninskii pr., 33, Moscow, 117071, Russia, [email protected]

Specimens belonging to four species of diplopods were collected in July 1996 in Guadeloupe, French West India. Leptogoniulus naresi (Pocock, 1893) and Trigoniulus lumbricinus (Gerstacker, 1873) were collected from compost heaps behind households in Gosier, Grande Terre. Anadenobolus politus (Porat, 1889) and Pseudospirobollellus avernus (Butler, 1876) were collected under leaf litter in the tropical forest near "Maison de la forкt" in Basse Terre. Twelve dissected L. naresi contained 3 species of nematodes: Heth mauriesi, Rhigonema caribae and Ruizia karukerae. H. mauriesi, Thelastoma sp. and two early juveniles of Rhigonema sp. were found in fifteen specimens of T. lumbricinus collected together with L. naresi. Thelastomatid nematodes of unknown genus were found in 9 dissected specimens of P. avernus, being accompanied with separate Galinanema females, few specimens of Heth and Rhigonematidae juveniles. Six specimens of large A. politus contained the richest fauna of nematodes: Heth mauriesi, Carnoya kermarreci, Ichthyocephalus anadenoboli, Rhigonema sp., Galinanema sp., Thelastoma sp. and unknown thelastomatid. Infection intensity varied significantly in A. politus: when two specimens contained several dozens of Ichthyocephalus anadenoboli males, females and juveniles, about a dozen of adult Heth maurieci and Carnoya kermarreci specimens and numerous juveniles of rhigonematids and thelastomatids, whereas four remaining A. politus contained only few juveniles. Observed differences in nematode fauna in both pairs of sympathric diplopods indicate that some host specificity exists in thelastomatid and rhigonematid nematodes. It seems that some exotic nematodes (Carnoya, Galinanema) can be found only in primary tropical habitats.

Entomoparasitic tylenchids Spilotylenchus (Sphaerularioidea, Tylenchida) cause malformations in male reproductive system of flea Coptopsylla lamellifer (Siphonaptera)

Olga V. SLOBODYANYUK

Institute of Parasitology of RAS, Leninskii pr., 33, Moscow, 117071, Russia, [email protected]

The nematode Spilotylenchus pawlowskyi (Spilotylenchidae, Sphaerularioidea) is parasite of a body cavity of flea Coptopsylla lamellifer which is ectoparasite of gerbils Meriones meridianus and M. tamariscinus. It was shown that the nematode infection caused the malformations of male flea reproductive system and modified segments of the insect hosts. Majority of infected flea males were found sterilized by reduction of phallosome and absense of testes. Also the lack of aedeagus and penis apophyses were often observed as well as the various malformations of modified segments, which represent normally a complex organ for fixing flea female during copulation. Measuring decrease, shape deformation and non-typical telomere armament, basimere narrowing, atypical set of basimere distal setae, manubrium and apodeme malformations were discovered in infected males. Shape deformation, measuring descrease, setae reduction of IX sternite horizontal arm, reduction and non-characteristic shape of IX sternite vertical arm, number and length of antesensill setae decrease, particularly the lowest one, the erratic shape and armament of VIII sternite and occasionally loss of bilateral symmetry were also revealed in infected flea males under teratogenic pressure. Amount of S. pawlowskyi juveniles in host abdomen was found not to affect on nematode teratogeneity though their number differ from few to several thousands juveniles. However some correlation between number of parasitic females penetrated into host haemocoel and number and degree of some malformations were revealed in flea males invaded by four and more parasitic females. Parasitogenous malformations of male reproductive systems caused unability of males to copulate with females.

The influence of Spilotylenchus (Sphaerularioidea, Tylenchida) nematode infection on flea Coptopsylla lamellifer (Siphonaptera, Coptopsyllidae) female reproductive system

Olga V. SLOBODYANYUK

Institute of Parasitology of RAS, Leninskii pr., 33, Moscow, 117071, Russia, [email protected]

The teratogenic effect of Spilotylenchus pawlowskyi (Sphaerularioidea, Spilotylenchidae) on female sexual organs and modified segments of flea Coptopsylla lamellifer was studied. Infected fleas were collected in the

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Astrakhan region, in Aral lake area and Muyunkum desert. Most of examined flea females were found sexually abnormal and lacking ovaries and the single female C. lamellifer from Muyunkum desert was found having the underdeveloped ovaries with no signs of undergoing oogenesis. A set of malformations such as deformation of spermathecs, vagina shortening, reduction and different degree of sclerotization of copulative duct (ductus bursae copulatricis), shape deformation and non-typical armament of anal segments, lesser length and number of antesensill seta were found in reproductive system of infected flea females. Also the bilateral symmetry disturbances of the modified segments were discovered in some flea female specimens. In fact, the teratogenic effect of S pawlowskyi on flea female C. lamellifer sexual organs is as large as causing inability of infected flea females for reproduction. Some taxonomically significant flea features such as number and shape of spermathecs and shape of copulative duct could be so much changed by teratogenic effect that it made difficult the identification of infected fleas.

