Newsletter of the Russian Society of Nematologists Abstracts of the Tenth International Symposium of the Russian Society of Nematologists (Russia, Golytsino – Bolshie Vyazemy, 1-5 July, 2013) Текст научной статьи по специальности «Биологические науки»

Russian Journal of Nematology, 2013, 21 (2), 141-164

Newsletter of the Russian Society of Nematologists Abstracts of the Tenth International Symposium of

the Russian Society of Nematologists (Russia, Golytsino - Bolshie Vyazemy, 1-5 July, 2013)

ABD-ELGAWAD M.M.M.1, OMER E.A.2 & ISMAIL R.F.2 Effect of plant extracts of some medicinal plants on phytonematodes in Egypt.

Phyto-extracts have advantages over synthetic nematicides because they contain compounds, usually at lower concentrations than synthetic chemicals, that nematodes are not able to inactivate, and which have multiple modes of action and come from renewable sources. Therefore, ethanol extract of Tagetes lucida, a medicinal plant recently introduced into Egypt, was tested and compared with other plant species Achillea millefolium, Cymbopogon citratus, Artemisia annua and Calendula officinalis for their nematicidal activity against plant-parasitic nematodes of common occurrence. All extracts significantly inhibited motility of Meloidogyne incognita, Criconemella spp., Helicotylenchus spp. and Pratylenchus spp. Achillea annua was generally more effective in reducing the numbers of active nematodes tested than others except T. lucida-root extract, which was superior to herbal extract. When transferred to water, the total percentages of nematodes that regained activity ranged from 15% for T. lucida to 38% for A. millefolium after 24 h and from 4% for C. officinalis to 39% for A. millefolium after 72 h. The extracts inhibited hatching of M. incognita in the range 46.8% for A. millefolium to 88.8% for C. citratus compared to 5.1% at the controls. The nematicidal activity of the isolated materials from the plants was concentration dependent. Further studies are warranted to obtain insights on rates and timing of the possible application as bio-nematicides, as well as the growth parameters of treated plants under greenhouse and then field conditions in Egypt. - 1Plant Pathology Department, National Research Center, El-Tahrir St., Dokki 12622, Giza, Egypt. E-mail: [email protected]; Pharmaceutical Sciences Department, National Research Center, El-Tahrir St., Dokki 12622, Giza, Egypt.

ABD-ELGAWAD M.M.M.1, JIAN H.2, QIAO Y.2 & HAMMAM M.M.A.1 Entomopathogenic nematodes and their potential against insects and phytonematodes in Egypt.

Entomopathogenic nematodes (EPN) can be used in IPM programmes to avoid the hazards of chemical pesticides. Surveys for indigenous EPN resulted in isolating numerous species/strains from different parts of Egypt: Steinernema abbasi, S. carpocapsae, S. kushidai, S. glaseri, Heterorhabditis bacteriofora, H. indica, H. taysearae. This latter species originated from Egypt as a new species and had several geographical isolates in the present study. Target insect pests for EPN in Egypt included Artogia (pieris) rapae, Hellula undalis, Temnorhynchus baal, Sesamia cretica, Plutella xylostella, Rhynchophorus ferrugineus, Spodoptera littoralis, Agrotis ipsilon, Tropinota squalida, Galleria mellonella, and Zeuzera pyrina. Moreover, a two-fold goal, i.e., controlling plant-parasitic nematodes and insect pests simultaneously by EPN, is being tried in Egypt. Possible mechanisms of plant-parasitic nematode suppression by EPN were discussed to obtain optimal application tactics and maximise their field effectiveness. - 1Plant Pathology Department, National Research Center, El-Tahrir St., Dokki 12622, Giza, Egypt. E-mail: [email protected]; 2Plant Pathology Department, China agricultural university, Beijing 100193, China.

ANIKIEVA L. Structure and functioning of the parasitic system 'Toxascaris leonina - mammal host'.

Peculiarities of functioning of the parasitic system nematodes Toxascaris leonina were analysed on the basis of original material. The role of the different age groups of the host animals in maintaining of the number and life cycle of this species under changing conditions were studied. - Institute of biology, Karelian research centre, RAS, Petrozavodsk, 185910, Russia. E-mail: [email protected].

BAICHEVA O.1, UDALOVA ZH.2, ZINOVIEVA S.2, GERASIMOVA N.3 & DAMIANOVA A.1 The effect of compounds of chitosan with salicylic acid on the infection of tomato plants by Meloidogyne incognita.

One of the ways to protect plants against pathogens is the induction of resistance by elicitors and signalling molecules. The tomato Solanum lycopersicum - root-knot nematode Meloidogyne incognita system was employed to study the effect of chitosan, a signalling molecule salicylic acid (SA) and the compound of chitosan with fragments of

SA. It was shown that the investigated compounds of chitosan inhibit development of nematodes in treated plants. When comparing the effect of chitosan derivative, a marked elicitor activity of chitosan with SA and modified chitosan was observed. It is suggested that modification of chemical and spatial structure of the oligomer chain of two pieces of 2-hydroxy-3-methoxybenzyl and pyridoxal, promoted expression of protective genes. - institute for Nuclear Research and Nuclear Energy, Sofia, Bulgaria. E-mail: [email protected]; 2Centre of Parasitology, IPEE RAS; 3Bach Institute of Biochemistry, RAS, Russia.

BELOGUROVA L.S. Meiobenthos of the deep area of Peter the Great bay (Sea of Japan) and communities of free-living nematodes.

A study of the deep-sea meiobenthic communities of Peter the Great Bay of the Sea of Japan was conducted in August 2010 for the first time. Samples were collected at 14 sampling points in the bathyal zone at depths of 270, 400, 600 and 800 m. All samples of bottom sediments contained aleurite-pelite fractions, and numerous dead valves of the diatom algae and radiolarian valves. The meiobenthos was represented by seven taxonomical groups and pseudomeiobenthos by four. Nematodes occurred at all depths. The proportion of nematodes varied from 11.6 to 91.7%, the average being 21% of the meiobenthos total population density. The taxonomical composition of the fauna varied with depth. All trophic groups were present in the area of study: selective detritophages (16%), nonselective dethritophages (26%), scrappers (26%) and predators (32%). In total, 19 species of the free-living marine nematodes, belonging to 18 genera, 11 families, and 4 orders were revealed. The highest species number (12) was found at 400 m depth; the lowest species number (2) at depths of 270 and 600 m. Dorylaimopsis peculiaris was a dominant species. -A.V. Zhirmunsky Institute of Marine Biology, Far Eastern Branch of the Russian Academy of Sciences, Vladivostok, 690059, Russia.

BERT W.1, JANSSEN T.1, SLOS D.1, KOLOMBIA A.Y.1, CLAEYS M.1, COUVREUR M.1, MERMANS C.1, ENSAFI P.1, QUISADO J.S.1, XUE Q.1, KARSSEN G.2, YUSHIN V.V.3 & DECRAEMER W.1, 4. Nematode morphology, taxonomy and phylogeny at Ghent University: a diversity of integrated toolkits.

Historically, the strength of nematology research at Ghent University lies within the field of morphology, taxonomy and phylogeny. From the backbone of nematode phylogeny to species delineation, we integrate today a tradition of light-microscopy and ultrastructural morphology with molecular approaches. We will provide an impression of our taxonomical work on free-living, plant-parasitic, virus-vector, facultatively parasitic, and entomopathogenic nematodes from natural and agricultural ecosystems of several continents. Our priority of coping with the enormous nematode diversity lies in integrative work in connection with other relevant research. In this presentation, we will focus on the following elements of our approach: ways to combine DNA sequencing data with morphology, the possibilities of electron microscopy, the assessment of intraspecific vs interspecific variation, applications of species delineation that are both theoretically sound and practically feasible, the integration of ecological and biological data, and the relevance to crop protection and biomonitoring. Finally we will discuss possibilities to integrate our data by the use of databases and phylogenetic frameworks. - 1Nematology Unit, Department of Biology, Ghent University, K.L. Ledeganckstraat 35, 9000 Ghent, Belgium. E-mail: [email protected]; 2Plant Protection Service, Nematology Section, Geertjesweg 15, 6706 EA Wageningen, The Netherlands; 3A.V. Zhirmunsky Institute of Marine Biology, FEB RAS, Vladivostok, Russia; 4Royal Belgian Institute of Natural Sciences, Department of Invertebrates, Vautierstraat 29, B-1000 Brussels, Belgium.

BLYUMMER A.G. & RIVKUS J.Z. Nematode infestation as an influential factor on the populations of Coptopsylla lamellifer rostrata Ioff et Tifl., 1934 (Syphonaptera, Coptopsyllidae) parasitising gerbils in the central and southern Kyzyl-Kum.

On the basis of material collected in central and southern latitudinal climatic subzones of the Kyzyl-Kum Desert in 1983-1997, it was shown that a large portion of female specimens of a flea, Coptopsylla lamellifer rostrata, parasitising the autumn-winter gerbil, was infected by nematodes. Maximum infestation of 32.2% was observed in the Central Kyzyl Kum during the 1st week of November. In the Southern part infestation was less extensive (20%), and occurred later on the same month. Distribution of Tylenchidae in the Kyzyl Kum appeared to have a focal nature. Within the boundaries of the area, blank (flea-free) zones of 1-6 km2 were detected. It was suggested that degradation of flea mezopopulations and C. l .rostrata elimination from the parasitocenosis of the great gerbil, occurred as a result of extensive nematode development and reaching the upper threshold infestation by nematodes in flea female specimens, respectively. - All-Russian Plant Quarantine Centre, Pogranichnaya, 32, Bykovo 140150, Moscow region, Russia. E-mail: [email protected].

BUTOVA K.B.1, 2 & PRIDANNIKOV M.V. 2 3 Plant-parasitic root-knot nematode Meloidogyne hapla.

Currently in the territory of Russia the in vivo data recorded only one species of root-knot nematodes that is relevant to agriculture: the northern root-knot nematode Meloidogyne hapla. The northern root-knot nematode has a wide host range, which in their natural habitat are the nature reservoir for the nematodes. Like other types of root-knot nematodes, M. hapla is an endoparasite of the root system. The cycle of the development proceeds in root tissues, and on its surface eggs sacs are formed. In the Central and North-Western areas one-two generations develop. In the place where females are feeding outgrowth of tissue (called galls) are formed, and sometimes several females form large (up to 1 cm in diameter) galls. The very typical reaction of the plants to infection by the northern root-knot nematode is the formation of numerous small lateral roots. The growth and development of the infested plants is delayed, and there is a significant deterioration in their commodity quality. In recent years major cases of infestation by this nematode in the Moscow and Pskov areas were noted. Meloidogyne hapla is an important subject for study from the point of view of plant protection, as there are no registered nematicides for open ground usage and there are no registered varieties of vegetable crops resistant to this root-knot nematode. - 1All-Russian Plant Quarantine Centre, Pogranichnaya str., 33, Bykovo, Moscow region, 140150, Russia. E-mail: [email protected]; 2All-Russian Research Institute of Phytopathology, Institute Str., Bolshie Vyazemy, Moscow region, 143050, Russia; 3Centre of Parasitology, A.N. Severtsov Institute of Ecology and Evolution, RAS, Leninskii pr. 33, Moscow, 119071, Russia.

CERMAK V.1, CUDEJKOVA M.2, MIKUSKOVA K.1, TOMANKOVA K.1 & GAAR V.1 Description of a Bursaphelenchus sp. (Nematoda: Parasitaphelenchidae) from peat substrate of hop seedlings from the Czech Republic.

In a phytosanitary survey conducted on hop seedlings produced in hop nurseries in the Czech Republic, a new species of Bursaphelenchus Fuchs, 1937 was extracted from peat substrate. This species is characterised by relatively short body lengths (female 395-481^m; male 417-510 ^m), short stylets (female 11.2-12.0 ^m; male 10.8-11.7 ^m) with weak basal thickening, small spicules (11.5-13.7 ^m) with minute cucullus, distinguishingly bent female tails with finely rounded terminus. The females have a small vulval flap. The species differs from the closest related species by morphometrics and morphological characters and also by species-specific ITS-RFLP profile. The sequence of the ITS region revealed, that this species is very closely related to Bursaphelenchus hofmanni Braasch, 1988. The new species has been successfully maintained in fungal cultures of non-sporulating forms of Botrytis cinerea for further studies. -1State Phytosanitary Administration, Division of Diagnostics, Slechtitelu 773/23, 779 00 Olomouc, Czech Republic. E-mail: [email protected]; 2C.R. Hana, Department of Molecular Biology, Slechtitelu 586/11, 779 00 Olomouc, Czech Republic.

CHIZHOV V.N., BUTORINA N.N. & PEREVERTIN K.A. Influence of a root parasitic nematode complex on productivity of fescue (Festuca pratensis Huds.) and timothy grass (Phleum pratense L.) in Central region of Russia.

Three-year studies of the numbers of root parasitic nematode species complex were conducted on crops of timothy grass (Phleum pratense L.) and fescue (Festuca pratensis Huds.), growing on medium loamy soils in the Central region of Russia. Dependence of grass productivity on free-living parasitic nematode population density, in general, without genus differentiation was observed. The relationship between estimated nematode numbers at the end of the second vegetation season and grass productivity on the third year of vegetation was the strongest. It was described by a classic exponential Seinhorst model. Parameters for two grass types and two yield forms, hay and seeds, were identified. The fact that productivity decrease of timothy grass was observed at lower parasitic nematode densities, compared with fescue, could be explained by different resistance of host plants, and differences in the qualitative composition of the parasite complex. Nematodes of the species Helicotylenchus digonicus predominated in the rhizosphere of both plant hosts. In the rhizosphere of timothy grass, Paratylenchus microdirus, Macropostonia sp. and P. pratensis were subdominant, and P. penetrans was noted. Paratylenchus microdirus and Macropostonia sp. were subdominant in the fescue rhizosphere. When the harmful effects of nematodes were assessed on the basis of seed mass, the maximum decrease in productivity of the studied grasses occurred at much lower nematode numbers, compared to assessments carried out on the basis of hay mass. - Center of Parasitology, IPEE RAS, Leninskii pr., 33, Russia, 119071, Moscow. E-mail: [email protected].

CLAEYS M.1, YUSHIN V.V.2, 3 & BERT W.1 High pressure freezing (HPF) and automatic freeze substitution (AFS): innovative techniques for transmission electron microscopy (TEM) in nematology.

The goal of morphologists, already at the beginning of electron microscopy, is to preserve the ultrastructure as close to the native state as possible. For the ultrastructural study of nematodes chemical fixation has several limitations.

