The effect of deregulated JAK/STAT signaling on the structure of the fruit fly's respiratory epithelium Christine Fmk1 •, Khnberty Kallsen2, Ruben PrangeMarcus Thiedmamv, Hotger Heine2 and Thomas Roeder' 'Molecular Physiology. Zoological Institute, Kiel University (Germany) annate itrwnunity. Asthma & Allergy Research Center Borstel (Germany) Abstract The JAK/STAT signaling pathway is an evolutionary highly conserved pathway that is involved in developmental processes like cell proliferation, differentiation, cell migration and apoptosis. Moreover, it ptiys an important role in the development of the innate immune system.__ ^4^,^^^ The canonical pathway is activated by mainly two groups of ligands, chemokines and growth factors. After the activation of the receptor the intracellular Janus Kinase (JAK) leads to tyrosine phosphorylathn and to STAT activation. We are interested in the role of JAK/STAT signaling in the fly's airway epithelium under different stress conditions like oxidative stress, cigarette#moke and hypoxia. Furthermore, we wanted to know if the secretion of the ligands of the ^■aij^djaniilvjtf^fis regional specific in the tracheal compartments atd if a constitutive overexpression of upds leads to a structurallycnMge of the airways epithelial cells. • 0 4 With our results, we were able to show that there is a time dependent expression of two of the three unpaired ligands, upd2 and upd3 in oxygen undersupplied wildtype flies. By enhancing expression of different components of the JAK/STAT pathway we could observe structural changes in the dorsal trunks of the fly's airway mirrored by epithelial thickening and meta- as well as hyperplasia or even lethal effects by constitutive activation of the Dome-receptor in late larval stages. ne expression analysis ^ Cytokine expression Changes in epithelial structure Hyperplasia X ✓ ✓ ✓ Hypoxic conditions [5% OJ lead to time dependent expression of JAK/STAT activating genes. 3. 3£ 10KH Ectopic ac f I upd3Hil / f B. 200 n i—■ fJT J PmMMUiMf'l ^^^H f-i m i*o 4 m ■ P'»oU"FM«>lCkloopor,JAK/$TAT.i9ntlins y opic activation of man/ jAK/STAT components lead lo a compact increase ol the epithalial thickness. ^^/^S^^^^S^^i ^^^^ Ectopic activation of man» JAK/STAT components lead to a compact increase of nuclei number & nuclei size. C0„.,l,uU,.dom...,p,...lon,.u-. to Intens...... early daaln {Li larvae) Crosstalk with other Pathways iL iL iL, 's SptCiMl 10 tii ,11.1 LWMM'W P05 Drosophila melanogasteris a model for the study of Bacillus cereus pathogenicity Zaynoun Attieh1'2, Agněs Rejasse2, Christina Nielsen-Leroux2, Mireille Kallassy1, Vincent Sanchis2, Laure El Chamy1 1- Unite de recherche Environnement, Génomique et Protéomique, Laboratoire de Génétique de la drosophile et virulence microbienne. Universitě Saint-Joseph, Beirut Lebanon 2- Micalis Institute, INRA, AgroParisTech, Universitě Paris-Saclay, 78350 Jouy-en-Josas, France Introduction Bacillus cereus sensu lato Drosophila immune response 3Ö1 Bacillus ccrcus Baciilus o totoxicus Food bone disease Bacillus anthracis Anthrax' Bacillus thuringiensis "Errttwmopathorjen" Pathogenic bacteria manipulate the host immune responses through , the activity of virulence genes. The identification of these genes is of | particular interest since it is essential for the onset of new therapeutic strategies for infectious diseases. The Bacillus cereus group includes eight species of Gram-positive sporulating bacteria. These bacilli share a highly similar genetic background with particular virulence genes enabling them to colonize different hosts. Taking advantage of Drosophitís powerful genetic tools and its long studied immune system, we use it as a model organism for the study of B. cereus pathogenicity. Using a septic injury infection model, we showed that 8. cereus is highly lethal to the flies. We then screened a B. cereus mutants library to isolate and characterize non-virulent mutant bacterial clones. In a complementary approach, we set up a Drosophila oral infection model to investigate the infectious process of food-borne B. cereus toxi-infections. Systemic Humoral response Production of antimicrobial peptides (AMPs) I <><5^0<5ooOc>0( I Cellular response j Phagocytosis Epithelial defenses AMPs and Reactive Oxygen Species (ROS) Identification of bacterial virulence genes Screening of a B. cereus mutants library in a Drosophila septic injury infection model Research interests and work strategy (l^) Screening by survival assay Characterization of food bo infections Screening of natural 8. cereus strains in a J Drosophila oral infection model Selection of non-virulent bacterial clones (js^ Phenotypic characterization of the non virulent mutants Characterization of inducible immune response Results I- Characterization of 5. cereus pathogenicity in a septic injury infection model Set-up of the infection model and screening strategy B. cereusn resistant to the Drosophila systemic humoral response Selection of bacterial mutant clones with attenuated virulence B. cereusn highly lethal to Drosophila Wt (PBS) -*■ wt (Sc - pellet) -*- wt (Be 00*00=1) Wt(Bc ODsoo^) i * 6 g 10 12 14 16 li lime (Hours) Survival of wld-type fl.es (wt) to in .nfectior. with wild-type 8. cereus [8c] at d i Her ent concentrations Phenotypic characterization of bacterial mutant clones All selected mutant clones have a reduced biofilm forming c Survival of wild-type (Wt) and Relish i -iuuki 1 - -___ i II- Isolation of a virulent lone A Clone B Clone C Bacterial motility o Statistical test studeníš cereus strain for the study of foodborne i LB medium U"*p*0 00L) } 90 1?0 1W> 180 ?10 2*0 270 tOO IK Time (mlnutet) Bacterial growth on LS medium -*- Wt(ftr) f ■ del (at j -•- Wt (Clone A) Rel(CloneA) Time (Hours) Survival of wild-type (Wt) and Relish (Rei) mutants fr« to an infection with wild-type a cereus {act and the mutants donei toxi-infections in Drosophila '^(QaytJ Contact ood poisoning an(J dlniM| fl mmla %tnin% Wh[|e Vpe strain .s not pathogenic, strain 3 is lethal to files when ingested via contammated (ood resn.,,r« Conclusions Using a septic injury infection model in Drosophila, we isolated three non virulent mutant of B. cereus, each affected in a different gene. The phenotypic characterization of the mutants indicate that the three selected clones are affected in their biofilm formation capacity and motility. Our results also put forward the resistance to the systemic humoral AMP-dependent response as a prominant aspect of 8. cereus virulence mechanisms in Drosophila Moreover, in an oral infection model, we isolated a virulent B. cereus strain that is lethal to the flies. This model will be used for the investigation of the infectious process of food-borne A cereuslQxi-mfect\ons in Drosophila.