Teratogenic effect of entomoparasitic tylenchids (Sphaerularioidea, Spilotylenchidae) in three populations of plague-vector fleas Coptopsylla lamellifer (Siphonaptera, Coptopsyllidae)

Olga V. SLOBODYANYUK1, Nadezda L. GERSHKOVICH2

1Institute of Parasitology of RAS, Leninskii pr., 33, Moscow, 117071, Russia, [email protected]

2Zoological Museum of Moscow State University, Bolshaya Nikitskaya str., 6, Moscow,103009, Russia

The nematode Spilotylenchus pawlowskyi (Sphaerularioidea, Tylenchida) is parasite in a body cavity of plague-vector flea Coptopsylla lamellifer which are ectoparasites of gerbils Meriones meridianus and M. tamariscinus. Comparative study of pathomorphology of fleas infected by entomoparasitic tylenchids from three plague natural habitats has been carried out. Infected fleas C. lamellifer dubinini were obtained in Astrakhan region, in Aral lake area and Muyunkum desert ("astrakhan", "aral" and "muyuncum" populations respectively). 36 morphological characters of flea male reproductive system and modified segments and 14 ones of females were studied and it was found that nematodes infection caused strong malformations in all populations studied. However, teratogenic effect in three populations differs from the most impact in "astrakhan" population to the least one in "muyuncum" population. Analysis of morphometry of three insect populations revealed reliable differences between populations based on characters of flea male morphology such as size and shape of phallosome rudiments, as well as manubrium and apodeme of IX tergite and vertical arm of IX sternite. The presence of testes, aedeagus rudiments, and penis apophyses were discovered only in some infected male specimens in "muyuncum" population whereas no males with these organs were found in two other populations. The lack of basimere and intersexual characters were discovered only in some infected male specimens in "aral" population. Female pathomorphology is less of males one in the studied populations. Size and shape of sclerotized copulative duct (ductus bursae copulatricis) and loss of bilateral symmetry were considered as the most affected features.

Comparative study of three coastal steinernematids, Steinernema feltiae, Steinernema sp. RFLP type MY2 and Steinernema sp. RFLP type MY8

Mutsuhiro YOSHIDA

National Institute for Agro-Environmental Sciences, Kannondai Tsukuba, Ibaraki, Japan, [email protected]

As a result of a series of survey for the entomopathogenic nematodes in Japan and Russian Far East, Steinernema feltiae and Steinernema. spp. RFLP types MY2 and MY8 were considered to be coastal steinernematids in the survey regions and many isolates of them were detected from grassland in sandy beach. S. feltiae was isolated from southern part of Sakhalin and Hokkaido, Steinernema sp. MY2 from Hokkaido to central part of Honshu and Steinernema sp. MY8 from central part of Honshu to Amami-Oshima Is., south of Kyushu. S. feltiae and S. sp. MY2 were isolated sympatrically in Hokkaido, while Steinernema sp. MY2 and Steinernema sp. MY8 were isolated allopatrically from the same coast in the boundary area of their distribution. S. feltiae and Steinernema sp. MY2 showed many morphological similarities. From the lateral field pattern of infective juveniles, Steinernema spp. MY2 and MY8 were sorted into a group, which belonged to the glaseri group, but they were clearly discriminated morphologically from each other. As for pathogenicity for some species of lepidopteran larvae, Steinernema sp. MY8 showed much lower mortality as compared with S. feltiae and Steinernema sp. MY2. S. feltiae and Honshu isolates of Steinernema sp. MY2 showed high pathogenicity at 10°C, however Hokkaido isolates of Steinernema sp. MY2 showed low pathogenicity at 10°C. In this report, the biological relationship among two morphological related species and the third species, which showed the same habitat preference as the former two species, will be discussed with some data on their distribution and pathogenicity.

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ECOLOGY OF AQUATIC NEMATODES

Preliminary data on free-living marine nematode distribution in the northern part of the Sea of Japan

Natalia P. FADEEVA, Natalia L. DEMCHENKO

Far East State University, Vladivostok, 690600, Russia, [email protected]

The northern part of the Sea of Japan is the most productive and sensitive to modern negatative anthropogenic activity zone. Qualitative samples of meiobenthos were collected during expedition to Tatarskyi Strait from June to August 2001 from the depth of 12-160 m. Nematofauna abundance ranged from 2 to 1350 individuals per 10 cm2 in Tatarskyi Strait and 278-933 individuals per 10 cm2 in Chichacheva Bay. The taxonomic list of species of free-living marine nematodes of northern part of the Sea of Japan includes about 80 species. Nematodes of the genus Setosabatieria, Metasphaerolaimus, Neotonchoides, Camacolaimus, Leptolaimus, Prochaetosoma and other are recorded for the first time in this region. Nematode communities were identified using classification and ordination techniques. The identified communities were characterized in connection with species composition, density, diversity, size structure and feeding type. Deposit feeders (80%) were most dominant in all communities followed by omnivore/predators, that is explained by the muddy nature of the sediment. Epistrate feeders had the lowest abundance (11%). Contemporary status and anthropogenic impact tendencies in the coastal nematofauna are described. Main anthropogenic factors impacting the habitats of these animals are determined.