Nematodes react quickly and this prevents penetration of the fixative resulting in ultrastructural artifacts. Hence, the worms have to be cut into pieces to facilitate penetration of chemical reagents. Here we present high-pressure freezing (HPF) in combination with automatic freeze substitution (AFS) as an alternative for classical chemical fixation. HPF has two main advantages compared to chemical fixation: a much faster rate of fixation and a simultaneous stabilisation of all cellular components. The technique is based on the suppression of ice crystal formation because of ultrarapid freezing (10000°C s-1) at a high pressure of around 2000 bar in liquid nitrogen. The nematode cuticle or even the eggshell is not a barrier for this fixation. The low-temperature substitution of dehydrating agents and fixatives into rapidly-frozen samples allows the crosslinking of cellular components and the removal of water at temperatures low enough to avoid the damaging effects of ambient-temperature dehydration. Taken all together, HPF followed by AFS shows superior morphological preservation for ultrastructural observations with good preservation of antigenicity for immunocytochemistry. Successful use of the technique is demonstrated, given our promising results on nematode gonads, gametes, spermatogenesis and localisation of major sperm protein. (Support: RFBR 11-04-00368; FEB RAS 12-III-A-06-098; RF 2010-220-01-180). - hematology Unit, Department of Biology, Ghent University, Belgium. E-mail: [email protected]; 2A.V. Zhirmunsky Institute of Marine Biology, FEB RAS, Vladivostok, Russia; 3Far East Federal University, Vladivostok, 690600, Russia.

COOSEMANS J. Saprophytic nematodes as bio-indivators insoil analyses.

Soil analyses to find out the causes for poor plant development are mostly based on chemical or biological data. Among biological data plant-parasitic nematodes receive the most attention. However, saprophytic nematodes contain important information as bio-indicators of the ecosystem. Based on numerous soil analyses, this study illustrates the benefits of including the total number of saprophytic nematodes and, even better, additionally in the analytical procedure, splitting up this group into colonisers and/or persisters or based on their trophic diversity.

Nematodes have many characteristics that make them suitable as bio-indicators: the high diversity of free-living terrestrial species, their abundance in the soil and the relative easy sampling methods. They are in close contact with the soil water solvents by their permeable cuticles and comprise heterogeneous trophic groups.

In general, the number of saprophytic nematodes fluctuates between equilibrium of 3000 to 5000 per 100 ml of soil. Numbers above and below this range provide strong indications that external elements have disturbed the ecosystem. Examples of soil analyses to find out the cause for poor plant development illustrate how the absence of saprophytic nematodes were the first indication of, for example, abundant use of fertilisers, pesticides or the use of immature compost or green manure. They are useful indicators for the efficiency of soil treatments against plant-parasitic nematodes. Related to the trophic groups, fungal and bacterial feeders, as opposed to the number of tylenchidae, are indicators of high biological soil activity. - Laboratory of Phytopathology, Dept. Biosystems KU Leuven, de Croylaan 42, B3001 Leuven, Belgium. E-mail: [email protected].

DABABAT A.A.1, ERGINBAS-ORAKCI G.1, BRAUN H.J.2, COMEZ H.2, DREISIGACKER S.2, MANNES Y.2, SINGH R.2, TOKTAY H.3, iMREN M.4, ELEKCIOGLU H.5 & MORGOUNOV A.1 Novel sources of resistance to cereal nematodes Heterodera spp. and Pratylenchus spp. in spring and winter wheat varieties.

Soil borne pathogens (SBPs), including the cereal cyst nematode (CCN), Heterodera species, and the root lesion nematodes, Pratylenchus species, attack roots of cereal crops and result in a high yield loss and reduced grain quality. The damage caused by these nematodes is accelerated in areas where drought exists. A few control options are being used to reduce CCN damage through keeping the population level below damage threshold, such as chemical, biological, cultural and genetic (resistance/tolerance) practices. However, up to now, resistance has only been identified against one of the CCN species, Heterodera filipjevi, in Turkey and foreign wheat germplasm though this resistance is not yet present in high yielding cultivars. Therefore, alternative approaches limiting the damage caused by CCN to wheat are needed. As a result of screening wheat germplasm against the CCN, hundreds of moderately resistant germplasm to H. filipjevi in winter wheat and to both Pratylenchus species and H. avenae in spring wheat germplasm are available. In 2012, germplasm with multidisease resistance including resistance to H. avenae, Pratylenchus thornei, P. neglectus and H. filipjevi were distributed to international collaborators. Two sets of spring and winter wheats are being genotyped to understand the genetic background of their resistance sources. - 1CIMMYT, P.K. 39 Emek 06511 Ankara, Turkey, 2CIMMYT Apdo. Postal 6-641, C.P. 06600, D.F. Mexico; 3Nigde University, Turkey; 4Biological Control Research Institute, Adana, Turkey; 5Cukurova University, Adana, Turkey.

DIYEVA D.S., MATVEEVA E.M. & SUSHCHUK A.A. Soil nematode communities of potato crop under heavy metal pollution.

Soil nematode fauna and community structure of functioning and former potato fields were studied. Soil samples were taken in the plots that are supposed to be polluted by heavy metals (Pb, Cu, Zn and Ni) with the wide range of concentrations (Petrozavodsk, Republic of Karelia). Functioning potato fields were characterised by the high quantity of nematodes (2500-3500 ind. 100 g-1 soil), a high proportion of plant parasites in the community structure (they were subdominant in the fauna) and the prevalence of nematodes of the genus Rhabditis (23-25%) with c-p-1 value (Bongers, 1990). The soil was infested by the specialised parasite of potato crops - Globodera rostochiensis Woll. Ecological indices (maturity index MI, channel index CI, structure index SI and enrichment index EI of soil food web) indicated the stable and developing soil food web with the inputting of large amounts of organic matter, which are decomposed by bacteria. Cessation of potato growing led to a restructuring of nematode communities (omnivorous nematodes became the subdominant trophic group; population density and index EI were reduced). Plots contaminated with heavy metal (Pb - 53 mg kg-1 soil, Cu - 87 mg kg-1) had plant parasites as the dominant eco-trophic group (40.9%) and a high percentage of Paratylenchus nanus in the fauna (21.1%). Productivity of potato crop was sharply reduced and the field was taken out of agricultural usage. Thus, the obtained data confirm the pattern, previously established by Suchchuk & Gruzdeva (2012) that soil ecosystem transformation connected with environment pollution led to increase in the number of plant parasites in the nematode fauna. Research was partially supported by Programme of Fundamental Research of Biology Department, RAS. - Institute of Biology of Karelian Research Centre of RAS, Pushkinkaya St., 11, Petrozavodsk, 185910, Russia. E-mail: [email protected].

EFEYKIN B.D.1, MIKHAILOV K.V.2, ALESHIN V.V.2, SPIRIDONOV S.E.1 & PANCHIN YU.V.2, 3 Phylogenetic relationships of nematodes and nematomorphs as inferred from sequence analysis of ribosomal proteins (RPL-RPS).

Nematomorphs or horse-hair worms are representatives of class Nematomorpha. All nematomorphs parasitise at the larval stage in the body cavity of arthropods. Adult worms escape from the host into water, where they copulate, lay eggs and give birth to tiny juveniles with a prominent proboscis. This latter penetrate the body of intermediate or definitive arthropod host. Phylogenetic relationships of Nematomorpha are still obscure: some authors consider these together with kinorhynchs and priapulids as members of the phylum Cephalorhyncha (Malakhov, 1980), whilst other consider nematomorphs as close relatives of nematodes (Schmidt-Rhaesa, 1996). Total DNA was extracted with the phenol-chloroform method from single specimens of Gordionus alpestris, collected in the stream in Western Caucasus. DNA was examined in Illumina Hi-seq 2000. Adapters and reads with low coverage were removed from the obtained reads. Contigs were assembled with Velvet. Two separate sequencing procedures provided an estimate of Gordionus alpestris genome as approx. 270 Mb. At least twentyfold coverage of the genome was obtained with N50 = 4050, which was sufficient for the RPL-RCF proteins. All the predicted RPL-RCF proteins were revealed in the obtained contigs. Before phylogenetic analysis of each separate protein, the ambiguous sites were removed from the alignment. Edited alignments were concatenated with ScaFoS 1.24. Maximum likelihood trees were constructed in RaxML 7.2.6. Conducted phylogenetic analysis supported the close relationships of Nematoda and Nematomorpha, with Tardigrada as the next closest relative. - 1Center of Parasitology, A.N. Severtsov Institute of Ecology and Evolution, RAS, Moscow, E-mail: [email protected]; 2M.V. Lomonosov Moscow State University, Belozersky Institute of Physico-Chemical Biology, Moscow, Russia; 3A.A. Kharkevich Institute for Information Transmission Problems, Russian Academy of Sciences, Moscow, Russia.

ESHOVA KH. & SAIDOVA SH.O. Influence of root-knot nematode Meloidogyne arenaria on the structure of tomato root tissue.

The present study is devoted to description of the peanut gall nematode Meloidogyne arenaria impact on the structure of tomato roots. It was established that correlative relation of physiological processes and cell and tissue differentiation in plant organs get disrupted. This results in formation of pathological morphologic structures, called galls. Giant multinuclear cells, cenocites and hydrocites are formed. - National University of Uzbekistan, 100099, A. Buriev Street, Tashkent, Uzbekistan. E-mail: [email protected].

FADEEVA N. & MORDUKHOVICH V. Metazoan meiofauna from north-western slope of the Sea of Japan with emphasis on free-living nematode assemblages.

Biota of the majority of the Sea of Japan slope areas is insufficiently explored; peculiarities of quantitative distribution of some of its groups and their geographical coverage are still not fully established. The present study was conducted to investigate the community structure and distributional patterns of meiofauna of the north-western slope of

the Sea of Japan. Sediment samples were collected during the Russian-German expedition SoJaBio (Sea of Japan Biodiversity Studies) on board of R.V. Akademik Lavrentyev in August 2010. Samples were taken from the depth ranging from 515 to 3367 m. In total, 22 quantitative samples from four sampling locations were analysed. Nematodes and harpacticoid copepods were the most abundant taxa. Meiofauna density values ranged from 45 ind. 10 cm-2 (water depth 3367 m) to 1620 ind. 10 cm-2 (water depth 515 m). In total, 125 free-living nematode species from 35 families were found in all samples. Number of species per sample varied from 14 to 22. Composition and structure of the nematode assemblages clearly changed with depths. Representatives of families Xyalidae (about 30%), Chromadoridae (about 20%) and Trefusiidae (about 20%) were predominant at the deepest locations. Density and composition of nematode assemblages at the genus level were comparable with those revealed at similar depths in waters around the world. Species diversity generally decreased with depth. - Far Eastern Federal University, Oktyabrskaya St., 27, Vladivostok, 690600, Russia. E-mail: [email protected].

GRUCMANOVA S. & HOLUSA J. Nematodes of double-spined spruce bark beetle Ips duplicatus (Sahlberg, 1836) from Central Europe.

This is one of the first studies on nematodes associated with Ips duplicatus in central Europe. Nematodes of this important pest species of bark beetle were studied on three localities. Infestatition by phoretic nematodes varied from 18 to 65%. Phoretic nematodes were found under elytra, on wings or between body segments, identified as Micoletzkya butschlii (Fuchs, 1915). Infestation by endoparasitic nematodes in the haemocoel of bark beetles varied from 3 to 30%. In haemocoel members of Contortylenchus sp. and Parasitylenchus sp. were found. In the intestine, juveniles of Parasitorhabditis obtusa (Fuchs, 1915) were found and its infestation rate was relatively low, from 0 to 16%. This species is the most abundant species found in Ips duplicatus galleries together with M. butschlii. And finally Cryptaphelenchus sp., Parasitaphelenchus sp. and some tylenchid juveniles were found in the gallery content too. -Faculty of forestry and wood sciences, Czech University of Life Sciences Prague, Kamycka 1176, 165 21 Praha 6 - Suchdol, Czech Republic. E-mail: [email protected].

GUZEEVA E.A.1 & YUSHIN V.V.2, 3 Structure and development of spermatozoa in the thelastomatid nematodes (Rhabditida: Oxyuridomorpha: Thelastomatidae).

The structure and development of spermatozoa in the thelastomatid nematodes, Hammerschmidtiella cristata and Leidynema portentosae, parasitising in the hindgut of the Madagascar hissing cockroaches, Gromphadorhina portentosa, were studied using transmission electron microscopy. The male reproductive system of thelastomatids is monorchic with reflexed testis. Spermatogonia are densely packed polygonal cells with rounded nucleus and distinct nucleolus; centrioles of 9 singlets are located near each spermatogonium nucleus; the cytoplasm contains mitochondria and ribosomes. Spermatocytes occupy almost two-thirds of testis length. The spermatocyte nucleus enlarges and chromatin is dispersed on inner surface of the nuclear envelope; electron-dense mitochondria surround the nucleus; the cytoplasm contains Golgi bodies and lipid droplets. Late spermatocytes bear numerous filopodia. Spermiogenesis starts with condensation of nuclear chromatin and fusion of mitochondria into dense cluster. The nucleus has no nuclear envelope. It is located posteriad to this cluster and becomes elongated. Immature spermatozoa (3*6-7 ^m) are elongated with widened anterior part and narrowed tail. The anterior part contains the cluster of mitochondria, a large electron-light body and numerous transparent vesicles of uniform size. Elongated and tapering nucleus protrudes into the tail-like posterior of the cell. All these peculiar characters of sperm morphology and development demonstrate the unique pattern of the thelastomatid male gametes within the order Rhabditida. (Support: RFBR 12-04-90847-mol_rf_nr; RFBR 11-04-00368-a; FEB RAS 12-III-A-06-098; RF 2010-220-01-180). - 1Centre of Parasitology, A.N. Severtsov Institute Ecology and Evolution RAS, Moscow, 119071, Russia. E-mail: [email protected]. 2A.V. Zhirmunsky Institute of Marine Biology FEB RAS, Vladivostok, 690059, Russia; 3Far Eastern Federal University, Vladivostok, 690600, Russia.

GUZEEVA E.A., KOSEVICH I.A. & SPIRIDONOV S.E. Molecular-taxonomic study of two pinworm species of the genus Pseudonymus (Oxyuridomorpha: Thelastomatoidea: Pseudonymidae) from water beetles (Coleoptera: Hydrophilidae) of the Astrakhan State Reserve.