___ Zaynoun ATTIEH(uyriounattl(,h Laur« f| CHAMY Or 11» """Wnet usj.edu.lb «nt SANCHIS BORJA. Or I «"«nt.unchit.barlaein«.lr) Acknowledgment TRIM USJHi 4*p ä m mmSßs Using Drosophila to understand the genetic basis of obesity Neha Agrawal1,1. Sadaf Farooqi2 and Andrea H. Brand1 Ag 'Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge United Kingdom ZW % J institute of Metabolic Science, University of Cambridge, Addenbrooke's Hospital, Cambridge, United Kingdom. BS Obesity is one of the greatest public health challenges facing the world today with the numbers of those affected continuing to rise at an alarming rate. Obesity ultimately represents an imbalance in the body's capacity to maintain energy homeostasis and is influenced by complex interactions between genetic and environmental factors. The Genetics of Obesity Study at the Institute of Metabolic Science, University of Cambridge has done pioneering work in identifying genetic factors underlying human obesity. We are now developing Drosophila models to identify and examine obesity causing genes and generate patient-specific obesity models, in collaboration with this study. This research will thus help identify novel genes and underlying cellular and molecular mechanisms involved in modulating energy homeostasis which will ultimately contribute to strategies for the prevention and treatment of obesity. Human obesity is influenced by complex interactions between genetic and environmental influences DNA H.im JOCJ. Í0O7. uoww^.xrn Drosophila melanogasterwiW be used as a model system to examine novel obesity genes and generate patient-specific obesity models Fundamental Knowledge • Gene identification and function • Genetic modifiers • Molecular targets and networks Translational Benefits * New diagnostic information for patients * Insights into disease pathology * Highlight targets for drug discovery Phenotypic screen for obesity related genes - Buoyancy-based fat assay wild-type larvae No. of floating larvae 0 0 0 5 9 10 10 ) 5 7.5 8.8 9 4 10 15 20 u J— w_ V— Knock-down of the Drosophila glucagon ortholog (akh) or its receptor (ofchR) by RNA-interference(Rf) increases fat levels while knockdown of Drosophila perilipin ortholog (lsd-2) decreases fat levels Knock-down of Drosophila orthologs of candidate genes causing obesity in human patients recapitulates the obesity phenotype as assessed by the buoyancy-based assay i 1 Iii tubutin-GAL4> J> $T (Ubiquitous driver) & ^ ^ IB .1. I .tcř «> Drosophila orthologs of genes which have not been mplicatedin human obesity before Vpsl3B Rab23 C-2 C-3 C-A Vacuolar protein sorting 138. post Golgi membrane traflieing Rab protein signal transduction single-minded, central nervous system development G protein-coupled receptor brain homeobox protein, neuron fate commitment DNA binding, Orcadian rhythm DNA replication and repair endonuclease leucine-rich repeat-containing G protein-coupled receptor S-hydroxytryptamine (serotonin) receptor NAD dependent histone deacetylase activity Many novel human variants from the Genetics of Obesity Study map to identical/conserved amino acid residues in _ Drosophila ■ -- ■»■-i_J„ M7M KU2J i c* i. i Further validation (TAG) estimation. ■ Assess obesity phenotype Future Perspectives of adiposity phenotype with other methods such as Triacylglycerol incorporate patient-specific genetic mutations Elucidate the role of the selected candidate genes in energy homeostasis. Identify expression and functional domains for the candidate genes _ Rescue experiments with ; human or Drosophila gene adult flies by ubiquitous knock-down of candidate genes. using CRI5PR/Cas9 and assess adiposity, the physiological regulation of Parallel experiments in mammalian cell lines Identifying molecular targets of candidate genes Puniversity of cambkiix.i CANCER RESEARCH UK Gurdon INSTITUTE ,^fc_X_ o äi'U' fnserm ti J. 'USIAS « O, lj 5jtt* Universite de Strasbourg Impacts of Innate Immunity against Tumors in Drosophila Roychowdhury Arghyashree^, Prakash Pragya1, Goto AkiraH Hoffmann Jules" i Universite de Strasbourg, CNRS, RIDI UPR 9022, 67000 Strasbourg, France 2INSERM (Institut National de la Santa et de la Recherche Médicale) 3 University of Strasbourg Institute for Advanced Studies, Universite de Strasbourg, France Abstract Flies have been well known in recognizing and responding to microbial invasions. These reactions include innate immune responses against various pathogens such as bacteria, fungi and viruses. Apart from this well known classical phenomena, there also have been recent reports of //f V(>(| emkbU9 antitumor defense in Drosophilo larvae . Despite considerable advances cancer treatment remains suboptimal, underlining the need for new models with which to tackle the multifaceted disease. The data although still fragmentary, a point to potential involvement of signaling pathways that mediate the interactions between tumor cells and innate immune system namely JNK pathway, the JAK-STAT pathway, the TNF pathway, the Toll and IMD pathways. My aim is to elucidate the innate immune responses induced against oncogenic cells using adult fly as a model. A Drosophila Ras[V12}-GFP oncogenic cell line was used. I established an in vivo system by transplanting these oncogenic cells into the adult fly. Further, to have a better understanding of the response, I used flies deficient for the Toll and IMD pathway and examined its proliferation. Disappearance of the oncogenic Ras[V12)-GFP cells was observed in wild type and IMD mutants, but not in Toll mutant flies. The data pointed out that the Toll ' Oout^otm-izsh pathway seems to be involved in clearing off the transplanted cells in Drosophilo. , Noyo»ei rMton To get further molecular insights, I examined the role of the Toll pathway in an in vitro cell