Biodiversity of Free Living Marine Nematodes From Tropical Brazilian Ecosystems

Verфnica da FONSКCA- GENEVOIS1, Nic SMOL2, Virбg VENEKEY1, Alessandra Prates BOTELHO1

1Departmento de Zoologia, Universidade Federal de Pernambuco, Av. Prof. Moraes Rкgo s/n, Cidade Universitбria, 50000-000, Recife-PE, Brasil, [email protected]

2PINC, University of Gent, Ledeganckstraat, 35; B-9000, Gent, Belgium, [email protected]

Free living aquatic nematodes from Brazil were described for the first time in 1950s by Gerlach (1954; 1956a, b; 1957a, b). These papers represented an important taxonomic contribution as 113 new species were described. Only since the 90s the free-living aquatic nematodes from Brazil were again studied by Brazilian researchers from different institutes, such as Medeiros (1997), Moellmann (2003), Pinto (2003) at the South and Southeast areas of Brazil. In this work a great variety of tropical ecosystems, most of them from North and Northeast of Brazil were considered, such as sandy beaches (Bezerra, 2000), salt ponds (Silva, 2001), Roccas atoll (Netto, 1999), estuaries (Rodrigues, 2002; Castro, 2003), tidal (Esteves, 2002), beach-rocks (Maranhгo, 2003), upwelling region (Fonsкca-Genevois, 2001; Santos, 2002; Nunes, 2003) and algae banks (Da Rocha, 2003). Considering nematodes spatial distribution to-day a list of 203 nematode genera were registered, within them 125 are new records to Brazil. Only the genera Daptonema, Spirinia and Theristus were common to all the ecosystems. The highest number of nematode genera was found at the unique upwelling region of the Brazilian coast.

Marine nematode assemblages and zonation patterns on European (un)disturbed sandy beaches

Tom GHESKIERE, Steven DEGRAER, Magda VINCX

Marine Biology Section, Biology Department, Ghent University, Gent, Belgium, [email protected]

The European coastline consists for more than 30 % of sandy beaches. In spite of their rather barren and desert-like view, these sandy coastlines harbour a highly diverse fauna and flora. On the other hand these beaches are subjected to strong anthropogenic pressure (e.g. pollution, eutrophication, coastal fisheries and tourism), which has substantial impact on the interstitial life and functioning of the sandy beach ecosystem. In contrast to the well-documented tropical sandy beaches, little is known about the structural and functional diversity of the different benthic components. This study aims to get an idea about the structural and functional diversity of the free-living marine nematodes of three European sandy beaches (i.e. Belgium, Poland and Italy). Nematodes are very suitable for monitoring and are used in the second part of this study to compare and evaluate the diversity between "disturbed" and "undisturbed" sandy beaches. In order to document the structural and functional diversity of the nematofauna of the above-mentioned beaches, quantitative samples along transects have been collected on both disturbed and undisturbed parts of the beaches. Nematodes species were processed and identified at species level and further analysed by means of statistical and multivariate techniques. Around 250 species of free-living marine nematodes in total have been recognized on these beaches of which several species are new to science. Nematode zonation patterns, in close relation with the morphodynamics of the beaches, have been detected as well.

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The seasonal changes of free-living nematodes from the deep part of the White Sea

Maria A. MILJUTINA

Department of Biology, Moscow State University, Moscow 119899, Russia, [email protected]

Seasonal changes in quantity, diversity and distribution of free-living nematodes were described for deep part of the White Sea at the depth of 270 m. Nematodes were collected in July and October, 1998, in May and November, 1999. Nematodes were the most abundant group among other metazoans (about 70% of all metazoans). The rate of nematodes in spring and summer was about 65%, while in autumn and winter - 74%. The maximum density of nematodes was found in October (825000 specimens per m2 within 5-cm columns of sediment). Vertical distribution in sediments varied in seasons too. In May and July the most of nematodes concentrated in upper fist centimeter of sediment, in October and November - in the second centimeter. Four species of nematodes: Sabatieria ornata, Filipjeva filipjevi, Microlaimus sp. and Sphaerolaimus gracilis, were the most numerous in all samples. The dominating species differed from season to season because of great seasonal changes in Sabatieria ornata abundance (the number of specimens per 1 m2 in July was 160191, in October - only 19108). Abundance of other numerous species changed less. However a peak of quantity of Microlaimus sp. and Sphaerolaimus gracilis (in July) occurred too. Filipjeva filipjevi had two peaks of high abundance - in July and in November. (INTAS grant No. 96-1359).