Parasitism of thelastomatid nematodes in the hindgut of hydrophilid beetles was reported from different regions of the world. Currently, thelastomatids of water beetles constitute the family Pseudonymidae. Taxonomic relationships of separate genera and species within this family are still not completely resolved, and additional characters for species identification are urgently needed. Two species of the genus Pseudonymus Diesing, 1857 were collected from the hindgut of a hydrophilid beetle (Hydrophilus piceus L.) sampled in the River Volga delta. Two species of thelastomatids were recovered, identified according to morphological features as Pseudonymus spirotheca (Gyory,

1856) and P. islamabadi (Basir, 1941). Pronounced sexual dimorphism prevents definite identification of con-specific males and females, and previously such judgments were based solely on comparative occurrence of the specimens of both sexes in dissected hosts. Sequences of the D2-D3 segment of the LSU rDNA for two morphological types of males and females were obtained and compared (alignment length 636-637 bp; nucleotide difference between two species 44 bp). Complete identity of LSU sequences confirmed that females with only one widened annulus and 70*40 ^m eggshells and males with narrowed tail with short spine appendage and 25 ^m long spicule belong to P. spirotheca, whereas females with strongly swollen first, fifth and seventh cuticular annuli of the anterior end and 85*45 ^m eggshells and males with rounded posterior end and spicule length of 27 ^m belong to P. islamabadi. Morphological features of two Pseudonymus species from lower Volga are illustrated with SEM. - Centre of Parasitology, A.N. Severtsov Institute Ecology and Evolution RAS, Moscow, 119071, Russia. E-mail: [email protected].

HANDOO Z.A., MOWERY J.D. & CHITWOOD D.J. The history of the USDA NEmatode collection and its DAtabase: valuable resources for taxonomic research and identification.

The USDA Nematode Collection continues to serve as one of the largest and most comprehensive nematode repositories in the world. Nematology research began in the United States in the late 1800s, and for many years the collections of the USDA nematologists remained as individual personal collections, and unfortunately over those years many valuable specimens deteriorated or were misplaced and collection records lost. The process of salvaging the surviving specimens of previous nematologists remains an ongoing effort. In 1960, with type specimens of 18 species, A. Morgan Golden officially established the USDA Nematode Collection at Beltsville, Maryland, creating an organised, unified repository of important specimens. The oldest slide in the Collection is of Mononchus longicaudatus, from Australia, that was prepared by Nathan A. Cobb in 1890. Today, the USDA Nematode Collection is comprised of several constituent divisions, which collectively consist of over 45,000 slides and vials. Essential data is recorded for each species and all depositions are entered into a database in which over 39,000 entries are searchable and available to the public at http://nt.ars-grin.gov/nematodes/search.cfm. These resources serve as a major asset for taxonomic research and are used for a wide variety of scientific and regulatory purposes, including the resolution of billion-dollar issues involving agricultural trade, the continued protection of agriculture against economically dangerous invasive species and the advancement of nematode taxonomy and scientific research, including nematode identifications. Scientists around the world have regularly incorporated specimens from the Collection into their research and are encouraged to deposit material into the Collection. - USDA ARS Nematology Laboratory, 10300 Baltimore Ave., BARC-West, Bldg 10A, Beltsville, MD, 20705, USA. E-mail: [email protected].

IESHKO E.P., MATVEEVA E.M. & SYSOEVA M.I. Genotype-related variation in potato susceptibility to

Globodera rostochiensis.

Susceptibility of four potato genotypes derived from the wild Solanum commersonii Dun. was studied. Genotypes differed in freezing tolerance to the strictly specialised potato cyst nematode (PCN) Globodera rostochiensis (20 cysts plant-1). The highest susceptibility to the infestation was observed in the wild species and the highly freezing-tolerant (FT) genotype. A much lower nematode infestation rate was found in the moderately freezing-tolerant genotype. The highest resistance to PCN was observed in plants of the freezing-sensitive (FS) genotypes and the cultivated species S. tuberosum (cv. Nevsky). To check whether there had been cross-adaptation caused by exposure of the plants to low temperatures prior to the infestation; all genotypes were cold-pretreated. Resistance to the nematode infestation was found to be enhanced, as seen in the lower number of cysts, and the profoundness of the effect was inversely proportional to the plant response to low temperature (FT and FS genotypes). The reduction in the nematode infection in the wild species and highly FT genotype after exposure to low temperatures was negligible. Although there was a trend for reduction (by 12-28%), the differences in the number of cysts were statistically insignificant. The moderately FT genotype displayed a statistically significant reduction in the rate of the nematode abundance after cold treatment (the nematode reproduction coefficient к was 1.4 vs. 4.9 in the control). After being cold-treated all FS potato genotypes and the cultured species demonstrated higher resistance to the nematode infestation (к = 1.2-2.2 vs 2.6-4.0 in the control). The differences between treated and control plants were statistically significant (Р < 0.05). - Institute of Biology, Karelian Research Centre, Russian Academy of Sciences, 11 Pushkinskaya St., 185910 Petrozavodsk, Russia. E-mail: [email protected].

JAMALI S.1 & SHOKOOHI E.2 Nematode antagonistic activity of medicinal plants aganist Meloidogyne

javanica.

Among different phytoparasitic nematode management strategies, chemical control has proved generally effective but the majority of these chemicals are expensive and often hazardous. Biological control seems to be an alternative

management practice for plant-parasitic nematodes. Plant extracts possess nematicidal properties and their potential use in nematode control programmes has been proved effective. Nematicidal properties of leaf extract of medicinal plants including Datura fastuosa, Albizia julibrissin, Acacia leucophloea, Amaranthus caudatus, has been carried out to evaluate their potential toxicity against root-knot nematode, Meloidogyne javanica, on tomato in laboratory and greenhouse conditions. Experiments were performed with leaf extract using randomised complete block design. The dead nematodes were observed under stereoscopic binocular microscope after 24 to 48 and 72 h and percentage mortality was calculated. Nematodes were considered dead if they did not move when probed with a fine needle. The results revealed the toxicity increased with an increase in concentration as well as the exposure time. The results also indicated that most of the tested treatments obviously reduced root galls and increase tomato plant growth characters significantly and egg masses on root system, as well as the numbers of juveniles in the soil. Overall, Datura fastuosa leaf extracts gave the maximum nematode mortality (78%) followed by Albizia julibrissin (65%), Acacia leucophloea (42%) and Amaranthus caudatus (38%). Use of leaf extracts showed a maximum increase in growth of tomato plants, which was found to be associated with increasing concentration of extracts and subsequent decrease in nematode population. - 1Plant protection Department, College of Agricultural science, University of Guilan, Rasht, Iran. Email: [email protected]; 2 Plant Protection Department, College of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran.

JANSSEN T.1, KARSSEN G.2, COUVREUR M.1 & BERT W.1 Phylogeny, taxonomy and evolution of reproduction within the nematode genus Pratylenchus: a multi-gene approach combined with karyotyping.

Root-lesion nematodes of the genus Pratylenchus (Filipjev, 1936) are an important pest in economic crops (Sasser & Freckman 1987). They have a worldwide distribution (except for polar regions) and can parasitise a very wide range of plants. At the moment the genus includes about 70 described species, but identification of the different species based on morphology remains problematic. Within the genus reproduction strategies vary from amphimixis through meiotic to mitotic parthenogenesis. Also karyotypes are highly variable and polyploidy is certainly present and probably associated with asexual reproduction (Roman & Triantaphyllou, 1969). The current study is based on as many species as possible; sampling will be done at type locations if possible, and all populations are studied from multiple and complementary perspectives. Morphology is documented using light and electron microscopy and mode of reproduction and karyotypes are elucidated by in vitro staining of reproductive systems. This information will be analysed in a multigene comparative evolutionary perspective to obtain a better insight into the evolution of reproductive strategies, genome evolution and phylogenetic relationships. The obtained data and outcomes will be highly valuable towards a complete integrative taxonomy of the genus Pratylenchus; initial results will be presented here. - hematology Unit, Ghent University, Department of Biology, Ledeganckstraat 35, 9000 Ghent, Belgium. E-mail: [email protected]; 2Plant Protection Service, Nematology section, P.O Box 9102, 6700 HC Wageningen, The Netherlands.

KHUDYAKOVA E.A. & SUDARIKOVA S.V. Validation of the FLASH-PCR method for identification

Globodera rostochiensis (Woll.) Behrens.

A research on recognition of reliability (validation) of cyst nematode diagnostics employing a commercial diagnostic set produced by the company Agrodiagnostika (Moscow) was conducted. The following indicators of validation were defined according to the EPPO PM Standards 7/84, RM 7/98: 1. Analytical sensitivity. In a series of experiments it was shown that the detection limit of viable cysts was at least five juveniles or eggs per sample, and not less than 50 juveniles or eggs per sample for non-viable cysts. 2. Analytical specificity. As a result of the experiment, a non-specific reaction of the employed primers with Globodera pallida and Heterodera sp. was observed. Together with this, a specific reaction with Globodera rostochiensis was observed. The experiment revealed high specificity of the employed set for FLASH-PCR to Globodera rostochiensis. 3. Repeatability. The study was conducted on one and the same day by one and the same operator employing one and the same equipment. The experiment showed a significant repeatability 100%. 4. Reproducibility. Results of this experiment also enabled Globodera rostochiensis to be identified in all samples, which indicate high reproducibility of the method. As a result of the study, we conclude that the molecular FLASH-PCR method is suitable for identification of a cyst nematode, Globodera rostochiensis (Woll.) Behrens, in diagnostic quarantine laboratories. - All-Russian Plant Quarantine Centre, 140150, Bykovo, Moscow region, Russia. E-mail: [email protected].

KHUSAINOV R.V. Fauna and ecology of wood-inhabiting nematodes from brittle willow (Salix fragilis) in the Central European Part of Russia.

Fauna and ecology of wood-habiting nematodes dwelling in brittle willow (Salix fragilis) and their relation to various wood life cycles were studied in 2010-2012. Samples were collected in six regions of the Central European Part of Russia. They were divided into six types based on three main parameters such as rind surface structure, sanitary state of wood and presence of simple or complicated epiphytes. Consequently representatives of 32 genera from 18 families belonging to 8 orders were detected. Phleobionts (algaephages and bacteriophages) were dominated in strong wood without any mechanical destruction (type A and type B). Dorylaimids and plectids were also present in wood that had been extensively covered by complicated epiphytes (moss and lichen) (type C). There were many various mycetophages from Aphelenchida and Tylenchida orders in glabrated wood (type D). Saprobionts and edaphobionts predominated in perished and decayed wood (types E and F). In general, entomopathogenic species were discovered only in both of the last types of wood, but their quantities differed markedly. - Center of Parasitology, A.N. Severtsov Institute of Ecology and Evolution, RAS, Leninsky prospect 33, Moscow, 119071, Russia.

KOLESOVA E.A. & SHESTEPEROV A.A. Development of a computer model "Forecast yield loss of potato crop depending on the intensity of Globodera infestation".

The mathematical models of potato yield loss forecast for two varieties of potato, Sineglazka and Nevsky, were worked out. The first models demonstrated the potato yield loss depending on the population density of golden potato nematode (the coefficient of determination R2 = 0.303, R2 = 0.326); the second models showed it depending on the intensity of Globodera infestation (the coefficient of determination R2 = 0.68, R2 = 0.76). The second models were more mathematically sound. Inverse proportion was observed between potato yield and intensity of Globodera infestation. The dependence of the potato yield on intensity of Globodera infestation development could be mathematically described by the formula: y = b - ax, where b - coefficient, which depends on the variety of features, x - intensity of Globodera infestation in percentage. Imitational experiments involving the interactive computer model revealed that the intensity of Globodera infestation development (from 10 to 90%) caused yield losses of Sineglazka potato variety (from 12.2 to 78.2%), and Nevsky variety (from 10.8 to 64.9 %). - Russian State Agrarian External University, Balashicha, 143900, Russia. E-mail: [email protected].

KOOHKAN M. & SHOKOOHI E. Preliminary study for mass production of Mylonchulus sp. in laboratory condition.

Predatory nematodes play a vital role in food web ecology. Mylonchulus is a widely distributed predatory soil nematode and is of interest in the study of soil ecology, yet very little information exists on its in vitro culturing. In this investigation, an artificial environment (SEA) was created to maintain Mylonchulus for mass productional studies. One hundred prey nematodes (i.e., Tylenchus, Aphelenchus, Helicotylenchus and Ditylenchus) were added as food sources. The Petri dishes were maintained at 25°C in the dark. Five adults were added to each Petri dish. The number of Mylonchulus was recorded 54, 60 and 65 days after nematode inoculation to the medium. The first juveniles were observed after 56 days; they increased to 10 after 60 days, and after 65 days their number increased to 15. The results revealed that juveniles of Mylonchulus could not survive without adults in cultured medium and, therefore, the adults may prepare the prey for juveniles. Furthermore, a thin layer of water is needed for activation and mass production of Mylonchulus in the medium. - Plant Protection Department, College of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran. E-mail: [email protected].

KORMA A.M. Biological peculiarities of Bursaphelenchus nematodes in East Polesia, Ukraine.

Research of wood nematode fauna enabled the composition of Bursaphelenchus nematodes species in Eastern Polesia, Ukraine to be determined. The nematodes are represented by three species: Bursaphelenchus mucronatus Mamiya & Enda, Bursaphelenchus sexdentati Ruhm, and Bursaphelenchus eggersi Ruhm. Bursaphelenchus mucronatus species predominate among xylobiont nematodes and have detection frequency of 54.1%. Some ecological peculiarities have been established of the area of distribution of B. mucronatus being enlarged due to insect-carriers not described previously in the literature. The author discusses the ascertained ecological peculiarities and domination of B. mucronatus species. The research demonstrated the transfer of Buprestis mucronatus nematodes by coniferous Buprestis octogutta L. (Ancylocheira octoguttata L.). - Chernigov State Institute for Economics and Management, 1 Streletskaya St., Chernigov, 14033, Ukraine. E-mail: [email protected].

KOROPETS S.I. Nematodes of Scotch pine seedlings: diagnostics, peculiarities of development and the forecast.

The complex of plant nematodes associated with Scotch pine (Pinus sylvestris L.) in Ukrainian forestry nurseries was investigated. We detected 67 species of plant nematodes. According to the ecotrophic characteristic, all species belong to four groups: phytohelminths (17 species), mycohelminths (9 species), saprobic (36 species) and predatory (5 species). Phytohelminths Coslenchus costatus, Aglenchus agricola, Pratylenchus vulnus, Ditylenchus dipsaci; mycohelminths Aphelenchoides asterocaudatus, Aph. minimus and saprophages Acrobeloides buetschlii, Cervidellus insubricus were common to the rhizosphere of healthy seedlings. Peculiarities of nematodoses development and problems of their monitoring are discussed. - National University of Life and Environmental Sciences, Heroyiv Oborony St., 15, Kyiv-03041, Ukraine. E-mail: [email protected].