culture system. Consistent with the in vivo data, the Toll pathway activation significantly hinders the proliferation of oncogenic Ras(V12}-GFP cells but not of Drosophila S2 cells in culture. Cell (Simcox et al2008) competition showed suppressed proliferation of Ras(V12}-GFP after co-culturing Toll over-expressed S2 cells.. The effect is cell non-autonomous. Taken together, these results suggest a potential anti-proliferative role of the Toll pathway to oncogenic Ras{V12)-GFP in adult flies. Ras[V12] cell : Background ■"■VAcanG* UAS- J In vivo { The Toll pathway is required for clearance of oncogenic Ras[V12] cells in adult flies \ A ProlifCTationofGFPmarkcdRasv,:!cclls B GFP expression level ■ ♦ ? , a. ////// Toll Expression level % of the Rasvi- GFP cell proliferation in Toll OE flies 1 f,0íj i—" ^ -i' Wi lüü W Transplantation of Ras V121 cells in ro ^uti^-u .. > appearance and,,-appease 0thelLnlu*"" M>'D8S™1**' <™ P=»»a, mutant) o« to „uctily the lev,, „, J? "' „'and Z7,cT .T"" °' T°" mu,M' +** '"d | Ihs maintained at 22 c ' "'"Plantation. |C| Survival test of transplanted the RM. Tumor developed „„s afong ZI ZZZl™ „1™'°'*™°' " "C T°" • vitro G "»"''-GrPceHprolircm H S2-GFPccll proliferation Toll expression level Da> o SI-CFP cdli tnMisfecierf »iih control ■»] Toll OE I'.- 2-4 cell fflr*cr monitored Dre expression level mixed m defin.i* rtixx L Proliferation of Rasvl--GFPCclls i ""tnom^j» h" ««t on ih. '"""«"at'on count ol the sir.™ „ '""h0" The cells „er, <^«»al»n^„„«uu™ >^^a,„„ „, th, « «»• To» 0€. u„,lte fh ^—-,--»10^«,^^^«^« I Lev., o. To,, and ,------"MV12l«ll«n" «W to WS Ca- ,__™* P ° * hours PO« transfect^ („ ^ ^ „ Photosensitized Methylene Blue Inhibits Self-Assembly of P-Amyloid Peptides in vitro and Drosophila Model Systems Yoon Seok Sun12, Manivannan Subramanian12, Byung II Lee3, You Jung Chung3, Chan Beum Park3, and Kweon Yu1>2 1Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea 2Korea Institute of Science and Technology, Seoul, Republic of Korea 3Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea Abstract Abnormal aggregation of p-amyloid (Af» peptides is a major hallmark of Alzheimer's disease (AD). In spite of numerous attempts to prevent the (5-amyloidosis. no effective drugs for treating AD have been .eveloped to date. Among many candidate chemicals, methylene blue (MB) has proved its therapeutic potential for AD in a number of in vitro and in vivo studies; but the result of recent clinical trials performed with MB and its derivative was negative. Here, with the aid of multiple photochemical analyses, we first report that photo-excited MB molecules can block A|3 aggregation in vitro. Furthermore, our in vivo study using Drosophila AD model demonstrates that photo-excited MB is highly effective in suppressing synaptic toxicity, resulting in a reduced damage to the neuromuscular junction (NMJ). an enhanced locomotion, and decreased vacuole in the brain. Our work suggests that light illumination can provide an opportunity to boost the efficacies of MB toward photodynamic therapy of AD in future. Summary Schematic representation showing inhibition of AP aggregation by photo-excited MB. The in vitro and in vivo experiments performed with the Drosophila AD model were conducted under the illumination of red LED light. The binding interaction of MB to Af) aggregates and the pnoo-oxidation of the peptides induce disruption in the structural conformation, thereby blocking (or reversing) the progress of aggregation. Dark Light 2M 210 Z» 2» Wavelength / nm Dark Light Aß only w/MB Dark Light 9 Figu : aggr. None treated aht-lnduc.d tWUIPT"— <* ¥ •»» MB' <"> CD,05^asU,e £ S ^itfTllm vanoub condition., (b) ThT fluorescence assay to measure me ,c Srtve.-stai."*! naive gal electrophoresis showing Inal the amyloid tionls icj oi mb "J- undw lign, nomination The arrow onvonts »as J*"*"* |;0,fesporMlli to ^ monomer, ol AO (d-g) Rep.esenlat.ve I All" "V fa with or wilhoul MB under darti and light conditions Figure 2. Photo-excited MB reduces the brain vacuolization in adult Drosophila. (a. Representative haematoxylin and eosin staining of adult head sections in AD model flies {elav>Aß4: with or without 100 nM MB treatment under dark and red LED light conditions. Arrows indicate vacuole phenotypes in aged fly head. Scale bar: 20 pm. (e) Quantification of the vacuole size in adult head sections in AD model flies (elav>Aß42) with or without MB treatment under dark and red LED light conditions. öle F(o_ 3 Pho,M,e,..d MB -stores «. pn-no^, of Aft J-*r^^^5 JSC ' cSTriTESSi on muse* ^'^ZTTJ^^^ - <* 9> MB treatment under dark and red LED jghL <**» . „, aI)d „. h) NMJ of the Uhc'AB42 under dark condition: (b. d) ^"J^,^-**** . -omnokjgy phenolvpe caused by A042 seated with or without 100 nM MB ^<^„LED^h' fl,, «^«^7^ cocentrabon of Impression ,s rescued by photo-excited MB. jScate bar 10 pm 0^ ^ ^ MB on mental number of NMJ boutons on muscle 017 of A3 ^>™J^m. resp**«ly (l>_n» MB on me lo Diameter of inner-circles are 1.0 cm. 2.0 ""T |H)MMe „šuměn, under ,ne^rL°h onhHEoStS* controt and (k) Mhc>A8« ««hor 90 seconds demandLSDfcht Scale bar 2 cm (!) QuanWcaboo of crawted d^Bnc of , pnoto-exced MB motecules exhibit a high degree of inM*on ag-"* fr-ny*«*» » l*» _„^^»u>en R35V12 Whole RasV12 Proximal ■ l_ m overexpression in the SGs activated two stress response pathways, JNK and /STAT. JNK (Left panel: green) is predominantly induced in the distal part and JAK/STAT ;ht panel: green) in the proximal, assessed by the expression of their respective orter constructs. In the distal part, hemocytes (red) are recruited to the region where JNK pathway is also induced. UPD2 Socs36E To verify the readout from the JNK and JAK/STAT reporter constructs, whole, distal and proximal 120 h AEO SG were dissected. qPCR was performed for known JNK (hid, TIMP, MMP1 and UPD2) and JAK/STAT (Socs36E) target genes. In line with reporter constructs, JNK target genes (TIMP, MMP1, and UPD2) were expressed higher in the distal part of the SG in comparison to the proximal regron. Moreover, JAK/STAT target genes (Socs36E) had a higher expression pattern in the proximal region whereas in the distal, the expression was lower. These results confirm that there is a differential regulation of the two stress response