Nematode community structure in Vrangel Bay (Nakhodka Bay, Sea of Japan)

Olga N. PAVLYUK1, Yulia A. TREBUKHOVA1, Vladimir M. SHULKIN2

1Institute of Marine Biology FEB RAS, Vladivostok, 690041, Russia, [email protected]

2Pacific Institute of Geography FEB RAS, Vladivostok, 690041, Russia

One of the largest deep-water ports in Russia is situated in Vrangel Bay. Since 1973 till the present time the bottom dredgings are being carried out there. In 1989 and 1995 the structural changes in the soft bottom communities were studied with the exception of meiofauna. In 2001 there was conducted a study of meiobenthos and its main group - free-living marine nematodes. The main aim of this research was to investigate the species composition and quantitative distribution of nematode community structure according to the grain size composition and sediment concentrations of heavy metals. The soft bottom meiofauna in Vrangel Bay was dominated by nematodes (50-80%). Density of nematode population was higher in muddy sediments than in muddy sands. On the whole, 48 nematode species were found. Dorylaimopsis peculiaris, Viscosia stenostoma, Axonolaimus seticaudatus, Metasphaerolaimus japonicus and Pseudosteineria inaequispiculata were dominated in sandy sediments, while muddy sediments were dominated by Sabatieria pulchra, S. palmaris, Metalaimus pumilus, Sphaerolaimus limosus and Oncholaimium ramosum. There was found an insignificant correlation between the abundance of two species (S. pulchra, S. palmaris), grain size composition and concentrations of lead. However, metal concentrations determining in Vrangel Bay were not high enough to influence on species composition and community structure of nematodes.

Nematode communities from the Vostok Bay (Sea of Japan)

Olga N. PAVLYUK, Yulia A. TREBUKHOVA

Institute of Marine Biology FEB RAS, Vladivostok, 690041, Russia, [email protected]

Nematode communities were studied in relation to the sediment granulometry in Vostok Bay (Peter the Great Bay). There were found 8 types of the bottom sediments. In general, nematode diversity and total abundance were regulated by sediment granulometry. The greatest nematode population density was found in fine-grained sand (130 thousands specimens/m2). Extremely low abundance was recorded in fine gravel with an admixture of mixed-grained sand (10 thousands specimens/m2). In total, 89 nematode species were found at all stations in Vostok Bay. The species diversity indices were the highest in silted medium fine-grained sand (H=4.9; e=2.5). In this biotope we registered 64 nematode species totally. The following species were dominated there: Metachromadora itoi, Dorylaimopsis peculiaris, Viscosia stenostoma, Phanoderma platonovae and Axonolaimus seticaudatus. The lowest species diversity indices (H=1.49; e=0.76) were recorded in fine gravel with an admixture of mixed-grained sand, dominated by Dorylaimopsis peculiaris and Axonolaimus seticaudatus. There were found only 4 species. One should notes the structure of nematode communities from Gaydamak Bay, which is situated in Vostok Bay. The bottom sediments are represented there by heterogeneous silt and characterized by high concentrations of heavy metals and petroleum hydrocarbons. A number of the nematode species was impoverished and submitted by the one species, Oncholaimium ramosum. O. ramosum can tolerate the whole range of organic contaminants, which are not toxic and may be used for nutrition. At the

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exit from the bay the composition of the species was more varied. Thus, changes in nematode communities were correlated with the sediment structure and, in the case with Gaydamak Bay, with high concentrations of heavy metals and petroleum hydrocarbons.

A comparison of the nematode communities associated with cold-water and tropical coral rubble

Maarten RAES1, Johan KEMPS2, Ann VANREUSEL3

Marine Biology Section, Ghent University, 9000 Gent, Belgium, [email protected], [email protected], [email protected]

The great variety of species in local areas of coral reefs is legendary, although huge differences in local and regional biodiversity do occur. Recently, much attention is going to the description of the composition and dynamics of faunal communities associated with cold-water corals (mainly Lophelia pertusa (Linnaeus, 1758)). Logically, these coral communities should be subject to analogous processes that result in high habitat complexity (and therefore higher biodiversity) in tropical coral reefs. It was already suggested for macro- and megafauna that cold-water corals rival their tropical counterparts in terms of species richness and diversity. For the first time ever, attention goes to the meiofauna associated with cold-water coral rubble and associated substrates (sponge skeletons of Aphrocallistes bocagei Schultze, 1886) from the Porcupine Seabight (North Atlantic). The nematode community is discussed in terms of biodiversity, community structure and preferences for the different substrate types. This information is compared with that of the community associated with tropical coral rubble from the South Kenya coast, an area with very different environmental conditions. Despite of these enormous differences, some parallels between the 2 communities could be drawn. In this context, the role of the 2 closely related families of epifaunal nematodes Epsilonematidae and Draconematidae is striking: although seldom found in deep-sea environments, these two families were represented by a large number of individuals on cold-water coral rubble from a depth of 1005 m.