KOSAKA H.1, SAYAMA K.2, KANZAKI N.3 & MAKINO S.4 Host range and geographical distribution of Sphaerularia vespae, a parasite of hornets.

Sphaerularia vespae has been found in a common Japanese hornet, Vespa simillima, and described as the second species of the genus Sphaerularia. Another species of this genus is S. bombi, a well-known parasite of bumblebees. As is the case with S. bombi, potential queens of V. simillima parasitised by S. vespae are sterilised. To understand the basic ecology of S. vespae, its host range and geographical distribution were investigated. Sphaerularia vespae was found in four of the seven species of hornets (Vespa spp.) native to Japan: V. simillima, V. mandarinia, V. dybowskii and V. ducalis. In V. analis, only immature S. vespae were found but no mature nematodes were detected. This suggests that V. analis is not a host of S. vespae and that S. vespae cannot complete its lifecycle in V. analis. Sphaerularia vespae was not found in V. crabro and V. affinis. Sphaerularia vespae was found in Hokkaido, Honshu and Kyushu. In Hokkaido the prevalence of S. vespae was highest in V. simillima, while in Kyushu it was highest in V. mandarinia; both are dominant hornet species in their respective regions. These results suggest that S. vespae is widely distributed across the main islands of Japan from Hokkaido to Kyushu. However, S. vespae has not been found in Iriomote Island, located almost at the southwest end of Japan, or in Korea. Further research is necessary to confirm host adaptation of S. vespae to V. crabro and V. affinis and its distribution in the southern islands of Japan and other countries. - 1Kyushu Research Center, Forestry and Forest Products Research Institute, Kurokami 4-11-16, Kumamoto 860-0862, Japan. Email: [email protected], 2Hokkaido Research Center, Forestry and Forest Products Research Institute, Hitsujigaoka 7, Sapporo, Hokkaido 062-8516, Japan; 3Department of Forest Microbiology, Forestry and Forest Products Research Institute, Matsunosato 1, Tsukuba, Ibaraki 305-8687, Japan; 4Forestry and Forest Products Research Institute, Matsunosato 1, Tsukuba, Ibaraki 305-8687, Japan.

KOSHELEVA T.N. Morphological anomalies of free-living marine nematodes of the Black Sea.

New data concerning morphological anomalies of nematodes found in the coastal zone (Sevastopol region) and the oxic/anoxic interface (NW Crimea and the Bosporus region) of the Black Sea have been discussed. The structure and localisation of amphid are species-specific features of free-living nematodes. Normally the amphid is a paired organ having certain localisation in the anterior part of body. Previously (Sergeeva, 1991, 2003) it was noted the following spectrum of amphid anomalies: 1) the number of amphids could be 1-5 instead of two in norm; 2) the form of amphids could be different; 3) one of two amphids in specimens could be underdeveloped; 4) one of two amphids could be divided into parts; 5) amphid location in relation to the anterior edge and to the axis symmetry of the body is inconsistent. Our new data show that the number of Black Sea nematodes manifesting such deviation, has increased from 13 to 33 species. Most often we found anomalous specimens in the following populations: Terschellingia longicaudata (0.6-100%), Axonolaimus setosus (2.4-100%), Parodontophora quadristicha (2.9-100%). The present study, as well as previous ones, showed that such spectrum of abnormality in the development of amphids of the Black Sea nematodes is not an accidental phenomenon. Obviously, anomalous features may be hereditary because they are found in the same species in females, males and young specimens.Accumulated data concerning the anomalous development of nematodes in various water areas of the Black Sea may indicate the reaction to some adverse environmental conditions and should be observed in the other waters as well. - Institute of Biology of the Southern Seas, NAS Ukraine, 2, Nakhimov Av., Sevastopol, 99011, Ukraine. E-mail: [email protected].

KUDRIN A. & LAPTEVA E. Natural flooding effects on soil nematodes communities.

Floodplains are unique ecosystems that consist of two subsystems, an aquatic and a terrestrial one. Inundation of a floodplain can be a stress to the both aboveground and belowground biota. Soil nematodes are protoaquatic organisms. They demonstrate a high resistance to environmental extremes through resistance adaptation and other types of survival strategies, and, probably, can be resistant to the spring/summer flood. However, no data on the response of soil

nematodes to natural flooding is available. Our objective was to examine the effects of flooding duration on nematode in the floodplain. The study was conducted on floodplain forests of the Pechora and Sisola River, in taiga zone of northwest Russia. At each floodplain three sites with various flooding time were evaluated: (1) brief flooding, 2-3 days; (2) moderate flooding, 2-3 weeks; (3) extended flooding, 8-12 weeks. Our results are strongly indicative of a negative impact of natural flooding on soil nematodes. They clearly correlate with the general trend, according to which abundances and biomass of soil invertebrates are reduced immediately by flooding. The effect becomes more pronounced with the increase in flooding duration, but even then, it is reversible and is usually compensated to a large extent during the next soil-dry period. One of the major reasons of negative effect is probably a decrease of oxygen concentration in the soil that has direct and indirect influence on nematodes. - Institute of Biology, Komi SC, RAS, Kommunisticheskaya St., 28, Syktyvkar, Russia. E-mail: [email protected].

KULINICH O.A.1, 2, ARBUZOVA E.N.2, KOZYREVA N.I.2, MAZURIN E.S.2, ROMASHOVA N.B.2, KOLYCHIKHINA M.S.2 & RYSS A.YU.3 Bursaphelenchus mucronatus nematodes and their associated bacteria as a possible cause of the pine forest death in Russia.

During the survey of conifer forests in Russia (2010-2012), twenty-four isolates of the wood-inhabiting nematode Bursaphelenchus mucronatus were extracted and propagated in vitro to determine the symbiotic bacteria associated with them. Sixteen species of bacteria belonging to nine families Enterobacteriacea, Xanthomonadaceae, Pseudomonadaceae, Burkholderiaceae, Rhizobiaceae, Nocardiaceae, Flavobacteriaceae, Bacillaceae and Paenibacillaceae were isolated from the nematodes and identified, i.e., Achromobacter, Bacillus, Burkholderia, Enterobacter, Flavobacterium, Klebsiella, Pseudomonas, Rahnella, Rhodococcus, Stenotrophomonas, Pantoea, Paenibacillus and Serratia. The bacterium Pseudomonas fluorescens was isolated from nine B. mucronatus isolates. According to the results of Chinese researchers, P. fluorescens is an essential species of the nematode-bacterial complex that induces Pine Wilt Disease in pine forests of southern China. Thus, we can make an assumption that in 2010; B. mucronatus nematodes and symbiotic bacteria of P. fluorescens caused the death of some pine forests in areas where the mean air temperature in European Russia was 26.4°С in June and 25.5°С in August. - 1Center of Parasitology, A.N.Severtsov Institute of Ecology and Evolution, RAS, Leninsky Prospekt 33, Moscow, 119071, Russia. E-mail: [email protected]; 2All-Russian Plant Quarantine Centre, 140150, Moscow region, village Bykovo; 3Zoological Institute RAS, St. Petersburg, 199034, Russia.

LAVROVA V.V., SYSOEVA M.I. & MATVEEVA E.M. Temperature priming is a base of enhanced plant resistance to phytonematode.

A potato cyst-forming nematode (PCN) Globodera rostochiensis Woll. is one of the harmful parasites of the potato crop. Plants can acquire enhanced resistance to pathogens after treatment with synthetic or natural compounds (for example, p-aminobutyric acid and chitosan). The induced resistance is associated with an enhanced capacity for the rapid and effective activation of local and systemic defence responses. This process is called 'priming'. Priming can be mediated by temperature, because the latter modulates plant immune responses. The aim of the study was to investigate potato plant responses to short-term low temperature impact before nematode infestation. Experiments were conducted in the growth chamber and the field conditions on seedlings and tuber respectively. Resistance formation to PCN, level of resistance gene expression, activity of photosynthetic apparatus and fatty acid content were analysed. Results showed that independently of treatment period (tubers or seedlings), a temperature drop decreases nematode infestation level and promotes plant fitness. DROP-treated potato contained more polyunsaturated fatty acids. These plants also expressed elevated level of nematode resistant gene (Hi). Thus, short-term temperature drop may lead to the induction of a primed state of potato plant that sensitises the tissue for improved elicitation of various defense responses during nematode invasion. Plant temperature priming may be a result of histone modifications and chromatin remodeling. Research was supported by Ministry of Education and Science of Russian Federation (project 8050). - Institute of Biology of Karelian Research Centre RAS, Pushkinskaya St., 11, Petrozavodsk, 185910, Russia. E-mail: [email protected].

MATVEEVA E.M., SUSHCHUK A.A. & DIYEVA D.S. Soil nematode communities under annual and perennial crops.

Permanent (common) and variable (specific) components of nematode fauna under annual (potato, cabbage, carrot and beet) and perennial (seeded meadow with single species Phleum pratense L.) were determined. Nematode fauna under alien species Heracleum sosnowskyi Manden was compared to nematode communities of different types of agrocenoses and plants introduced and naturalised in biocenoses. Taxonomic diversity (16 genera) and nematode quantity (250 ind. 100 g-1 soil) were low under annual crops, especially carrot and beet, but high under P. pratense and

H. sosnowskyi (27 genera, 1241-5125 ind. 100 g-1 soil. Common taxa for annual crops according to semi-taxonomic feeding classification belonged to Rhabditida r-and k-strategic bacterial feeders (7 genera), Adenophorea bacterial feeders (1), Tylenchida fungal feeders (3) and Dorylaimida omnivores (2). Under annual crops, maturity, channel and structure indices values were low and enrichment index was high. Therefore, regular use of agricultural techniques led to domination of bacterial feeders with c-p = 1 and simplification of community structure. Soil ecosystems were assessed as disturbed. In biocenoses with P. pratense and H. sosnowskyi MI and CI were higher and EI was lower. Introduction and subsequent establishment of alien species in plant community (at the edge of potato crop) did not lead to negative consequences for nematodes during 10 years. Nematode fauna stayed similar to that of potato crop in species composition but differed by occurrence of Adenophorea bacterial feeders uncommon for agrocenoses. Low SI (16.3) indicated degraded food web and stressed state of soil ecosystem. Research was partially supported by Programme of Fundamental Research of Biology Department RAS. - Institute of Biology, Karelian Research Centre, Russian Academy of Sciences, 11 Pushkinskaya St., 185910, Petrozavodsk, Russia. E-mail: [email protected].

MCGAWLEY E.C., OVERSTREET C. & PONTIF M.J. Variation in reproduction and pathogenicity of geographic isolates of Rotylenchulus reniformis on cotton in America.

The comparative reproduction and pathogenicity of isolates of Rotylenchulus reniformis from the American states of Alabama, Arkansas, Hawaii, Louisiana, Mississippi and Texas on cotton was evaluated in microplot trials. Prior to initiation of microplot trials, ten single egg mass (SEM) populations of each geographic isolate were derived from single egg masses. Reproduction of the SEM populations of each geographic isolate were evaluated in greenhouse studies with Stoneville LA887 cotton by assessing the numbers of vermiform stages in soil and eggs per gram of root tissue 60 days after inoculation. On the basis of these trials, each repeated once, one SEM population of each of the six isolates was selected for use in microplot trials. Averaged over the two trials, SEM population designations selected for use in microplot trials and their respective reproduction values (R, where R = Pf/Pi) and numbers of eggs g-1 root were: AL-8 (R = 14.9, eggs = 202); AR-3 (R = 30.4, eggs = 525); HI-9 (R = 20.2, eggs = 183); LA-3 (R = 18.2, eggs = 517.); MS-7 (R = 25.7, eggs = 602) and TX-10 (R = 42.8, eggs = 938). Data from full-season (147 days) microplot trials, averaged over 2 years, showed significant differences (Tukey's HSD test (P < 0.05%)) among isolates of reniform nematode in both reproduction and pathogenicity. Dry plant weight at harvest averaged 370.6 g for the non-inoculated control. All isolates except the one from HI produced root weights at harvest that were reduced significantly below that of the control. Harvest weights for plants inoculated with LA-3 and MS-7 were significantly lower than those from the other four geographic regions. - Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA. Email: [email protected].

MEHDIZADEH S.1 & SHOKOOHI E.2 Seasonal distribution of free-living and plant-parasitic nematodes in the rizosphere of apple in Kerman province of Iran.

The seasonal distribution of the free living (rhabditids) and plant-parasitic nematodes (tylenchids) were investigated over a 1-year period. Samples were collected at depths of 35-40 cm, on clay soil of the rhizospheres of apple trees in November, March, May and July of 2012. Nematodes numbers ranged from 2 to 63 individuals 10 cm-2 soil. Enzyme Commission number (EC) and pH features were determined. The pH ranged from 7.3 to 8.02 in different seasons. The total nematode density was low during November (autumn) (24 individuals 300 cm-2) and reached a maximum number of 223 individuals 300 cm-2 soil in March (winter). The results indicated that population dynamics of nematodes had a direct relationship with EC and pH. Therefore, if pH increases, density of plant-parasitic nematodes increases, while density of free-living nematodes decreases. On the other hand, the results of pH indicate that when pH is 8.2, the plant-parasitic nematodes have maximum density. In this condition, the genus Pratylenchus has maximum density (March, pH = 8.2). However, in Ph = 7.50 the genus Panagrolaimus reaches maximum density (44 individuals 300 cm-2 soil). Furthermore, distribution of the genera depends on the season. The results showed that the genus Acrobeles was found only in spring (May). However, the most frequent nematodes were Helicotylenchus, Pratylenchus, Aphelenchus, Panagrolaimus and Mesorhabditis, which were found in all seasons. - 1Young Researchers Society, Shahid Bahonar University of Kerman, Kerman, Iran; 2Department of Plant Protection, College of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran. E-mail: [email protected].

MIGUNOVA V.D. & LYCHAGINA S.V. Nematodes associated with vegetable cultures in greenhouses of Saratov region.