pathways. RasV12 mediates action of Drone (caspase), a down-stream component of JNK pathway osophiia JNK-pathway is known to mediate apoptosis, ROS production, and recruitment of hemocytes. Therefore, we asked whether JNK caused induction of apoptosis in the distal part of me RasV12 salivary gland and whether JNK is the apoptotic inducer in our model ruas (TRE: green) expression pattern as shown previoH^sexp^SH * »* mZ^J^^V! ^ RaW12 * Kn0cWn«-do*n Bsk "duces expression Thus uJT HOWeVCr' Oronc ac,ivit* "as 3|S° (tetKttd in the proximal ^^Tl "r^™ "eeded'° ***** WWfc distnbuuon NeveZL ^^TZ^ZT°t0il " *mo"""-' "V «he genetic °< ROS" * l cl.r* on the red Su* ^ button. Suflgest a Topic for a Video Please contact FlyBase if you have Mfi-The Contact FlyBase' form can &^Bm every FlyBase page. Existing Video Tutorials Basic Navigation & Tools • How to find all data related to 3 gene • How to generate an excel file of all alleles of a gene • Finding related genes In FlyBase: Gene Groups • Finding related genes/alleleslff FlyBase; Vocabularies • Finding related genes In FlyBase: The Gene Ontology • Using the Orthology search tool RNA Seq Series • Part I: Using GBrowse • Part III: Searching for Similarly Expressed Genes for a new video, at the bottom of flyBase Guidelines • How to Cite FlyBase • Authors Guidelines •TWrlyiaMConMrUun, |QC Hoben Pt-mmon, Cattmn Gene Snapshots Gene Snapshots are short summaries solicited from an expert researcher, curated and standardized by FlyBase, that provide a quick overview of the function of a gene product. For now these are limited to protein-coding genes and appear at the top of each D. mel-anogaster gene report. Gene Snapshots are also downloadable via the Batch Download tool and our precomputed files page to aid in genome-wide analyses and screens. 3' *JH. WMiH fatoary 14.1017 Gene Dmer\Egfr ; FlyBase Home Toots Downloads Links Community Species About Help Archiv Symbol Egfr Í55-^ D iiiiebuiüoeifer Name Epidermal growth factor receptor AnnoMbon .yrnboj CG10079 Futura typ« protMn ooolng_gana FryBaaa'D F8gn0OO3731 Gar» Vod.. Statu a euren) Stock availability S3 pübWy avaáabW AJ.o Known At OER. Up. Ab. Elp uEGFH, Egf-f c-a-BB Cana S napa hal EpWamad growth '»clor racaptor |Egfr} J ir» TGFa family (Brk. apt, vn. and Km). whk» ui ragutat-on. call turvrval and developmental p tr on »membra - o tyroaina kinase nacaptor for Mgnaeng IganS* in me aaaa lha inlr>caii.iar MAP Una** pdlTwey Ejtr rotas nduM grmrth ■öam>ng [Das* leal renewed 2016-10-06; Help FlyBase Produce More Gene Summaries •Thanks to the great response from the community, as of the FB2017_01 FlyBase release, there are 1901 gene summaries available in the database. • However, many genes are still lacking their Snapshot and we very much welcome user contributions! • If there currently is no Snapshot available for your subject gene, please send us your suggested summary via a standardized form accessible either directly from the gene page (via the 'Contributions welcome' hyperlink in the Gene Snapshot field) or from the Gene Snapshot wlkl page via the Community button menu at the top banner of every FlyBase page. FlyBase A Dal*Da»e of Drosopham Genet & Genome. Mirygold, Gillian Millbum. Glulla Anlonanu, Helen Attrili, Sim* ■'HI, Lynn Crosby, Gil dot Se-.it«», Oavld Emmait, L. Stan Grama Goodman, Gary Grumbling, Vitlur 5lral*t%, Jim Thurmond 0 "•<-• KeMtarcri Institute at the U.S. National Inetllutea of Heel'' Initiative, and the National Science Foundation Wire ov§, Imam Garapati, Ale* Holmes, Tannin Jonet, Aoife Larkin, Ailf ojw. WQf TrovlSCO, Jose M. Urtano (FlyBese-CambnOge), mrhteen Falls, Beverley Matthews, Susan Rvtso, Christopher J^aL Pinglei Zhou. Mark ZytkovK* (FfyBeteHervant). >>■■<: Indiana), Mchanj Cnpps, Maggie Werner-Washburne, Phillip lke't (FlyBase NetsHexk .< ■ IH1HG000739 )- Support n also provided by the Brltlah. Medical mir-1- Council (G1000968), the Indiana Genomic* 1 *"iEDE resource* provided by Indiana University. FlyBase CTTCATAGATl fcCATATATATW Finding the right tool for the job: new ways to find particular types of transgenic construct in FlyBase Gillian Millburn (gmll9@cam.ac.uk) and the FlyBase Consortium The long history of fty research plus the sophisticated range of applicable genetic engineering techniques mean that a large number of increasingly complex transgenic fly lines have been generated and described in the literature. While this rich genetic tool-kit helps to make Drosophila melanogoster an ideal model organism to answer a wide range of biological questions, it also creates a potential problem • how to find the most appropriate fry line for a particular experiment from the targe set that is available. To help overcome this issue, FIvBase will start to capture and display information about Experimental Tools, which will allow you to easily identify transgenic constructs and their insertions with particular characteristics. We are defining an experimental tool as "any sequence whose own biologicol function isn't really being studied in an experiment, but is instead being exploited to study the biological function of some other gene product or a biological process". This broad definition will allow you to browse and search for tools used for a wide range of different purposes, such as enabling a gene product to be detected (e.g. FLAG tag, GFP, other reporters), targeting a gene product somewhere specific within a cell (e.g. nudear localisation signal, signal sequence), driving expression (e.g. GAL4, lexA), enabling clonal/conditional expression (e.g. FLP, FRT), being used as a sensor for changes in CV\ pH, voltage Experimental Tool Report 'Experimental Tool Uses' Term Report Uses term(s) describe what the tool is used for. * Crick on the term to go the Uses Term Report Related experimental tools section make it easy to jump to reports for related tools at a finer level of detail than the "Uses* section. • Click on the Tool name to go to its Tool Report • Click on the Uses to go to the corresponding Term Report Columns list the components that make up the transgenic construct Lask to relevant Gene Vv^e tr* main gene product ts a tod. column* list Tool \lGfP) and us Uses {e.g. ftiM) or Tool (e.g. UAS] report \ Additional Toonst [e.g. FRT) in the construO that do not