Nematode population in a detached kelp accumulation in the White Sea

Alexey V. TCHESUNOV1, K.A. FILIPPOVA1, Vadim O. MOKIEVSKY2

1Department of Invertebrate Zoology, Faculty of Biology, Moscow State University, Moscow, Russia, [email protected]

2P. P. Shirshov Institute of Oceanology, Russian Academy of Sciences, Moscow, Russia

Nematodes were studied in a long-term accumulation of detached kelp situated at depth 18 m near Karelian Coast. The stack consists of fronds of brown algae Laminaria and Alaria originating from a belt of growing plants hereabouts. The stack was tens meters both long and wide and 2 m high. Upper layer of the stack was well aerated and formed by fresh and even living fronds. Middle layer was anoxic and made up by small pieces of thallii covered by cyanobacteria. Bottom layer was anoxic black semi-liquid organic material. There were totally 40 nematode species found in the accumulation. Living detached algae support nematode community with dominant species normally inhabitating alive macrophytes in the sublittoral zone of the White Sea. Nematode community in dead algae at various stages of decomposition is characterized by strong dominance of bacterivorous Geomonhystera sp. and low percentages of other phytal and benthic species. Geomonhystera sp. is related to common G. disjuncta but differs by much larger size and viviparity. Generally, the nematode diversity decreases and the total nematode abundance increases along the stages of plant decomposition. (RFBR grant No. 03-04-49152).

Nematode population in the coastal seasonal ice of the White Sea

Alexey V. TCHESUNOV*, Darja A. PORTNOVA

1Department of Invertebrate Zoology, Faculty of Biology, Moscow State University, Moscow, Russia, *[email protected]

Abundant nematode populations have been registered in the coastal seasonal ice in Kandalaksha Bay of the White Sea. Ice as a continuous covering appears to the end of November and lasts about six months growing up to 1-1.5 m thick. Ice disappears entirely in the beginning of June. First nematodes were observed in December or in January. Total abundance of nematodes increases gradually from January to May. Almost all specimens were always confined to the lower layer of the ice blocks. This nematode population is significantly more diverse than one in drifting ices of the open Arctic Ocean. The community includes Theristus melnikovi and Cryonema crassum, both known as specific inhabitants of the drifting Ocean ice. Both species are represented by all juvenile and adult stages. C. crassum is documented to have egg clutches with developing embryos within ice. A significant portion of the nematode community is made up by other three or four species of the

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family Monhysteridae. The latter are not yet identified because they are represented only by juvenile specimens. With melting ice, the ice-dwelling nematodes disappeared. Theristus melnikovi, Cryonema crassum and other ice-dwelling Monhysteridae were found in summer and autumn seasons neither in bottom sediments nor in fouling in that area. (RFBR grant No. 03-04-49152).

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VERTEBRATE PARASITIC NEMATODES

Ne matode fauna of small mammals of Kare lia

Valentina S. ANIKANOVA*, Sergey V. BUGMYRIN

Institute of Biology KRC RAS, Petrozavodsk, 185610, Russia, *[email protected]

The faunistic complex of the shrew and rodent nematodes of Karelia includes 16 species, belonging to 6 families: Capillariidae (Capillaria incrassata, C. kutori, C. muris-sylvatici, Eucoleus oesophagicala, Hepaticola soricicola), Strongyloididae (Parastrongyloides winchesi), Heligmosomatidae (Heligmosomum mixtum, Heligmosomoides glareoli, Longistriata codrus, L. didas, L. depressa, L. trus), Soboliphymidae (Soboliphyme soricis), Syphaciidae (Syphacia petrusewiczi, S. vandenbrueli), Anisakidae (Porrocaecum depressum). Nematodes of fam. Heligmosomatidae (6 species) and fam. Capillariidae (5) dominated among observed species. Palearctic species accounted for a half (50%) of total species numbers, cosmopolitans - 31%, holarctic species - 19%. Roundworms of shrews were characterized by the highest diversity of species composition (11), life cycles and localization in a host body. The parasites with a simple cycle of development, living in intestine, prevailed (Anikanova et al., 2001). The development of other nematode species is realized with participation of intermediate, reservoir hosts and disseminators. These nematodes are localized in various organs of shrews - oesophagus, stomach, liver, urinary bladder, and have low level of host infection. Five species of nematodes, having a simple cycle of development and living in intestine, parasitize at rodents (Bugmyrin et al., 2001). The various food specializations of shrews and rodents determine species composition of the nematodes and quantitative indices of host invasion level. The highest species diversity of the nematode complex was revealed at dominating host species, Sorex araneus and Clethrionomys glareolus.