The analysis of rizosphere soil of three greenhouses in Saratov region for the presence of plant-parasitic and free-living nematodes was performed. The analysed territory total area was 3 ha. The total number of nematodes was 392 ±

48 specimens 100 ml-1 soil. Average number of Meloidogyne incognita juveniles was 120 100 ml-1 soil. Other genera were represented by Acrobeloides, Rhabditis, Acrobeles, Aphelenchoides, Dorylaimus and Plectus. A four-fold increase of the number of predacious nematode Butleriellus sp. and presence of dorylaimids led to the decrease of numbers of root-knot nematode juveniles in the nematode-complex (from 32 to 19%). This might be related to regulatory impact of predators under greenhouse conditions. Thus, the fauna of predacious nematodes of greenhouses is poor. Nematodes of the family Mononchidae were absent in greenhouse substrate. The dorylaimid nematodes were rare and few. The predacious nematodes of genus Butlerius (fam. Diplogasteridae) were frequently found in greenhouse substrate. This nematode genus may be considered a potential agent of biological control of plant-parasitic nematodes under greenhouse conditions. - All-Russian K.I. Skryabin Scientific Research Institute of Helminthology, Bolshaya Cheremushkinskaya 28, Moscow, 117218, Russia. E-mail: [email protected].

MIRONENKO N.V.1, ROGOZINA E.V.2, LIMANTSEVA L.A.1 & AFANASENKO O.S.1 Fitness of pathotype Ro1 Globodera rostochiensis on partially resistant potato hybrids.

An interaction between the golden nematode Globodera rostochiensis (Woll.) Behrens pathotype Ro1 and potato has been studied by using partially resistant potato hybrid clones, which are the offspring of wild species native to South America. A few cysts formed on the roots of partially resistant plants were used for re-infection of the same genotypes of potato interspecific hybrids. Reproductive ability of the nematode second generation after two inoculations of 12 hybrid clones was measured by the index of reproduction and the number of second-stage juveniles (J2) per cyst. On six potato hybrid clones the indices of reproduction in the second generation were evaluated as "0", so there was no propagation of G. rostochiensis. On the other six potato clones we observed the suppression of nematode propagation. Reproductive ability was lower than on the roots of cv. Nevsky: the index of reproduction estimated 0, 5-4 and 69 versus 10, 33 and the number of J2 per cyst estimated on average 34 vs 67 pcs. The proportion of nematode cysts having a small number of J2 (30 pcs) in a second generation progeny collected from hybrid clones was three times more than from cv. Nevsky. We found single large cysts (5% and 1%) with 80-160 J2 after a second inoculation on hybrids with S. doddsii and S. incamayoense in their pedigree. On cv. Nevsky, the number of large cysts was determined as 1722%. Our results show that the initial soil-borne population of G. rostochiensis is heterogeneous. Depending on the genotype of the host plant, selection among hatched J2 has occurred. - 1All Russian Institute for Plant Protection, Pushkin, shosse Podbelskogo, 3, St. Petersburg, 196608, Russia. E-mail: [email protected]; 2N.I. Vavilov Research Institute of Plant Industry (VIR), 42-44, B.Morskaya Street, 190000, St. Petersburg, Russia.

MLADENOV A., LAZAROVA S., ELSHISHKA M., MINCHEVA Y. & PENEVA V. Impact of ski runs on soil nematode assemblages from different habitats on North Pirin Mountain - preliminary results.

Soil nematode assemblages of different habitats (Mountain (Pinus mugo L.) and Macedonian Pine (Pinus peuce Griseb.) and spruce (Picea abies L.) forests, subalpine meadows and ski runs - bare and with grasses) are to be studied in the frame of a PhD project in order to assess changes caused by ski runs in various community parameters based on newly collected material (2009) and a previous study (2005). The results on structural and functional diversity of nematode assemblages of Macedonian Pine forest fragments (8 sites) and ski runs (4 bare ski runs and 5 runs with grasses) are presented and discussed. Several ecological parameters were analysed: relative abundance, diversity and maturity indices, trophic and life strategies functional group. Nematode assemblages of bare runs formed a separate group, characterised by a very low abundance and generic richness and composed of genera belonging mainly to c-p2 and c-p4 groups, diverse trophic structure (with a very low percentage of plant feeders); ski runs with grasses supported the most abundant and diverse nematode assemblages, dominated by bacterial feeders and omnivorous nematodes, again c-p2 and c-p4 prevailing; Macedonian Pine fragments had relatively balanced trophic structure of nematode communities, with plant feeders having a very small proportion and generic richness compared with grass communities; sites studied in 2009 had substantially lower number of genera and abundance. Two of bare ski runs sampled in 2005 were sampled again in 2009; however, currently they represent ski runs with grasses and thus the strongly degenerated soil nematode assemblages have partly recovered. - Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences, 2 Y. Gagarin Street, 1113 Sofia. E-mail: [email protected].

NADI M.1, SHOKOOHI E.1 & TROCCOLI A.2 Molecular analysis of Helicotylenchus cf. digonicus from south of Iran based on sequence of the 28S ribosomal DNA.

Helicotylenchus is a cosmopolitan genus including some important plant-parasitic nematodes causing damage in agricultural and horticultural crops. During a soil sampling of almond (Prunus) orchards in Baft region of Kerman province (South East of Iran), a species of Helicotylenchus was extracted from the rhizosphere of almonds and identified according to morphological and morphometric characters as H. digonicus. In addition, the D2D3 segment of

28S rDNA was amplified using specific primers and sequenced. The Nblast result showed that the studied population has 8 nucleotides differences with Italian population of H. digonicus (DQ328758; 99% identity). Phylogenetic analysis using Maximum Likelihood places this population close to an undescribed population of Helicotylenchus (HM014303) and H. digonicus. Genetic pairwise distances showed low variation (0.002) with a population of H. digonicus (HM014241) from South Africa. - 1Plant Protection Department, College of Agriculture, ShahidBahonar University of Kerman, Kerman, Iran. E-mail: [email protected]; 2CNR, Istituto per la Protezione delle Piante, Via G. Amendola 122/D, Bari 70126, Italy.

NASR ESFAHANI M., HELALAT N. & MOTIEEYAN L. The prospects of non-chemical control of plant-parasitc nematodes in Isfahan, Iran.

The results of studies using farm manures, composts, waste cabbage leaves, and also mulching treatment, and Purpureocillium lilacinum (=Paecilomyces lilacinum) and also the screening of the germ plasm indicating that farm manure including cow dung and chicken manure in integration with polyethylene tarping were the most effective treatments with more than 90% reduction of root-knot and cyst nematodes, followed by the application of these manure alone.The severity of the root-knot nematodes on some of the growing medicinal plants (MP) were assessed. The results indicated that there are species of the MP infected by the root-knot nematode, Meloidogyne javanica, including pot marigold, horehound (Marabium vulgare L.), starflower (Echium amoenum), borage, Klamath weed (Hyperium perforatum L.), absinthium (Artemisia absinthium L.), meadow salsify (Tragopogon pratensis), camomile, garden thyme, greater burdock (Arctium lappa), common sage, Jerusalem Artichoke, Pelargonium, rosemary, milk thistle (Silybium marianum), lemon balm (Melissa officinalis L.), madder (Rubia tinctorum L.), yarrow, common lavender, alehoof (Nepeta hederacea) and celery to varying degrees in Isfahan areas. In Najafabad regions, the infected by M. javanica MP species were common rue (Ruta graveolens L.), Syrian beancaper (Zygophyllum sp.), greater burdock, pot marigold and hempseed (Cannabis sativa L.). In Kashan, common lavender, yarrow, wormweed (Lavandula angustifolia L.), black mulberry (Morus nigra), terracotta gazania (Gazania sp.) and century (Agave sp.) were infected by M. incognita. Most of the medicinal plants in these studies are reported to be new host records for M. javanica and M. incognita. While four species (fennel, spearmint, valerian and yarrow) in Isfahan, and fifteen species (aniseed, blueweed (Echium sp.), cardoon (Cynara drancunculus), dragonhead (Dracocephalum kotschy), wild basil, salad burnet (Sanguisorba minor), hyssop, iris, Klamath weed, lambs ear (Stachys byzanthina), milk thistle, Moldavian balm (Dracocephalum moldavica L.), motherwort (Leonurus cardiaca), oregano and peanut were free from these root-knot nematodes and six species were free from M. incognita. - Plant Protection Research Institute; Center for Agricultural and Natural Resources Research Isfahan, Iran. E-mail: [email protected].

NOWEER E.M.A. Occurrence and morphological features of nematode- antagonistic fungi associated with prevalent nematodes in Abd-elsamad Region Soils (Giza, Egypt).

Occurrence of nematode-antagonistic fungi in Abd-elsamad, Giza, sandy soils which are well-known by its heavy organic manure application were studying by collecting soil samples from fruit orchards, field crop and vegetables during two consecutive years. Five saprophytic fungal species were found, which produce nematode-toxic metabolites in their filtrates namely Aspergillus niger, A. terreus, A. ochrachus, Trichoderma harzianum and T. viride. Seven species of nematode-trapping fungi Arthrobotrys conoides, A. dactyloides, A. oligospora, Dactylaria brochopaga, D. thaumasia var. longa, Dactylella gephyropaga and Stylopaga hadra were isolated from the nematode-positive samples. Four nematode-endoparasitic fungi Catenaria sp. , Cephalosporium balanoides, Haptoglossa heterospora and Harposporium anguillula were isolated from nematode bodies. Verticillium chlamydosporium was isolated from eggmasses of Meloidogyne incognita. All of these fungi were identified, described and photographed. - Plant Pathology Department, Nematology Laboratory, National Research Centre, Dokki, Cairo, Egypt.

ORO V. Heterodera avenae from Serbia and its phylogenetic connections with similar populations.

Cereals have been grown in Serbia and on the Balkan Peninsula since the Neolithic age. Cereals, especially wheat are still a staple food for people from this region. Cyst nematodes associated with cereals were found occasionally and only Heterodera avenae was recorded. Heterodera filipjevi was recently reported in Serbia. Heterodera avenae is one of three main species of the Cereal Cyst Nematode complex. The nematode has been molecularly characterised for the first time. This paper describes phylogenetic relationships between a Serbian population of H. avenae and other similar foreign populations. On the basis of biogeography of host-parasite interaction, in the light of molecular data, the possible centers of oat domestication are also discussed. - Institute for Plant Protection and Environment, Teodora Drajzera 9, 11000 Belgrade, Serbia. E-mail: [email protected].

POUSIS C.1, TROCCOLI A.1, PARK B.Y.2, FANELLI E.1, D'ADDABBO T.1, VERONICO P.1, RADICCI V.1 & DE LUCA F.1 Metagenomic approach for the analysis of nematode diversity and their use as biological indicators.

Nematodes have been used as indicators of soil environment because of their global distribution, ease of extraction and occurrence in trophic groups or functional guilds. Their use as bioindicators has the potential to provide insights into several soil processes. Furthermore, nematodes are sensitive to environmental damage, and variations in their distribution and activity are diagnostic of changes in soil health. Soil nematode community from three selected relatively undisturbed and disturbed sites in Apulia region (Italy) was studied comparatively through morpho-taxonomic and molecular analysis. Nematodes for both analyses were extracted from 100 g sub-samples from composite soil samples collected at each site. Specimens for morphological analysis were fixed in a 2.5% formaldehyde solution and then identified at family and genus level under an optical microscope. The maturity and trophic diversity indices were determined. Total DNA was extracted from the nematode community of each soil subsample and PCR amplification was performed by using the small subunit (18S) of the ribosomal DNA gene, as diagnostic marker. Sequence analysis through BLAST permitted most of them to be classified at genus level and some of them at species level. Maturity and trophic diversity indices were also calculated for genera identified at molecular level. The data obtained revealed that each of the three sampled sites had different level of disturbance. At present more than 300 sequences are available and under characterisation. - 1IPP-CNR, Via Amendola 122/D, 70126 Bari, Italy. E-mail: [email protected]; 2Division of Crop Protection, National Academy of Agricultural Science, Suwon, Republic of Korea. E-mail: [email protected].

PRIDANNIKOV M.V.1, 2 & PETELINA G.G.1 ISTC project on the biodiversity of plant parasitic nematodes in Russia as a basis for multifocal international research of plant parasitic nematodes in Eurasia.

Study of plant-parasitic nematodes in the Russian Research Institute of Phytopathology started in 1955 and continued until 1969. Distribution, biology and pathogenicity of the most important nematodes on vegetables, cereals, potato, onion and garlic were the basic tasks of nematological work. Various methods of chemical, agrotechnical and biological management of nematodes were developed during this time.

A new period of nematological research in RRIP was started by the joint project titled "Discovery of active molecules from natural products for the control of plant parasitic nematodes" together with USDA ARS Nematology Laboratory and financial support by International Science and Technology Center (ISTC) in 2001.

These are the most important achievements of the "Tripartite Alliance" between RRIP, NL USDA ARS and ISTC during the last 12 years:

Twenty regions of Russia (European, Northern-west parts, Caucasia, Ural and Volga region) were investigated for their nematode fauna. Among them the following plant-parasitic nematodes were found: potato cyst nematodes (Globodera sp.), cereal cyst nematodes (Heterodera avenae, H. filipjevi, H. latipons), soybean nematode (H. glycines), sugar beet nematode (H. schachtii), alfalfa nematode (H. medicaginis), root-knot nematodes (Meloidogyne spp.), nematodes virus-vectors (Xhiphinema sp.) etc.

The Collection of plant parasitic nematodes of RRIP was created. More than 600 nematode slides are maintained in the collection and information about those slides is collected in a database. Fourteen species of cyst nematodes, three species of root-knot nematodes are cultured in a greenhouse.

Laboratory of diagnostic of plant pathogenic organisms was founded.

The laboratory renders assistance for various agriculture producers for identification of nematodes by morphological and molecular methods. - 1Russian Research Institute of Phytopathology, Institute Str. 5, RRIP (VNIIF), Bolshie Vyazemy, Moscow region, 143050, Russia; 2Center of Parasitology, A.N. Severtsov Institute of Ecology and Evolution, RAS, Leninsky pr., 33, Moscow, 199071, Russia. E-mail: [email protected].

RASHIDIFARD M.1, SHOKOOHI, E.2, HOSEINIPOUR A.2 & JAMALI S.3 Evaluation of some plant extracts on mortality of second-stage juveniles of citrus nematode.