form pan of the encoded product y- Mairi branches of Uses tree and examples of Tools ■» descriptor L binary expression system \. gene product activity regulation ag 5 gene product cleavage tag 9 gene product degradation tag T: miGFPi. T:PE5T-MmuiVOdc : gene product detection tag RNA detecOon tag B protein detection tag epttopetag T:MYC. T:FLAG. T:po*yHis 3 binary expression system um weit emary expression system - armer GAU. GAU::VP 16, Of bnary expression system - regulatory region UAS. QUAS binary express.cn system - repmacr GA180. QS Click on any column _ ■ -~H heading to reorder the Ubtes Where the function of the encoded product a bong studied, columns hst the encoded Gene (rf>eo| and its Product Class (*tffl-tffie gene product) List of Toots U6TP) that are fused u the main gene product, plus their Uses (green flvoirseeni prolan) reporter enzyme lacZ. pLUC r gene product localization tag iMyt StxAtB. TaJa-tra 3 genetically encoded sensor mechanical force sensor pH sensor pHluoeinE redox state sensor purification ag TpoIyHis site-speonc recombtnas* FLP, FLPmS.cre Bte-speeme recomtwiaOon target region FRT. mFRT71. loiP r spat system component Ml EBFP cyan fluorescent proceei Cm*an. CFP far-red fluorescent protein mKate green fluorescent protem EGFP. GFP. PA GFP 3 moouia-aoie fluorescent protein v protein SiOwFT photoactwatable fluorescent proton PA-GFP ■rotem Oronpj rasa mKO mOktrry, tdTorruto yeaca* fluorescent protein EVFP. Venus r bzz- P . _ 1 _ — . .. f— t -1- ■ a—fi . . insertion into tHertappmg genes Transgenic Product Class' Term Report ^ he Transgenic Product Class is intended to give an overview of the nature of the gene product encoded by a transgenic construct, similar to Allele Class used for classfcul and insernonal alleles. * ft Mfifi be used for constructs where the function of the encoded gene product is being studied or where the construct encodes a sequence that affects an endogenous gene product (e.g. RNAi) • It will allow constructs to be grouped into broad categories for browsing and searching, for example in the Transgenic Constructs laoles on trie Too) Report and in similar tables on the Gene Report (see below]. New terms can be added to describe other kinds of tool as new techniques and toots are developed. Current ideas to expand the list of terms include: • 'genome engineering' tools e.g. Cas9 * neuron activation/inhibition tools e.g. channel rhodopiins soil system component spat Omer - PNA-tm*no fragment GAL40fJO-Oo-scM ceiver - transcription aCOvaoon fragment VP1&AD-spfct fluorescent proceň C - Venus. V Venus latZ-fla. tacZ-Aüt We have recently introduced a GAL4 etc' search, but it is currently limited to searching for a small number of common drivers {GAL4, QF, lexA) and reporters (tocZ GfP). Click on »rry column hradmg lo reorder the table Once we have introduced the Tool Reports and Tool Uses terms we will be able to expand this search as illustrated below. 1. The search can be expanded to include any type of toot Typ-ig oi the Tool Uses ben ma bring up afl the matching Uses terms, e.g. typing J*uor* ^ • • MOCK UP * • >g put* v ft are a work-in progress wild type grne product knockdown of gene product AMA> 2. Vou will be able to use the regulatory region informanon to search for tools whoi expression pattern reflects that of a Orxe you rwvr selected t pjrtxuUi Use, I a «opdown ai the Tool ben on the nght wWI show ** the corieipono^nj loan >aMU< IhrBufS* XSIQ4 'cvw>n pMwa ky waunu Uruvxuty j» UNIVERSITY OF T CAMHR1DGE orma ictomyosin cables 'ifnif ini-i'i a«nui.^JTMjj!t,*ii •ts> IT *--—■ ■.. . ■ • Confirm w • Assess pre • Assess mo iether the cable of th ■ence of sarcomere-as ecular organisation of salivary gland placode is absent in the ZjsPS2A mutant sociated proteins in the cable actomyosin in the cable Generation of non-centrosomal microtubules promotes apical-medial \ ^kWWW actomyosin accumulation during tube morphogenesis in Drosophila MRC' "°'^B°00'' Ghislain Gillard, Gemma C. Girdler, Alexander J.R. Booth, & Katja Roper MRC Laboratory of Molecular Biology. Cambridge Biomedical Campus, Francis Crick Ave, Cambridge CB2 OQH, UK. Salivary gland (SG) morphogenesis INTRODUCTION Most of our organs, including all the respiratory, secretory and circulatory organs, are composed of tubes. Using Drosophila embryos, we are investigating tube formation during salivary gland morphogenesis. SG formation begins at embryonic stage 11 with invagination of two sets of ectodermal cells, this invagination being due to apical constriction that change the cell morphology into a wedge-like shape. Proper apical constriction relies on the accumulation of actomyosin at the apical-medial domain. Surprisingly, whereas the role of the actomyosin cytoskeleton in morphogenesis has been extensively studied, little is known about the role of MTs in morphogenesis. However, recent work in the lab has highlighted a role for non-centrosomal MTs (ncMTs) in apical-constriction by promoting apical-medial recruitment of actomyosin [1]. I will present here a project and preliminary results aiming to identify first how these ncMTs are formed then how they promote apical constriction. Apical-medial recruitment of actomyosin Formation of ncMTs during SG morphogenesis _ Early stage 11 Mid stage 11 late stage 11 ncMTS are required for medial actomyosin accumulation 1 ■1 V BE* Ks c In. ■ ~. 1 ■< \ . > UtrophinGFP Hfl B' I H w mm Membrane U'ni-mGFp Membrane ■ '■ • Assess the effect of Patronin and Ninein depletion: Who comes first? Live imaging of individual components of the hub '40: lagenivin cell intercalation Audrey Placenti, Delphine Cerezo, Marilyne Malbouyres, Florence Ruggiero, Stephane Noselli and Veronioue | UNIVERSITfci CÖTE D'AZUR :T /an De Bor w Introduction Basement membranes (BMs) form an essential extracellular protein meshwork, holding cells together. BMs are quite diverse and their composition defines organ biomechanical properties,. * They have essential role in different cellular be|^p migration, polarity, cell survival and differentiation."