Distribution of nematode Cystidicoloides tennuissima in the fish biocoenoses of Paanajдrvi - Olanga lake-river system

Juliya BARSKAYA, Olga NOVOKHATSKAYA

Karelian Research Center of RAS, Petrozavodsk, Russia, [email protected]

Nematoda Cystidicoloides tennuissima is the typical parasite of Salmonidae fish. Cystidicoloides tennuissima is the parasite of alimentary tract of fish. The parasite fauna of brown trout, white fish and grayling (145 samples) was studied by the common parasitological methods. Analysis of Salmonidae fish parasite fauna showed that the core of the parasite fauna formed in parr 3+. Cystidicoloides tennuissima is the permanent component of the core. The largest abundance index was recorded in parasite fauna of grayling. Cystidicoloides tennuissima is the dominant species in the parasite fauna of this host. It is peculiar that Cystidicoloides tennuissima is registered in the parasite fauna of brown trout or white fish only in the case if waterbodies are inhabited by grayling. The increasing of specific composition of parasite fauna of Salmonidae of older age groups leads to the increasing of number of species in the core. In addition, the species forming the parasite fauna of parr as Cystidicoloides tennuissima remain in the core of parasite fauna of adult specimens. The infection index of grayling by Cystidicoloides tennuissima is still higer. How it was reported when was discussed the fauna of parr, the grayling has the high infection by Cystidicoloides tennuissima. So, the grayling is the main definitive host of Cystidicoloides tennuissima. This host supports the size of population of Cystidicoloides tennuissima.

Nematodes of fishes from freshwater reservoirs of Primorye (Russia)

Alexey V. ERMOLENKO*, Marina B. SHED’KO

Institute of Biology and Soil Sciences FEB RAS, Vladivostok, 690022, Russia, *[email protected]

Forty species of parasitic nematodes were found in the fishes from freshwater reservoirs of Primorye 2400 specimens of 70 fish species were studied). Six species of nematodes are of marine origin, 33 species are of freshwater origin and Oswaldocruzia filiformis had got into the fish by accident. Fish are the intermediate hosts for Gnathostoma spinigerum, Dioctophyme renale, Porrocaecum sp., Cotracaecum sp., Anisakis simplex, Pseudoterranova decipiens. For Raphidascaris acus and Hysterothylacium aduncum fish are the intermediate and definitive hosts. For 32 species of the worms fish are the definitive hosts. In the last case Capillaria salvelini and Cucullanus truttae can be developed without the intermediate hosts, and the life cycle of other species passes through intermediate hosts both planktonic invertebrate (Copepoda, Euphausiacea) and benthonic one (Amphipoda, larvae of the amphibiotic Insecta, Oligochaeta). The highest species diversity of nematodes (29 species) was registered in the fish of Khanka Lake basin. 25 species of the worms (2 species of marine origin) were met in the Ussuri River basin, 20 (4) species were met in the rivers of the southern Primorye, 13 (4) species were met in the rivers of the eastern slope of Sikhote-Alin. The nematodes that were found in the fish from the last area are of holarctic origin only (they belong to the boreal foothill complex,

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boreal flat, pacific foothill and arctic freshwater faunistic complex). On the contrary in the Ussuri River basin and especially in the Khanka Lake basin the representative of the southern species complex (Chinese flat and Indian flat complexes) predominate. In the southern Primorye the number of holarctic species and Chinese- Indian species was approximately equal. Such distribution of the parasites is connected partly with a water body hydrology, but basically - with the historical reasons which have defined an opportunity or impossibility of setting in those or these reservoirs for nematode species of different origin.

Distribution of the swim bladder nematodes of salmonid fishes in the Far East of Russia

Marina B. SHED’KO*, Alexey V. ERMOLENKO

Institute of Biology and Soil Sciences FEB RAS, Vladivostok 690022, Russia, *[email protected]

Up to the present time four species of nematodes of genus Cystidicola and Salvelinema have been recorded from the swim bladder salmonid fish: C. farionis, C. stigmatura, S. salmonicola, S. walkeri.The systematics of these worms has been unusually confusing before their revision. All four species of nematodes are known in watersheds of eastern part of Pacific basin. In the rivers of Japan only S. salmonicola was registered. Cystidicola farionis and Salvelinema salmonicola are undoubtedly marked in various regions of Far East of Russia on the literary and our original data. The samples of 8 genera of salmonid fish were studied in the reservoirs of Primorye, Kamchatka Peninsula, Kurile Islands [Kunashir, Paramushir], South Sakhalin, the Sea of Okhotsk; also the collection of Mamaev and Paruchin (1958) was examined. The area of first of them coincides with distribution of the intermediate host, Pontoporea affinis (Amphipoda, Crustacea), and includes the rivers of Chukchi Peninsula, Kamchatka Peninsula (especially in its eastern part), Amur River basin (main river-bed and northern inflows), the Sea of Okhotsk and probably the water of North Sakhalin. C. farionis was found in various species of genera Salvelinus and Oncorhynchus, in Hypomesus olidus, Parasalmo mykiss, Thymalus mertensi, Osmerus mordax dentex, Coregonus ussuriensis. S. salmonicola is distributed in the southern region of the Kamchatka Peninsula, Kurile Islands, South Sakhalin, in the main river-bed and southern inflows of Amur river (Ussuri river basin) and in Primorye. Gammarus koreanus was registered as the intermediate host of this nematode species. S. salmonicola basically was met in fish of the genera Salvelinus and Oncorhynchus, Brachymystax lenok, Hucho taimen, Thymallus spp., Coregonus spp., and very seldom in Brachymystax tumensis. The peculiarities of infection by nematodes of different hosts are discussed also in the present work.