Citrus nematode, Tylenchulus semipenetrans Cobb 1913, the causal agent of citrus slow decline, is one of the most important nematodes in citrus orchards of the world, and in almost all areas under cultivation of these crops it has caused economic losses. The most effective method for managing the nematode is using chemical nematicides but due to environmental effects, applying non-chemical compounds is increasing worldwide. In this research, the nematicidal properties of four plant extracts including pepper (Capsicum sp.), ginger (Officinale sp.), tobacco (Nicotiana sp.) and fennel (Foeniculum sp.) were evaluated. The experiments carried out in split plot based on completely randomised design, with three replications. The concentration 5% was used for all treatments, with each Petri dish containing 50 individuals of the nematode. The data were recorded at 24, 48 and 72 h after application of the plant extracts. Finally,

the results were analysed using SAS software, and Duncan's multiple range tests were used to compare means. The results indicated that all plant extracts decreased population levels of second-stage juveniles of T. semipenetrans. The highest mortality was observed in tobacco leaf extract (94%, P < 0.05). The lowest mortality was caused by the pepper leaf extract (83%, P < 0.05). - 1Young Researcher Society, Shahid Bahonar University of Kerman, Kerman, Iran; 2Plant Protection Department, College of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran. Email: [email protected]; 3Plant Protection Departments, College of Agricultural Science, University of Guilan, Rasht, Iran.

RENCO M.1 & SASANELLI N.2 In vitro and in vivo effect of chestnut tannin solutions on the potato cyst nematode Globodera pallida pathotype PA2.

An in vitro and an in vivo experiment were carried out to verify the potential effect of chestnut tannin aqueous solutions on the control of the potato cyst nematode Globodera pallida patotype Pa2. In the in vitro hatching test, different tannin concentrations in a geometric scale, from 0.32 to 20.48 g l-1, were used for their effect on hatching of the nematode, whereas in the in vivo pot experiment the soil infested by the potato cyst nematode was treated with three different doses of tannin 100 g m-2; 250 g m-2 and 450 g m-2 applied at potato sowing or at sowing and 2 weeks later to evaluate their effect on nematode control. In both trials tannin treatments were compared to untreated controls. In the in vitro experiment, chestnut tannin solutions significantly suppressed hatching in comparison to untreated control. Pot experiment confirmed results obtained in the hatching test; at all doses of chestnut tannin treatments the number of cyst 100 g-1 soil, number of egg and second-stage juveniles g-1 soil and the reproduction rate r (Pf/Pi) were significantly reduced in comparison to untreated control. Therefore, the use of tannins appears promising for the control of phytoparasitic nematodes in sustainable agriculture. - institute of Parasitology, Slovak Academy of Sciences, Hlinkova 3, 040 01 Kosice, Slovak Republic. E-mail: [email protected]; 2Institute for Plant Protection, C.N.R., Via G. Amendola 122/D, 70126 Bari, Italy. E-mail: [email protected].

ROMANENKO N.D.1, PEREVERTIN K.A.1, POPOV I.O.1, POPOVA E.N.2 & PETRUNYA I.V.1 On the incidence of mixed viral infections and their effects on the yield of potato tubers.

The potato golden cyst nematode, Globodera rostochiensis, was found in tomato and potato plantations of Novoannensky, LLC in the Volgograd region of Russia. Potato virus Y (PVY) and Potato virus X (PVX) were also found on the territory for the first time. PCR analysis revealed potex- and potivirus infection on tomato plants. In addition, the negative effect of a single infection with X, M, and S viruses and mixed infections (X + S, X + S + M, X + S + M + L viruses) was also demonstrated in field-plot tests with potatoes. It was shown that mixed viral infections reduce the yield of potatoes in 2-3 times. As a result of our investigations, we found that Baermann funnel method is not valid for extraction of virus-vector nematodes and decanting and sieving technique should be used in this case. - 1A.N. Severtsov Institute of Ecology and Evolution, RAS, Lenininsky prospekt, 33, Moscow, 119071, Russia; 2Institute of geography, RAS, Staromonetnij per., 29, Moscow, 119017, Russia. E-mail: [email protected].

ROMANENKO N.D.1, PEREVERTIN K.A.1, METLITSKAYA K.V.2, ZAYETS V.G.3, TABOLIN S.B.1, POPOVA E.N.2, POPOV I.O.1 & PETRUNYA I.V.1 A study of biological and economic efficiency of fungal and bacterial antagonists and their possible use in plant protection.

Twelve strains of fungi and bacteria were tested in field, laboratory, and greenhouse experiments and showed high antagonistic activities towards pathogenic nematodes, fungi and viruses. Their biological efficiency varied from 80 to 100% and economic efficiency was 1.3 to 6.0 times greater than untreated controls (data for strawberry and black currant plants). Isolated strains of bacterial and fungal antagonists that possess multifunctional activity include nine strains of bacteria, as follows: four strains of B. thuringiensis, two strains of B. polymyxa, two strains of Pseudomonas fluorescens, one strain of P. aureofaciens, two strains of the fungus Trichoderma viride, and one strain of the predatory fungus Arthrobotrys oligospora. They caused significant reduction in the number of plant-parasitic nematodes and inhibition of fungal, viral and oomycete infections. - 1A.N. Severtsov Institute of Ecology and Evolution, RAS, Leninskii pr., 33, Russia, 119071, Moscow. E-mail: [email protected].

ROMANENKO N.D., TABOLIN S.B. & TITOVA A.S. On the development of environmentally friendly protective measures against pests on strawberry.

As a result of our studies conducted in the end of 20th and the beginning of the 21st centuries, we proposed environmentally-friendly technology for plant protection in strawberries. The technology includes 1) 48.2 C and 48.4°C hot water treatments of plants for 15 min, and 2) treatments of roots with bacterial and fungal bioagents to prevent primary infections during planting. For summer treatments, we recommend the use of fungal and bacterial bioagents

capable of rapid colonisation of plant rhizospheres and suppressing nematode, fungal, and oomycete infections. To control mites we suggest the use of Acarine in combination with oligofurostanoside for strawberry plants under field and greenhouse conditions. - Center of Parasitology, A.N. Severtsov Institute of Ecology and Evolution, Leninsky Pr. 33, Moscow, 119071, Russia. E-mail: [email protected].

RYSS A. The history of Russian nematology: Zoological Institute St. Petersburg.

The history of Russian nematology may be analysed using the example of the Nematode Collection of Zoological Institute RAS (ZIN) with step changes in goals and collection units. 1) Foundation (Ivan N. Filipjev, 1912-1933) of the first Russian lower worms collection of microscopic slides and moist fixations, reference collection of plant-parasitic and entomopathogenic nematodes, development of the first general nematode classification, identification key books of pathogenic nematodes and faunistic compendia; 2) a development of the academic centres in all the national republics of former Soviet Union (initially ZIN branches) with nematode collections and research teams to study biodiversity, morphology, taxonomy, phylogeny; training of plant protection and biocontrol specialists (1930-1980-present); 3) technological revolution (from 1990-present): new types of collection units and research tools: global biodiversity monitoring based on e-technology, digital, molecular and living collections for fundamental research (phylogeny, co-evolution, host-parasite interactions, molecular markers for of pathogenicity, endemic epiphytoties), applied projects (resistant varieties, biological preparations, the use of nematodes as living models and bioindicators in ecomonitoring). Of recent importance is the development of the global net of virtual digitised collections. Results of studies of ZIN collection and its branches were published as papers and in the book series "Fauna of the USSR/Russia" and the "Keys to Fauna of the USSR/Russia". The role of the ZIN Collection in the network of global collection centres continues to grow recently with international cooperation in global biodiversity conservation, environment research, and agricultural crops management. - Zoological Institute RAS, Universitetskaya Nab., 1, St. Petersburg, 199034, Russia. E-mail: [email protected].

RYSS A.1, ANDREEV M.2 & KURBATOVA L.2 Nematodes of Antarctica.

In 2007-2012, 180 samples of mosses and lichens were collected from eight areas (15 stations) around Antarctica. Nematodes were detected in 85% of the moss and 30% of lichen samples; thus indicating that higher moss hydrophily favours the nematode water film habitats. In ZIN collection 58 species of Nematoda (families: Plectidae, Quitsinematidae, Monhysteridae, Aphelenchoididae, Aphelenchidae, Panagrolaimidae, Cephalobidae (with domination of Plectidae and Cephalobidae) and three Tardigrada species were identified. In marine areas nematodes belonging to Axonolaimidae, Comesomatidae, Leptosomatidae, Oncholaimidae, Desmodoridae, Anticomidae, Enoplidae were found.

Fauna impoverishment is demonstrated along a line from South Shetland Islands around the continent. Three succession stages were recognised: i) polytrophic nematodes of the order Plectida on the most poor stone substrates with crustose lichens; ii) omnivores of the order Dorylaimida feeding on big organisms (algae, tissues of mosses and invertebrates in micro-environments with domination of foliose lichens) are added to plectids; iii) in organic substrates with fruticose lichens and mosses, nematodes of the order Rhabditida (including Tylenchina) and Monhysterida with specialised trophic types: myco-, bacterio- and plant-parasitic feeding, are added to nematodes of the previous succession stages. The nematodes of orders Rhabditida and Monhysterida had the highest Maturity Indices for Antarctic communities. The 3D digitised web-atlas of Antarctic Nematode Collection was developed. Fed. Program "World Ocean", Project N 4 "Estimation of ecosystems in areas of the Russian Antarctic Expedition". - 1Zoological Institute RAS, Universitetskaya Nab., 1, St. Petersburg, 199034, Russia; 2Botanical Institute RAS, St. Petersburg, 199034, Russia. E-mail: [email protected].

RYSS A. On the concept of endemic vector-borne transmission infections applied to nematode-caused diseases of plants and animals.

According to the E.N. Pavlovsky concept of endemic vector-borne transmission infections (TI), the main agent of this type of diseases is the pathogen that may successfully be reproduced in phylogenetically unrelated hosts; other important TI conditions are the availability of a pathogen vector, an endemic infection area with the natural reservoir host and the incidental recipient host, which are humans or organisms of economic importance. The concept is well developed for parasitic arthropods, ticks and blood-sucking insects. This concept is proved here to be applicable to nematode-caused diseases as well. In modern nematology, data on the key role of parasitic nematode symbionts in pathogenicity of nematode-bacteria-fungi associations has been accumulated. The TI agent role may be taken by nematodes, as well as their symbionts: bacteria and fungi. Nematodes may serve as TI vectors of agents, insects which transferred nematodes also may be vectors. Natural reservoir host may be mammals, arthropods, or nematodes themselves. The complex TI agent of the plant wilt diseases may include an association of pathogens killing plant host

and after that destroying dead organic material. These are nematode-bacterial-fungi infections, which probably originated from the destroying agent complex in detritus food chain. In the framework of the concept, the diseases caused by nematodes of families Filariidae, Steinernematidae, Aphelenchoididae, are analysed. - Zoological Institute RAS, Universitetskaya Nab., 1, St. Petersburg, 199034, Russia. E-mail: [email protected].

SAIFULLAH A. Low temperature scanning electron microscopic studies on the interaction of Globodera rostochiensis Woll. and Trichoderma harzianum Rifai.

Low temperature scanning electron microscopic (LTSEM) studies revealed that Trichoderma harzianum infected mature potato cyst nematode eggs by penetrating directly the cyst wall or via natural opening of the mouth. Mycelial penetration on cyst wall or egg surface has been seen. The penetration of cyst wall or egg surface was either chemical or mechanical (directly or with appresorium) or both. Freeze fractionation showed the presence of mycelia inside the eggs.

- Department of Plant Pathology, the University of Agriculture, Peshawar-Pakistan. E-mail: [email protected].

SAMALIEV H.1, MARKOVA D.2 & BAICHEVA O.3 Reproduction of root-knot nematodes on eleven common weeds in potato fields in Bulgaria.

Weeds enable plant-parasitic nematodes to survive in the presence or absence of a crop, providing a source of nematode inoculum for the following season. Host suitability studies of 11 weed species commonly found in potato fields in Plovdiv and Pazardjik potato growing region in Bulgaria to two local population of root-knot nematode species (Meloidogyne hapla and M. arenaria) were conducted under greenhouse conditions. Root-galling index, egg mass root g-1, eggs g-1 root and numbers of second-stage juveniles (J2) in soil per pot were recorded at plant harvest. Reproduction factor (Rf = final population/initial population) was calculated to determine the host status for each plant species. The nematodes density in fallow pots was recorded also. Amaranthus retroflexus, Chenopodium album, Cynodon dactilon and Xantium strumarium were not hosts for M. hapla as at the end of the experiment there were no significant differences between the populations in the fallow pots and in these weeds and no egg masses and eggs were observed on these weeds. Cirsium arvense, Echinochloa crus-galli, Lamium amplexicaule and Portulaca oleracea with few egg mass produced per root plant, but maintaining significantly higher population densities in the soil than were recorded in the fallow pots and are considered a poor host. Convolvulus arvensis, Polygonum convolvulus and Solanum nigrum were hosts of M. hapla with Rf of 1.81, 1.61 and 5.50, respectively. Fourteen weeks after inoculation of fallow pots, only 2.93% of the population of M. hapla was still alive. Amaranthus retroflexus, Ch. album, P. oleracea and X. strumarium were hosts of M. arenaria with multiplication rates of 1.5, 1.1, 1.1 and 2.4, respectively. Convolvulus arvensis, C. dactilon, Ech. crus-galli and P. convolvulus were non-hosts as no egg masses and eggs of the target nematode were observed on these weeds. Cirsium arvense, L. amplexicaule and S. nigrum are considered poor hosts with Rf < 1, but C. arvense and L. amplexicaule maintained significantly higher population densities in the soil than were recorded in the fallow pots. After 14 weeks, in fallow pots 3.24% of the population of M. arenaria was still alive.

- 1Agrarian University, 12 Mendeleev St., 4000 Plovdiv, Bulgaria; 2Maritsa Vegetable Crops Research Institute, 4003 Plovdiv, Bulgaria; 3Sofia, Bulgaria. E-mail: [email protected].

SASANELLI N.1, GALLO M.2, CICCARESE A.3, LAQUALE S.1, CICCARESE F.2 & D'ADDABBO T.1 Evaluation of innovative techniques for a sustainable control of root-knot nematodes and Pyrenochaeta lycopersici.