-' molecular mechanisms underlying these fur signalling pathways involved are poorly ch these questions we study how BM control! process leading to the inter-follicular Drosophi/a's ovaries. V£F ■ 1 L_ i The Role of Sidekick at Apical Vertices in Epithelial Morphogenesh Tara Finegan1, Nathan Hervieux1, Alexander Fletcher2, Guy Blanchard1 and Benedicte Sanson1. Department of Physiology. Development and Neuroscience, University of Cambridge, UK. ^IVERSITY OF * -'School of Mathematics & Statistics & The Bateson Centre. University of Sheffield, UK. 3*Z PA IVIRR IHPF E mail' tmf32@cam ac uk nh480@camac.uk. a.g.fletcher@sheftield.ac.uk, gb288@cam.ac.uk. bs251 ©cam.ac.uk v 1 We use the early Drosophila embryo as a model for tissue elongation Orosophita embryos extend their body axis in a process called germ band extension (GBE) using a common morphogenetic process found in animal development: convergence and extension. EXTRINSIC PULL In vagina I log tltiue pulls germ band Ventral cell intercalation l eal lln other epitheha adherens junctions: No vertex specific proteins found in the early-embryo during GBE (no mature septate junctions). scpl.il c junctions: ■ GliOl.lClln (Schulte et al 2003) Anakonda / Bark Beeile (Byn etat 2015 S HikJetxanol et al 2016) ■ Mud {BosveSd oi al 2017) 2. Cell intercalation is driven by actomyosin activity and changes in adhesion Epithelia are polarised Intercalation occurs specifically at actomyosin enriched boundaries established by anteroposterior (AP) patterning (Tetley el al. 2016 eLile ; Pare el al. 2014 Nature). i Parasegi AclOmyosin enrichments Proteins localised at apical adherens junctions drive intercalation. Lecuit. Lenne. Wieschaus andZallen labs Apical view ot cells intercalating in a "T1 transition": /"v. cadlwrin removed m Actin-based protrusions basally I also contribute to intercalation (Sunelal. (2017) NCB) 4 Apical view The Ig super-family protein Sidekick defines apical vertices Screening YFP-tagged proteins, we discovered that Sidekick localises specifically to vertices at the level of adherens junctions (Lye et al. 2014 Dev). Sidekick is the first protein found to localise at vertices at the level of adherens junctions in any epithelia. Sidekick 222.JH.1 YFP - - Intracellular M iSFV - Extracellular - Super resolution SIM imaging reveals Sdk-YFP forms strings at vertices: 5. Sdk-YFP invades shrinking junctions extracellular vertex spac* ^ Live imaging (30s / frame) Growing junctions: Supět resolut Sdk-YFP Actin Sdk-YFP 6. sdk mutants show severe intercalation defects, compensated for by an increase in cell shape changes W patterning and actomyosin enrichments are maintained in sdk. sdk ■ ** mutants show abnormal Ussue •I 4** geometry: k mutants show a delay m gbf „ «. compensated lor by an *,d a severe intercalation Jumulai-veTissue _ 0611 shape changes Strain Hale m c"'""'aftve tea/ r,lm , «„ _2 Strain flate + Cumula"^ Shape Ä * K ■ Could Sdk be playing an active role in intercalation? 7. A computational vertex model recapitulates the sdk phenotype Intrinsic processes modelled. Line tension high between cells oi different identities and active cell intercalation implemented (shorter interlaces shrink more quickly) sdk As wt but loss ol active cell intercalation and uniform extrinsic pull added 8. Sdk is required for epithelial tissue integrity during morphogenesis Epithelial holes are more common and persist for longer in sdk mutants. sdk Myosin RLC-mChcrry DE-Cadhenn::GFP Myonn RLC-mCharry DE-Cadhofin::GFP — p "00001 oi "p-0018 I-1 í I, 9. Conclusions and model 1 We discovered the lirst protein to localise to epithelial vertices at adherens junctions Sidekick 2 Sidekick is required tor correct active cell intercalation driven at the apical domain 3. sdk mulanls complete GBE because the cell intercalation detect is compensated by cell shape changes, driven by the posterior extrinsic pull. 4 Epithelial holes are formed in sdk mutants undergoing epithelial morphogenesis. Slik- adhesion and cortex discontinuities call» ,, ,, over elongated cells in a stochasti Maximilien Courgeon and Claude Desplan DepartmertcrfBi^^ r4ew York NY 10003, USA leciScayfaon off \ in the fly retina.: IHtar Tate- WRA Circadian Control of Mechanosensation Jason Somers1, Jake Cable1 Ross EF Harper12, Ariana Hiibner12, Jorg T Albert12 il " rsitv College London, London. United Kingdom Ear Institute.----^ L|fg Scjences and Experimental Biology (COMPLEX), University College London, London, United Kingdom -Cent™ for Mathematics rny [) somers@ucl.ac.uk)[ joerg.albert@ucl ac uk ] £. ■ ■ ^% I Abstract: The circadian rhythms of an animal's physiology are driven by molecular oscillations within a variety of tissues; some tissues are under hierarchical control of central pacemakers, but several tissues in Drosophila display autonomous, molecular and functional, oscillations'1. This project will investigate a similar phenomenon in chordotonal organs focussing on the fly ear, i.e. the Johnston's organ (JO), which is located in the second antennal segment (a2). The JO is a cluster of mechanosensitive neurons that transduce antennal motions into nerve impulses. Here, the daily profiles of their energy expenditure should ideally be matched to the daily profiles of their corresponding mechanosensory behaviours. Gene expression in JO Is there a molecular clock in the JO? Clock genes are expressed in the JO in phase with clock gene expression in the head Mechanosensory gene expression show no clear oscillations C) Head - Clock genes B) Anatomy of fty antenna and chordotonal neurons (C - E) qPCR results of dot* gene and mechanosensory TRP ] darnel genes expression in adult head and JO over a day. * v*ues are normalised by Z-score */. SEM for three biological Sensitivity to mechanical vibrations Mechanosensation also allows the fly to response to less subtle stimuli while also providing proprioceptive feedback during flight and locomotion. Oscillations in response to mechanical stimuli were investigated. A) Cantons - LD B) CantonS - FR D( per01 - FR E| Cantons - LD F) CamooS - FR iky, i ÉÉilÉá IE* »"-FR Pre- and post-stimuli speed comparisons reveal clock-dependent, varying levels of response throughout the day "*""» neat ol tu » CO am FR OroaapMaMtouW T IC - M) FW Clock controlled biophysics of the antennal ear Drosophila antennal ears are finely tuned to their conspecific courtship song via an active process that requires an injection of energy. As auditory dependent behaviours (e.g. courtship) exhibit