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TAXONOMY AND SYSTEMATICS

Fauna of marine nematodes from the intertidal of Putyatin Island (Sea of Japan, Peter the Great Bay)

Ludmila S. BELOGUROVA

Institute of Marine Biology FEB RAS, Vladivostok, 690041, Russia

Twenty-two species of free-living nematodes belonging to 18 genera, 10 families and 5 orders, have been found in the intertidal zone of Putyatin Island (Sea of Japan, Peter the Great Bay). The order Enoplida is represented by 14 species, the order Monhysterida by 3 species; the orders Chromadorida and Areolaimida by two species, and the order Desmodorida by one species. The genus Eurystomina is represented by 3 species; the two genera, Adoncholaimus and Axonolaimus, by two species. Only one species was recorded for each of the genera: Phanoderma, Enoplus, Anticoma, Pontonema, Viscosia, Oncholaimium, Oncholaimus, Pseudoncholaimus, Calyptronema, Sabatieria, Paracanthonchus, Monoposthia, Pseudosteineria, Mesotheristus, Terschellingia. Twenty one species of nematodes were found in the lower intertidal horizon in the belt-forming communities of Phyllospadix iwatensis, Sargassum miyabei, Corallina pululifera, that is 95.4% of the total number of nematodes. Nine species were found in muddy sand within the belt-forming communities of Phyllospadix iwatensis. They are: Anticoma behringiana, Adoncholaimus fuscus, Viscosia stenostoma, Eurystomina ophtalmophora, Sabatieria palmaris, Pseudosteineria inaequispiculata, Mesotheristus paracircumscriptus, Terschellingia glabricutis, and Axonolaimus orus. Twelve species were recorded in the middle intertidal horizon in the belt–forming communities of Chaetomorpha moniligera, Enteromorpha clathrata, Scytociphon lomentaria. The nematode fauna of this horizon is represented by the species, which are also found in the lower intertidal horizon, with the exception of one species, Oncholaimium japonicum, recorded only in the middle intertidal horizon in the belt-forming Chaetomorpha moniligera. Four species of nematodes (Enoplus anisospiculis, Pontonema papilliferum, Oncholaimus brachycercus, Monoposthia costata) were found in the high intertidal horizon. These species inhabited both the middle and the lower intertidal horizons.

Estimation of taxonomic biodiversity of freeliving nematodes in the Russian Arctic Seas

Valentina V.GALTSOVA, L.V. KULANGIEVA

Zoological Institute Russian Academy of Sciences, St.Petersburg, [email protected]

Studies on biodiversity estimation frequently encounter the difficulties connected with the differences to the degree in the completeness of the mastery of fauna. The methods that make it possible at least approximately to estimate the degree of the completeness of the taxonomic mastery of concrete regions practically do not exist. There is a common property of the regular distribution of any units or their characteristics under the conditions of the limited space: their sequences form straight line with logarithmic scale of both axes. The concrete region which biodiversity of fauna is in prospect to estimate is such limited space. The numerical ratio of taxa of different ranks also forms the sequence which approaches logarithmic. If we build the graph on the Y-axis of which the logarithms of the number of taxa of this rank are arranged and on the X-axis - order, family, genus, species arranged through the logarithmically diminishing intervals then this graph for the concrete region of the World Ocean will be represented by straight line when the group is sufficiently well studied systematically. At the same time the groups weakly studied systematically do not give this rectilinear graph. Thus the possibility to build graph according to the known number of high taxa and according to this graph appears to estimate the number of taxa of genus and species ranks. Then, after comparing the obtained values with the known facts it is easy to estimate the degree of the completeness of mastery. We have accomplished a similar work for several groups of invertebrates from the Arctic seas of Russia. Most fully studied groups proved to be bivalve molluscs (70%), by least studied - nematodes (28, 5%), whose list at the present time includes 376 species. According to our estimation the assumed number of the nematode species in this region is not less than 1300, i.e. the presence of 1000 extra species of nematodes should be expected.

Nematodes of the family Cyatholaimidae from the Russian coast of the Sea of Japan

Olga I. DASHCHENKO

Far-East State University, Vladivostok, 690600, Russia, [email protected]

The family Cyatholaimidae consists of four subfamilies, which are well distinguished by the cuticle ornamentation and by the shape of the male supplements. Five cyatholaimid species have been registered in the Peter the Great Bay of the Sea of Japan. These nematodes inhabit the silty sand, and only Acanthonchus tridentatus has been found in the biofoulings of concrete plates. Three species: Acanthonchus tridentatus Kito, 1976; Paracanthonchus macrodon Ditlevsen, 1918; Paracyatholaimus pugettensis Wieser and Hopper, 1967, belong to the subfamily Paracanthonchinae. The males of these species have the tubiform supplements. The new

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species, Marilynia maritima sp.n. (subfam. Cyatholaiminae), has five weakly cuticularized goblet-shaped supplements (not always clear). The supplement has intermediate shape between tubiform and goblet-shaped. The second new species, Pomponema gracilis sp.n. (subfam. Pomponematinae), has 15-19 sucker-shaped supplements. The cuticle forms large wrinkles between supplements.