Biocontrol agents and ozone treatments may be effective tools for a sustainable management of root-knot nematodes and associated soil-borne plant pathogens, such as Pyrenochaeta lycopercisi. The effectiveness of soil sub-irrigation treatments with the chitinolytic fungus Aphanocladium album (isolate MX-95) and gaseous ozone was comparatively assessed on tomato in a greenhouse infested with Meloidogyne incognita and P. lycopersici. A 2x107 CFU ml-1 suspension of A. album or 2 ppm gaseous ozone were applied as stand-alone treatment at transplant or split into pre- and post-transplant applications. Tomato yield, root gall index, corky root severity and soil nematode population were recorded at the end of crop cycle. All treatments with the isolate MX-95 of A. album significantly reduced severity of corky root, root gall index and nematode population density in comparison to the untreated control. No statistical differences of corky root severity, root gall index and nematode population density were observed among the different application schedules. Ozone treatments reduced nematode attack from 38% to 57% according to the two different treatments and corky root severity on main roots from 35% to 44% and on secondary roots from 45% to 55% in comparison to the untreated control. Both A. album isolate MX-95 and ozone positively affected tomato yield. -institute for Plant Protection, C.N.R., Via G. Amendola 122/D 70126 Bari, Italy. E-mail: [email protected]; 2Di.S.S.P.A., University "A. Moro", Via G. Amendola 165/A, 70126 Bari, Italy. E-mail:

[email protected]; 3Di.S.A.T., University "A. Moro", Via G. Amendola 165/A, 70126 Bari, Italy. E-mail: [email protected].

SCHUKOVSKAYA A.G.1, TKACHENKO O.B.1 & SHESTEPEROV A.A.2 Reproduction of mycotrophic nematodes Aphelenchus avenae, Paraphelenchus tritici, Aphelenchoides saprophillus on fungal mycelium Microdochium nivale (Fr.) Samuels & I.C. Hallet, a cause of pink snow mold on winter wheat.

The effect of three temperature regimes (+5°C, 15°C and 27°C) on the growth, development and nutrition of mycohelminths Aphelenchoides saprophillus, Paraphelenchus tritici and Aphelenchus avenae on fungus Microdochium nivale was shown. Temperature +5°C was the most favorable for reproduction of these three nematode species and the total number of nematodes grown at +5°C was about 1.5 times higher than at 15°C and 27°C. Nematodes were able to destroy the entire mycelium during 75 days, whereas at +15°C and +27°C from 20 to 25% of the mycelium remained on the surface of growing medium. Sex and age composition of nematodes at these temperatures vary considerably. The mycohelminth A. saprophillus was the most aggressive towards a low-temperature fungus M. nivale. It was able not only to destroy the entire mycelium, but also to achieve high population numbers. This could be explained by a peculiar property of the mycohelminth: its ability to feed and grow at low temperatures. Conditions favourable for the growth and development of mycohelminths, created during the cultivation (optimal temperature, a complete food base), may promote the mass inoculum of these potential biological agents for low-temperature crop pathogens control. - 1Main botanical Gardens n.a. N.V. Tsitsin RAS, Moscow 127276, Russia. E-mail: [email protected]; 2Russian State Agrarian External University, Balashicha, 143900, Russia. E-mail: [email protected].

SEDIQI, E.1, SHOKOOHI, E.2 & KARIMI, J.3 Molecular characterisation of new isolate of Heterorhabditis bactriophora Poinar, 1975 based on sequence of the ITS rDNA region, from South East Iran.

Entomopathogenic nematodes (EPNs) are among the important biological control agents. During a survey on this group of nematodes, a new isolate belonged to the Heterorhabditis genus was recovered from Lalezar region, Kerman province (Iran). After initial characterisation, the ITS rDNA gene was amplified, sequenced and used for molecular characterisation. The analyses showed that the new Heterorhabditis isolate belongs to "bacteriophora" species group. The phylogenetic study based on Maximum Likelihood method indicated that the new Iranian isolate is in a clade among some other populations of H. bacteriophora. Pairwase genetic distance analysis revealed no genetic variation between Iranian population and the other populations of H. bacteriophora. This is the first record of H. bacteriophora from south of Iran. - 1Young Researchers Society, Shahid Bahonar University of Kerman, Kerman, Iran; 2Department of Plant Protection, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran. E-mail: [email protected]; 3Department of Plant Protection, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.

SHOSHIN A.V.1 & SHOSHINA E.A.2 Morphology and evolution of the lip region of nematodes of the family Tobrilidae.

Lip region morphology is a feature that has not been used in the description of tobrilids due to complexity of observation. The present study is focused on the description of the lip region structure of thirteen tobrilid species. Observed variations of the structure can be divided according to the type of symmetry: triradial and hexactinal. The most primitive triradial symmetry of the tobrilid lip region is characteristic for Asperotobrilus aculeatus. Hexactinal symmetry with respect to large, fully closing mouth opening, symmetrically arranged lips, may be considered the basic state of tobrilini and neotobrilini lip area. In these cases, the structure of the mouth parts do not show any, or only insignificant, features of specialisation. Common specialisation features of the oral apparatus were revealed in tobrilids with a known type of feeding. Tobrilids feeding on diatoms are characterised by the following features: stoma conical, folded, with marked sub-bucal mounds, stoma armament reduced or absent, lips are spliced into a ring. Macro-predators have the following features: stoma has sclerotized walls and is heavily armed, mouth opening wide, lips reduced. To understand where lip structure characteristic for tobrilids originated, the morphology of its (presumably) sister group, belonging to Tripylidae family, should be considered. The representatives of the latter have triradial symmetry of the lip region and its arrangement is similar to one of the most primitive tobrilid, the genus Asperotobrilus. - Zoological Institute of the Russian Academy of Sciences, Universitetskaya nab.1, St. Petersburg, 198342, Russia. E-mail: [email protected]; 2St. Petersburg State University, Universitetskaya nab.7/9; St. Petersburg, 198342, Russia. Email: [email protected].

SHOSHIN A.V.1 & SHOSHINA E.A.2 Morphology of supplement organs of nematodes of Tobrilidae family.

The structure of the supplement organs, as well as the structure of the stoma are basic characteristics of the family Tobrilidae. Four main types of the tobrilid supplements can be distinguished on the basis of their structure. Type I -typical for Tobrilus, Lamuania and Semitobrilus. Supplements are small, external part of the supplement is slightly protruded. There are two variations of the type I supplement structure - "amabilis" and "gracilis". Type II - typical for some Eutobrilus (E. peregrinator, E. prodigiosus, E. strenuus and E. nothus). These supplements are very similar to type I supplements, but have high protruding shoulders with numerous microspikes. Bulbus is situated on the base of an ampulla. Type III - typical for some Eutobrilus (tribe Tobrilini) - E. graciliformes, E. papilicaudatus, E. differtus and Mesotobrilus (tribe Paratrilobini). Supplements have a well-defined cap; a bulb is situated on the base of an ampulla. Type IV - observed in majority of Eutobrilus, Paratrilobus, Brevitobrilus, Neotobrilus. This is the most complicated supplement type with the mobile cap and the upper bulbus.The process of development of nematode sensory organs from unspecialised setae may be illustrated by the evolution of tobrilid supplements. - 1Zoological Institute of the Russian Academy of Sciences, Universitetskaya nab.1, St. Petersburg, 198342, Russia. E-mail: [email protected]; 2St. Petersburg State University, Universitetskaya nab.7/9; St. Petersburg, 198342, Russia. Email: [email protected].

SLOS D.1, ENSAFI P.1, CLAEYS M.1, YUSHIN V.V.2 & BERT W.1 Ultrastructure of sperm development in the peanut-parasitic nematode Ditylenchus n. sp.

The spermatogenesis of the pea-nut parasitic nematode Ditylenchus n. sp. has been studied with transmission electron microscopy. Spermatozoa of Ditylenchus n. sp. represent an aberrant type of male sperm because of the absence of an axoneme and acrosome, a characteristic shared with other nematodes. The spermatogonia consist of cells with a large centrally located nucleus surrounded by a small amount of cytoplasm. The sperm development includes the formation of complexes of fibrous bodies (FB) and membranous organelles (MO) which appear in the spermatocytes. These complexes start to dissociate in separated MO and FB in the spermatids. Immature spermatozoa are unpolarised cells with a centrally located nucleus surrounded by spherical fibrous bodies and MO located at the periphery. The spermatheca contains chains of mature spermatozoa consisting of amoeboid bipolar cells subdivided in a pseudopod devoid of organelles and a main cell body. The main cell body consists of a centrally located nucleus lacking a nuclear envelope, many mitochondria and MO. The MO are connected to the plasmalemma and open to the intercellular exterior via a pore. The general pattern of spermatozoon is recognised as 'rhabditid', a pattern that is rather conserved throughout the order of the Rhabditida. The relevance of our data to study on nematode spermatogenesis in a phylogenetic framework will be discussed. - hematology Unit, Department of Biology, Ghent University, K.L. Ledeganckstraat 35, 9000 Ghent, Belgium. E-mail: [email protected]; 2A.V. Zhirmunsky Institute of Marine Biology, FEB RAS, Vladivostok, Russia.

SPIRIDONOV S.E.1, ODOYEVSKAYA I.M.1, 2, BUKINA L.A.3 & CHILJUTA N.V.2 Discrimination of Trichinella species from Russian Arctic with Cob mtDNA sequences.

Cultures of Trichinella spp. were established from the biopsy material collected in Russian Arctic through the digestion of muscular tissue samples and inoculation of laboratory animals with obtained juveniles. Samples were obtained from polar bear, seals, wolverines and also stray cat and arctic foxes from a fur farm in the Russian Arctic. The partial sequences of Cob mtDNA were obtained for these Trichinella samples using primers Tricob F1 CAA TCC ATT AGG TAC ACA CTC AC and Tricob R3 TAA GTA AGA TTT CAA TGG CG (Rosenthal et al, 2008). For obtained sequences approx. 985 bp long alignment was combined and used for phylogenetic analysis. Only two sequences of the studied cultures belong to Trichinella spiralis. The majority of studied trichinellids clustered with T. nativa and Trichinella sp. T6. Cultures from Russian Arctic differ from sequences of these two species deposited in NCBI GenBank in 2-14 bp. (Note: Trichinella sp. T6 differs in obtained Cob mtDNA sequence from T. nativa in single nucleotide only.) The culture of Trichinella sp. established from muscular tissue sample of stray cat (shot on the fur farm in Chukotka peninslula) demonstrated numerous differences with other Trichinella sequences: 30-35 bp difference with T. nativa and Trichinella sp. T6 complex and 47-48 bp with T. spiralis. In conclusion, an analysis of nucletotide sequences of Cob mtDNA does not reveal significant differences between genotypes T. nativa and Trichinella sp. T6, considered now as independent species, but demonstrated high level of diversity of Trichinella cultures from the Russian Arctic. - 1A.N. Severtsov Institute of ecology and evolution RAS, Leninskii pr., 33, Moscow, 199071; Russia. E-mail: [email protected]; 2Skrjabin's All-Russian Institute of helminthology, Moscow. 3Vjatka State Agricultural Academy, Kirov.

SUSHCHUK A.A. & MATVEEVA E.M. Soil nematode communities of northern insular biocenoses.

Fauna and community structure of soil nematodes of islands situated on Ladoga and Onega Lakes (Republic of Karelia) were studied. Totally, there were 26 islands where 35 different types of habitats were identified: 14 forest (pine, spruce forest, lime, mixed), 15 meadow, 3 transformed and 3 rocky biocenoses. High diversity (79 genera) nematode fauna was characteristic to insular biocenoses. Nematode taxa uncommon for Karelia were identified (Prodorylaimium, Coomansus, Discolaimus, Cephalenchus, Gracilancea and others). Bacterial-feeders were the predominating trophic group. Fungal-feeders were subdominants in forest habitats, and fungal-feeders either nematodes associated with plants in meadows. The proportion of phytotrophs was usually 15% in forests and 21.7% in meadows. However, the differences between island meadows of two lakes were revealed. The proportion of phytotrophs in nematode fauna was higher in the southern region of Ladoga Lake compared to islands of Onega Lake (25.8% vs 13.5%). Soil nematode community structure and ecological indices were similar for the forest and meadow insular biocenoses. It seems that this pattern is linked with the afforestation of meadows, especially in fragmentary lands. Organic matter decomposition by bacteria occurred (CI = 25.6 in meadows, 37.3 in forests). Soil food web was characterised by a high level of complexity and structured condition (SI = 70.2 in meadows, 74.7 in forests). The study was supported by The Ministry of Education and Science of Russian Federation, project № 8101. - Institute of Biology of Karelian Research Centre of RAS, Pushkinkaya St., 11, Petrozavodsk, 185910, Russia. E-mail: [email protected].

TABOLIN S. On the nematode fauna in soils from parks of Moscow.

During the autumn of 2011 and the autumn of 2012, 130 soil samples were collected from various locations in the Neskuchny garden and the Pokrovskoye- Streshnevo park. Despite the difference of flora in the parks, nematode species composition was similar. Nematode trophic groups were represented by bacterial feeders, fungal feeders, plant parasites, predators, and omnivores. Ring nematodes (Mesocriconema xenoplax, Criconema annuliferum), virus-vector nematodes (Longidorus elongatus, Trichodorus primitivus, Paratrichodorus pachydermus), reniform nematode (Rotylenchus robustus), and spiral nematodes (Helicotylenchus digonicus, H. vulgaris) were predominant plant-parasitic species in the parks. The most widespread predatory nematodes were Aporcelaimus pachydermus, Anatonchus tridentatus, Mononchus truncatus, M.aquaticus, Clarkus papillatus, Coomansus parvus, Mylonchulus brachyuris, M. sigmaturus, M. sexcristatus, Prionchulus muscorum, Tigronchoides ginglymodontus, Discolaimus major and Tripyla affinis. During the survey, plant-parasitic nematode Hemicycliophora macristhmus and predatory nematode Tigronchoides ginglymodontus were registered for the first time in Russia. - A.N. Severtsov Institute of Ecology and Evolution, RAS, Leninsky prospekt 33, Moscow, 119071, Russia. E-mail: [email protected].

TCHESUNOV A.V. Peculiarities of fine morphology of Desmoscolecida using the example of Tricoma albimaris.