circadian rhythms rhythms of antennal biophysics of the antenna were explored. Changes were observed in some parameters but not consistent in FR conditions. A) C) CantonS - LDfTC D) CantonS - FR il. ä li i ik.. i B> .Best frequency E) per" - LDÍTC F) per" - FR Zeitgeber Time G) CantonS I) CantonS Ji 3J1 Mé- H)pe/" J) per" 1ÉI » "I "ů1 ,. T s FR caodLfcn (G - H) Box-plots of extracted jim ■■■In i measured from6 separate ivnepores see (B) far wi of parameters (I - J) Median AC of eaon Wneport for both genotypes. F* uses a oarriperied harmortc o Future work Investigate mechanosensory protein levels/ localisation Biophysics assays with females White noise and/or step stimulus experiments to dissect motor/transducer contnbutton to antenna tuning Funding ^^^^ References ere More than black or vv Variable roles of the reaction in Drosophil J.P. Dudzic, D. Main, B. Lemaitre Ecole Polytechnique Federale de Lausanne (EPFL), Gl hite: ■ melanization -or immunity >bal Health Institute, Lausanne, Switzerland (Pfl jan.dudzicripepfl.ch Abstract The arthropode immune mechanism of melanization is catalyzed by enzymes called Prophenoloxidases (PPOs)1. After an initial stimulus these PPOs are activated by a cascade of serine proteases (SPs)2. We investigated the role of two SPs previously connected to melanization" Hayan and Sp73A. We found two distinct modes of melanization depending on the involved SP Hayan is involved in the local melanization of the wound site, while Sp7 dependent melanization is necessary for the clearance of septic infections. 1 F'g-1 win 8 Hayan* J*> BN SSK H* Fig.1: Clean injury confirms alternating phenotypes in two serine proteases involved in melanization. Wild-type animals show rapid melanization of wound sites. Hayan mutant flies show a loss of melanization (arrow) while larval melanization is only partially impaired. Flies mutant for Sp7 show an alternating pnenotype: melanization in adult flies is not impaired while larvae show a complete loss of melanin deposition (arrow). The PP01,2,3 mutant acts as control for the total loss of melanization. Fig.2: Survival experiments with S. aureus show susceptibility of Sp7 mutants. While wound melanization seems intact in adults, Sp7 mutants phenocopy the survival defect of melanization deficient flies. In contrast: Hayan flies which do not show wound melanization show no survival defect. Flies with defective IMD or Toll pathway are not susceptibility against S. aureus revealing a melanization specific pnenotype. 5. aureus OD 0.5 Fig.3: Infection with fluorescent S. aureus reveals that Sp7 mutants can not control infections albeit wound melanization seems not affected. Hayan mutants clear infections quickly although wound melanization is not apparent. Flies mutant for melanization {PP01,2,3) show the same Fig. 4 ■nization reaction can be assumed by the shown evi-r melanization of wound sites in adult flies, i.does Conclusion: Two distinct types of the mela dence. While Hayan seems essential for proper rneiejni«auuii wvvnw *■>«<» ••■^~~-tho rnnfrarv not show any defect in the melanization dependent clearance of S. aureus Sp7on tneconndy seems to be dispensable for the melanization of wound areas, but is mandatory, for surviving S.aureus infections in adult flies. Elements of the Toll pathway fCtt^tlr^ase^lMOT » involved in the differential activation of those two, non-redundant melan.zation functions, wnicn are dependent on the activating SP. ■^^^^■^■j Fig.4: Gram positive M. luteus can induce wound melanization in absence of Hayan compared to clean injury wounding (see Fig. 1), when suggests a role of the Toff pathway irj melanizatron. Combined melanization and survival experiments reveal further evidence for the Toll narhwav activating serine protease ModSP which ffirorlnch tog the melanization pathway to TcfcateSp7 dependent clearance of S. aureus. Functional characterization of Drosophila adenosine deaminase-like protein (ADAL) in embryogenesis Yu-Hsien Lin1, Michal Zurovec2 Faculty of Science, South Bohemia University, Czechia Entomology Institute, Biology Centre CAS, Czechia (r99632012@gmail.com1; zurovec@entu.cas.cz2) Introduction adenosine deaminase-like protein (ADAL) has been shown to be involved in the metabolism of N6 purine nucleoside monophosphates in humans (Murakami et al., 2011; Schinkmanova et al., 2006). However, t' physiological roles of ADAL are still unclear in any kind of model organism. In the present study, we generated a nu adal mutant by CRISPR/Cas9 approach and described the function of Drosophila adal gene (CG11194) in Drosophil embryogenesis. or 06- substitute adal mutagenesis by CRISPR gDNA deletion Absence of gene transcription N.s N.S adal [Pbac] RNA-seq values (Flybase FB2015JJ1) ada) [pbac] Adapted from BDGP database 20 Conclusion Ladal loss of function significantly reduced embryo viability (-16% hatch rate) 2. The reduced hatch rate of adal mutant eggs fertilized by wild-type males indicates that adal is maternally provided 3. adal mutant embryogenesis fails to proceed the early nuclear division. Reference t Murakami E at al 20it Adenosine deamina»e-luie protein j ^ . ______fc,,,,, ,n tfajt twriroivaa Ot tMSh Of Oft and substrata speoncity in me nyoruiy»w u* "'"r ' aminopunne nucleoside monophosphates J Med 2 Schinkmanova M et al 2006 No-methyi-AMP a^^™T*, purine acyclic nucleotide phosphonates Biochem Pnamiaoot itas Nti-*uD*titurea Revealing the interplay between Hedgehog sipnaling and metabolism during growth control of the Drosophila melanogaster wing disc lognnis Nellas', Venkatesan Iyer', Stephanie Spannl', Stefanie Schirmeier2, Suzanne Eaton' 1 Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstralse 108, 01307, Dresden, Germany 1 University of Miinster, Institute of Neuro- and Behavioral Biology, Badestrafie 9,48149, Miinster, Germany Introduction I Hedgehog (Hh) proteins regulate growth and patterning during development and regeneration. Tissue differentiation is controlled by the establishment of g. expression patterns mediated by the Gli family transcription factors. While the pattern of differentiation centrolled by morphogens has been well charactpp, we lack the understanding of how they promote growth during normal development. Metabolic shifts are key drivers of growth during tumorigenesi-, I whether metabolic regulation plays a role during normal growth is not clear. Here we investigate the role of Hh signaling in controlling specific pattern, metabolic activity during growth in the wing imaginal disc of Drosophila melanogaster. Cells at the organizer regions have unique metabolic properties and need to define and understand how metabolic patterning contributes to the growth of the wing disc. Scientific questions Results How does morphogen signaling interface metabolism? What is the metabolic state induced by Hh signal in the A/P boundary? lso< nr.in- dehydrogenase G6PD Aldehyde oxidase T W !