Four species of marine nematodes of the order Desmodorida from the Far Eastern sea

Vladimir V. MORDUCHOVICH*, Natalia P. FADEEVA**

Far East State University, Vladivostok, 690600, Russia, *[email protected], **[email protected]

Marine nematodes of the order Desmodorida are rare and poorly studied group. Four species of desmodorids were described from the Sea of Japan and the Okhotsk Sea. Paramicrolaimus spirulifer Wieser, 1959 is recorded in the Far East seas for the first time. In general aspects (cephalic setae of second and third circle are slender, asymmetrical stoma, oval amphids, number of supplements) males and females agree with the descriptions of this species by Wieser (1959) and Jensen (1978). Female reproductive system is described. Aponema minuta sp. n. is the smallest Aponema species known with a body length less than 500 μm, bent spicules 18 μm around the arc, 20-21 μm from tip to tip, and gubernaculum with sclerotized dorsocaudally directed apophyses. Microlaimus adrianovi sp.n. can also be easily distinquished from all known congeneric species in having transverse slit-like vulva with strong sclerotized margins. Molgolaimus orientalis sp.n. differs from all other molgolaimids by the shape and size of spicules and specific rose-coloured inclusions in the cuticle. The photomicrographs of the species of nematodes by light microsopy are presented with detailed morphological descriptions.

Fauna of free living nematodes from some water bodies in the North of Vietnam

Vu Thanh NGUYEN1, Vladimir N. GAGARIN2

1Department of Nematology, Institute of Ecology and Biological Resources, Vietnam National Center for Natural Sciences and Technology (NCST), Hanoi, Vietnam

2Institute of Inland Waters Biology, Russian Academy of Sciences, Borok, Yaroslavl Prov., Russia

Long time the fauna of free-living aquatic nematodes, including fauna of freshwater and brackish water nematodes and marine nematodes of Vietnam has not been studied. At present, the intensity of investigation of fauna of free-living nematodes in Vietnam water bodies is strongly improved, in connection with creating of the new database for the biological monitoring assessment of water quality from surface watercourse of all river watersheds and the wetland ecosystems of Vietnam. In 2000-2002 nematode fauna from 4 water ecosystems of Nhue River, To Lich River, Cau River and West Lake in Northern Vietnam has been observed and studied. These three rivers are situated not far from Hanoi territory. The Cau River flows through Bac Giang province, Thai Nguyen province, Bac Kan province. Fourteen samples were collected from Nhue River and fourteen from To Lich River, fifteen samples were collected from West Lake and twenty eight samples were also collected from Cau River. All nematode samples were taken at the distance of 1-3 m from the bank (shore) at the depth of 1-2 m with ground-slit sediment. The samples were taken by boat by Ponar grabs (core sampler). Nematode samples were fixed by hot TAF solution, and processed to pure glycerin by a slow evaporation Seinhorst’s method and mounted on permanent glass slides. All 89 species belonging to ten orders were found in the samples from four water bodies, 10 species were described and identified as new for science. Most of the species (29) belong to the order Tylenchida, 19 species belong to the order Dorylaimida. All species could be divided into three ecological groups: (i) fresh-water forms, (ii) saline-water forms and (iii) terrestrial forms. The latter group predominates and is the most numerous - 59 species. The scantiest group is saline-water form - 7 species. 23 another nematode species are fresh-water forms.

The Euchromadora–Parapinnanema complex

Nic SMOL1*, Verфnica da FONSКCA- GENEVOIS2, Tвnia Campinas Bezerra1**, Alessandra Prates BOTELHO4, Maria A. FRANCO3

1PINC, Vakgroep Biologie, Universiteit Gent, Gent, Belgium, [email protected], [email protected]

2Departmento de Zoologia, Universidade Federal de Pernambuco, Cidade Universitбria, Recife-PE, Brasil, [email protected]

3Mariene Biologie, Vakgroep Biologie, Gent, Belgium, [email protected]

The discovery of new Euchromadora–Parapinnanema–like species from Brazil posed problems of generic

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placement. Careful examination of the species of both genera resulted in re-evaluation of the generic characters. The presence of a precloacal raised cuticle in the males can no longer be considered as a distinguishing character between Euchromadora and Parapinnanema, as it occurs in both genera. The degree of sclerotisation of the onchial cavity and the structure of the female reproductive system remain valid characters in differentiating of both genera: in Euchromadora rows of denticles are present in the onchial cavity whereas in Parapinnanema they are absent. Females of Parapinnanema are characterized by having a longitudinal vulva and a strong double sphincter around the uterine chamber, whereas Euchromadora has a transverse vulva and no double sphincter.

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