Species of the order Desmoscolecida are characterised by unusual appearance with prominent cuticular main ring covered with rough concretions. Most authors considered the concretions as a product of secretory activity. Riemann & Riemann (2010) basing on observations on behaviour of live desmoscolecoids conclude that concretions are mineral particle accumulations. The goal of this study is evaluation of both hypotheses of the cuticle structure as well as obtaining new data on other desmoscolecoid strucrures. The main subject is Tricoma albimaris Decraemer & Tchesunov, 1996 sampled in the sublittoral zone of the White Sea, Northern Russia, and an additional subject is Quadricoma sp. from Pacific deep sea. TEM and SEM microscopy were used, together with SEM detection of element composition. Covering over the main rings appears as disorderly accumulations of irregular amorphous platelets or granules of various shape and size. No glands or any cells with secretory activity were revealed in the subcuticular epidermis. Analysis of element composition reveals clear peaks of silicium, aluminium, and iron that indicate the concretions are mineral accumulations resembling clay. The hypothesis of Riemann & Riemann (2010) is thus supported. It was also found that large vesicular amphids are covered with thin cuticle, which is similar to the body cuticle and bears concretions. So-called ocelli known in most desmoscolecoid species including deep sea species are ovoid bodies with concentric striation and without evident sensory structures. The project is by grant RFBR 12-0400781. - Department of Invertebrate Zoology, Faculty of Biology, Lomonosov Moscow State University, Moscow, 119991, Russia. E-mail: [email protected].

VAISH S.S. & SINGH K.P. Catenaria anguillulae Sorokin: its potential and prospect for biological control of plant-parasitic nematodes.

Use of biological control agents for management of plant diseases offers several benefits over the use of chemicals, mainly ecological safety and safe and sustainable better human life. Nature has provided several predaceous fungi to

regulate population of nematodes. Among predaceous fungi, Catenaria anguillulae seems to play a key role in modulating population of nematodes in soil as it has several attributes of a good biological control agent viz., fast multiplication at high rate, epidemic-causing ability, facultative endoparasitic nature as it colonises plant debris as well as susceptible nematodes with varying degree of virulence. Moreover, C. anguillulae is widely distributed and has good compatibility with agrochemical and fertilisers. Therefore, it is suitable for the integrated management of plant diseases caused by nematodes. Catenaria anguillulae is a zoosporic nematophagous facultative endoparasitic blastocladian fungus producing sporangia in chain. This fungus can be used as the best educational tool to demonstrate biological control in class room without any chemical input in water only. Additionally, several techniques for its selective isolation, semi-quantification, rapid virulence test, purification have been developed and substrates for its mass culture are known. However, there is a need to screen cheaper and easily locally available substrates. This preliminary work clearly reflects great potential in C. anguillulae as an appropriate biological control agent. However, the extensive work on its nutritional requirement, refinement of its mass culture technique including formulation development, ecological aspects and performance evaluation against agriculturally important plant-parasitic nematodes is still required for its field application. - Department of Mycology and Plant Pathology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi- 221 005, India. E-mail:[email protected].

WEN Y., MEYER S.L.F., MASLER E.P., LIAO J. & CHITWOOD D.J. Toxicity of alkaloids from the Chinese plum yew (Cephalotaxus fortunei) to Meloidogyne incognita, Panagrellus redivivus, and Bursaphelenchus xylophilus.

Cephalotaxus species are known to produce unidentified compounds toxic to nematodes. Therefore, a bioassay-guided chemical fractionation process was utilised to identify the compounds responsible for inhibition of reproduction of the pinewood nematode, Bursaphelenchus xylophilus. Inhibitory activity primarily occurred in a fraction containing a mixture of alkaloids; further purification revealed that the alkaloid drupacine was the major inhibitory component. The ED50 of drupacine for inhibition of B. xylophilus reproduction was 27.1 ^g ml-1. In a Meloidogyne incognita direct contact assay with second-stage juveniles (J2), the ED50 was 76.3 ^g ml-1, and immobile J2 did not regain motility when transferred to water. Crude alkaloid extract exhibited greater toxicity to eggs (as indicated by hatching experiments) than J2 of M. incognita. In a stylet thrusting/motility bioassay, nematode activity was inhibited fourfold by 20 ^g ml-1 of crude alkaloid extract. Drupacine at 0.3 mg ml-1 inhibited protease activity in Panagrellus redivivus extracts more than crude alkaloid extract applied at the same concentration. Although protease activity in extracts of M. incognita was lower than that in P. redivivus extracts, it was also significantly inhibited by drupacine. In greenhouse experiments on Capsicum annuum, application of 0.5 mg ml-1 crude alkaloid extract to soil infested with M. incognita did not significantly reduce shoot length or weight, but the number of eggs and J2 per root system decreased by 69% and 73%, respectively. This research provides the first demonstration of nematotoxicity of crude Cephalotaxus alkaloids and the purified component drupacine. Moreover, crude alkaloids extract effectively suppressed root-knot nematode egg and J2 production on plant roots. Although the specific mode of action of drupacine and other cephalotaxine alkaloids against nematodes remains unknown, this is the first demonstration in any nematode that a plant alkaloid inhibits proteolytic activity. - USDA ARS Nematology Laboratory, 10300 Baltimore Ave., BARC-West, Bldg 10A, Beltsville, MD, 20705, USA.

WESTERDAHL B.B. Population cycling of plant parasitic nematodes in perennial cropping systems.

Nematode population cycling in soil and roots is driven by biotic and abiotic factors. In California, USA, nematode population cycling and associated abiotic factors have been monitored over several growing seasons, for dagger nematode (Xiphinema index) on grapes; lesion nematode (Pratylenchus penetrans) on Easter lilies; ring nematode (Mesocriconema xenoplax) on prunes; lesion (P. vulnus) and ring nematode on walnuts; and for Anguina pacificae and other nematodes on turfgrass. Yearly trends vary by cropping system, but are reasonably consistent for a particular crop. Results have made it possible to improve recommendations for methods to sample vineyards, orchards and golf courses; improved our ability to interpret nematode samples obtained at different times of the year; and initiated additional research on timing of management practices. - Department of Entomology and Nematology, University of California, One Shields Ave., Davis, CA 95616 USA. E-mail: [email protected].

YUSHIN V.V.1, 2, CLAEYS M.3 & BERT W.3 Major sperm protein (MSP) in the nematode spermatozoa.

The nematode spermatozoa represent a highly modified (aberrant) type of male gametes which lack a flagellum but develop ancient crawling type of movement on the basis of the cytoskeletal component known as "major sperm protein" or MSP (nematode specific small protein of 126 amino acids with a molecular weight of 14 kDa). Despite the apparent novelty of MSP and sperm motility, the system can offer insights into the canonical, actin-based mechanism of cell

movement. The MSP is also known as an important hormone triggering oocyte maturation and stimulating the oviduct wall contraction to bring oocytes into position for fertilisation. MSP seems to be restricted to nematodes, and MSP-based sperm motility appears to be conserved among nematodes. Initially MSP has been identified in Caenorhabditis elegans; then the MSP genes were sequenced in many distant nematode taxa. However, direct evidences of MSP localisation and fate in spermatogenic cells are rare. New data on MSP presence and localisation illustrate form and function of MSP in very diverse spermatozoa of nematodes. MSP begins to assemble in spermatocytes as paracrystalline fibrous bodies (FB) associated with the membranous organelles (MO); assembling continues through the spermatocyte stage; the FB-MO complexes disassemble in the spermatids and upon activation, MSP reassembles into filamentous fibers in the pseudopod. The immunocytochemical observations show presence of MSP in spermatozoa of very distant taxa of nematodes. (Support: RFBR 11-04-00368; FEB RAS 12-III-A-06-098; RF 2010-220-01-180). -1A.V. Zhirmunsky Institute of Marine Biology, FB RAS, Vladivostok, Russia. E-mail: [email protected]; 2Far East Federal University, Vladivostok, Russia; 3Nematology Unit, Department of Biology, Ghent University, Belgium.

ZHILINA T.M. & SHEVCHENKO V.L. Soil nematodes in pine forests of natural reserved territories of Chernigov Region.

Soil nematode fauna in pine forests of natural reserved territories of Chernigov region was studied. Thirty species of nematodes belonging to four orders were found. Rhabditida and Tylenchida dominated in number. Nematode-saprobionts prevailed in pine forests of region. - Chernigov National Pedagogical University after T.G. Shevchenko, Chernigov, Polubotka St., 53, Ukrane, 14013. E-mail [email protected].

ZHILOKOV A. & KOZHEVNIKOV K. Electronic accounting of pathogen biological agent collections (PBA).

The issues, which arise at the junction of various spheres of science and activity, have always been the most interesting and frequently complex from the point of view of finding the optimal and elegant solution. One of such issues is building Electronic Systems for tracking and control of the Pathogenic Biological Agents (PBA) stored in collections of scientific research institutes and laboratories. The system should provide the required level of security and reliability of PBA data storage, which is confidential and should correspond to the existing rules on record keeping of biological materials. It has to be simple and convenient to use and to provide additional advantages for use of the information compared with the paper records. Pathogen Asset Control System (PACS) is an electronic system for accounting and control of pathogenic materials, which are kept in laboratories, with many features for inventory management and reporting, and provides high level of security and reliability of the information storage. The user interface of the application is modern, simple and convenient to use, and facilitates easy learning to start working quickly with PACS. The system was built from scratch based on subject matter expertise provided by laboratory professionals from many scientific organizations. Currently available PACS version 4 offers integration with Radiofrequency Identification (RFID) technology. Version 5 development is ongoing and release is planned for Autumn 2013. - PACS Analyst, Federal Services Division, Black & Veatch, 16 Marksistskaya St., Moscow, 109147, Russia. E-mail: [email protected], [email protected].

ZINOVIEVA S.V. Plant resistance to parasitic nematodes: molecular genetic and biochemical aspects.

Sedentary plant-parasitic nematodes establish long-term relationships with their hosts. Gene expression studies of several plant-nematode interactions showed that different defence-related genes are upregulated upon infection of both susceptible and resistant plants, including genes encoding peroxidase, chitinase, lipoxygenase, extensin, and proteinase inhibitors. Expression of these defence-related genes in both the compatible and the incompatible interaction suggests their role in basal resistance. The main components of plant immunity to plant-parasitic nematodes are specific membrane and cytoplasmic receptors. These receptors possess conserved leucine - rich repeats (LRR) patterns. Membrane-bound pattern recognition receptors play the major role in the non-specific immune response by triggering necessary reactions in order to fight the pathogen. However the defence response is strong and quick enough to prevent successful nematode infection only in the presence of a functional R protein that can recognize the appropriate AVR protein from the nematode. Nematode resistance genes are members of the LZ-NBS-LRR family. The LRR region plays a role in the signalling process that leads to plant cell death (typical for the hypersensitive response) and the N-terminal part of the protein controls this cell death. The Hspro1 gene (from wild beet), which confers resistance to Heterodera schachtii is different from any other resistance gene cloned from plants. The gene encodes a relatively small protein that does not contain a LRR. - A.N. Severtsov Institute of Ecology and Evolution, RAS, Leninski prospect 33, Moscow, 119071, Russia. E-mail: [email protected].

ZINOVIEVA S.V.1, UDALOVA ZH.V.1 & GERASIMOVA N.G.2 Role of jasmonic and salicylic acids in the resistance of tomato to root-knot nematode Meloidogyne incognita.

Salicylic (SA) and jasmonic (JA) acids are the best known mediators of signal systems in plants. Participation and character of interactions between SA- and JA-signals under conditions of the induced and genetic resistance of plants to nematodes were studied. Model system of a tomato (Solanum lycopersicum) and a root-knot nematode Meloidogyne incognita was employed. This study demonstrated that application of JA and SA to tomato foliage induced systemic effects that suppressed root-knot nematode infestation, inhibition of nematode reproduction, and also increased activity of LOX and PAL, and the enzymes of biosynthesis of JA and SA. JA treatment did not inhibit M. incognita-mediated resistance, which suggests a lack of signalling conflicts between these two forms of defense. - 1A.N. Severtsov Institute of Ecology and Evolution, RAS, Leninsky pr., 33, Moscow, 119071, Russia. E-mail: [email protected]; 2Bach Institute of Biochemistry RAS, Leninsky pr., 33, Moscow, Russia, 119071.

ZOGRAF J.K. & YAKOVLEV K.V. Spermatogenesis and localisation of major sperm protein (MSP) in the free-living nematode Panagrellus redivivus (Rhabditida, Panagrolaimidae).

Nematode spermatozoa are nonflagelated cells moving by the means of pseudopodium. Molecular composition of the nematode sperm cells has been studied on the example of free-living soil nematode Caenorhabditis elegans and parasitic nematode Ascaris suum. It was shown that major sperm protein (MSP) comprises 15% of total proteins in the sperm cell and it was not found in any other tissue of nematodes. MSP is able to form filaments that are playing role in the cellular locomotion. Though the structure and function of MSP has been accurately studied, the presence of the MSP in the nematode spermatozoa has been shown on the little number of species. Localization of the MSP in germ cells of the Panagrellus redivivus has been studied using confocal microscopy. During spermatogenesis, MSP is first found in the spermatocytes II in granular form and number of granules increasing during cells growth. In spermatids MSP still present in the granular structure and fills almost all cytoplasm. In spermatozoa from the vesicula seminale granules with MSP dissociate and fibers of MSP fills cell cytoplasm. Activated spermatozoa from the female reproductive system are found in two conditions: i) free spermatozoa in the uterus lumen; ii) spermatozoa attached to the uterine wall. In the first case spermatozoa are polarized cells with anterior pseudopodia and posterior main cell body. In this case, fibers of MSP filled pseudopodium cytoplasm. Spermatozoa attached to the uterine wall are not polarized cells and MSP evenly distributed in the cell cytoplasm. (Support: RFBR 11-04-00368; FEB RAS 12-III-A-06-098; FEB RAS 13-III-B-06-013; RF 2010-220-01-180). - A.V. Zhirmunski Institute of Marine Biology FEB RAS, Vladivostok, 690059, Russia. E-mail: [email protected].

ZULLINI A. Is a biogeography of freshwater nematodes possible?

An attempt was made to detect some possible biogeographic patterns in the distribution of freshwater nematodes. The literature concerning 14 geographic areas (in Eurasia, Africa and America) and 8 well studied lakes, was examined, i.e. many papers published from 1905 to 2011. After some taxonomic corrections, a total of 1030 nominal species were listed: the richest area was Germany + Austria (301 valid freshwater nematode species), whereas the richest lake was Balaton (143 species). Notwithstanding the ubiquity of most freshwater nematodes, some distributional patterns emerge from a multidimensional analysis: e.g., the nematode communities of the European regions are more similar inter se in comparison with the other continents. Examining the lakes only, Baikal nematodes appear more different from the other lacustrine nematodes from other parts of the world. It is possible to conclude: (1) that freshwater nematodes communities show some difference at the continental level only, and (2) that the freshwater nematode biota is divided into two parts with the Lake Baikal community on one side, and the rest of the world community on the other. -Department of biotechnology & biosciences, University of Milano-Bicocca, Italy; E-mail: [email protected].