■ Kuhn and Cunningham 1986,1987 A/P boundary-like pattern of metabolic enzymes (!) Does Hh or other morphogens regulate their expression or activity? What is the role of metabolic patterning in growth control?_ 2 ) Knock down of 6APDH2 activates Hedgehog signaling in the wing disc. Ap-Gal4>Gapdh2RNA1 1 \Gapdh2 KD elevates the levels of the signal transducer Smoothened and Ci. ^Mitochondrial membrane potential is stable throughout the wing disc pouch in wild type flies, as indicated by the TMRE staining. Study goal My aim is to study the interplay between Hh signaling and metabolism in growth control of the Drosophila wing disc and reveal the metabolic status of cells at the A/P boundary where Hh signal takes place. Experimental approach Hh Metabolism Metabolic enzyme levels and localization • Monitor protein levels of GFP-protein fusions • Localization after Hh perturbation Genetically encoded FRET sensors from reporter flies ATP, lactate, pyruvate and glucose Effect of Hh perturbation on metabolite levels? Metabolites level Metabolite m-iimiij/ ilum.iiii A(lj|.in(l liuin Sun Martin ri at, 2011 Metabolism I tudbdck on murphugun signaling * UAi/GdW sy.tem to knockdown metabolic enzymes uijIhk KNAI • Immunohlstuthemislry tur Hh signal components (PK, Smo, (!,,,,) g ) ATP levels are mostly homogeneous throughout the wing disc pouch. Color code: High ATP levels H | Low ATP levels y\ Distribution of metabolites {glucose and lactate) from glycolysis appears to ,_J be mostly homogeneous throughout the rmaginal wing disc pouch. > MMP and ATP levels are homogeneous throughout the wing disc pouch, so do some metabolites. Thus, which aspect of metabolism changes? What is the contribution of the metabolic patterning in growth control of the wing disc? How are these regions metabolically different? Once we understand how they in different, we can ask what happens to the remaining tissue compartments of the wing disci References Altjandro tan Martin ft at. plos one. 2014, 9(1): es$780. Altjandra ||n Martin et at. PtoS One. 2013;8(2):ii-57712. Sprt-y .nid Kuhn. Genullt.. 19«/ Feb, U5(2):283-94, Kuhn and Cunningham. P«v. Gwnet. .986,7<1):2134. jTDIGS-BB CBG Mil* fl*.t. » IntHtwM Jl 133 UPPSALA IN1VERSJTET Gelatin and Starch Media Stabilize Bacterial Luciferase and Oxidoreductase Anna BEZRUKIKH1. Elena ESIMBEKOVA21, Valentina KRATASYUK1-2 Mb 1 Siberian Federal University, pr. Svobodnyi 79, Krasnoyarsk, 660041, Russian Federation, Aebezrukih@gmail.com 2 Institute of Biophysics SB RAS, Akademgorodok, Krasnoyarsk, 660036, Russian Federation The aim of the work was: To examine the effect of the viscous getatin and starch i rrwroenvironments on the stabSty of coupled enzyme system of tunanous bacteria NADH:FMN-o»doredLKa^ (R*L) whan exposed to various physical and chemical environmental factors. Results: 1. Ading of RH. to getatin soin-atesbio^^ whie including of RH in the gelatin gel rriatrix increases its activity. For instance, at 20°C in case of 1% gelatin and at 25°C in case of 5% gelatin the luminescence intensity increases by two times. In the presence of starch the biotunmescence intensity of RH barely ^■an^es ejMl '.eT^r^"'...^ var*.at>c~ = '.= Tc"2:.."5 remains 25°C. 2. Thermal inactivabon of RH. includes two stages, which correspond to the processes of enzyme dissociation into subunis and ther subsequent denatuattoa 3. Getatin and starch viscous riwjoefTVTonrTients do not change the thermal lTacSvabon rate ooratatte of RH. in a temperature range torn 23 to 43*C. Reactions catalyzed by oxidoreductase (R) and luciferase (L): R NADU + H* + FMN -» NAD* * FMNH- FMNHj + RCHO + Oj -» FMN + RCOOH + H-0 Methods: The thermal StabSty of RH in a getatin and starch corfcweiu media and the effect of gelatin and starch on the stabSty of RH under -IM-~l--^--1.......■ r< , :r_,r r^pnjff) „.....^tfM j A reagent composed of RH irnmobSzed in gstatin gel by dosing in Therein litJJKJivatkjti IdneBc curvet of RH at uineieiit terjii|ieieture» In the presence of 1% gelatin (b) and In Its absence (a) Dependence of the luminescence intensity of RH on temperature in buffer solution (control) and in the presence of 1% or 5% jjiliai, 2% starch t ottoefcet (*J end second gjj t •ofR>t.toe^Me*»nr«oM%uilill andtn M far a ■Win T.«C • «r»»0I. »««r' ZS KM ' 0.SWU2 25 • 4»C f ■jaj«e) i.*=as OjtttCJ » >»i »1 -«•=• • «3 »i MM »* Sit'* 4. RH. stranded by bet resistance in an aksfew pH t 5. Gatetin «,.i.jtofeec res-*': ■ two years when stored at 4*C -5 ar=v-> for iosciences Opsin-based photorecepnon in á 5X^C luminous brittle-star Jerdme Delroisse l, E. Ullrich-Liiter2,0. Ortega-Martinez3, J. Mallefet *, P. Flammang 1 M Organism Biology ft B>omimet.cs Ub, UMONS, Belg.um ; 2 Muieum fur Naturtunde, Berlin, Germany, 3 Departmem of fcolewcal and Environment*] So*"". Unrverstty of Gothenburg, VJantwtbtit, Sweden . 4 Marine Biology lab, UCL. Louvain-U-Neuve. Belgium Introduction IExtraocular photoreception in Echinoderms In metazoans, opsins are photosensitive proteins involved in both vision and non-visual photoreception m. Echinoderms are no exception to the rule even though they havenoeyesp, Sea urchin genome : 8 opsins genes of which 4 are homologous to metazoan visual opsin 13] No data for other echinoderm classes! J Case of a luminous brittle star... -Project focused on the luminous brittle .star ?. Amohiurafiliformis Iblueemission) M1. ' >f \ Infaunal ophiuroid, large densities on soft bottoms. .' . Co"espoodfng»oihor: Bioluminescence & Extraocular photoreception <* Molecular markers of photoreception were identified in the light-producing organs of 1 a sepiolid squid [6, and a ctenophore m, suggesting a link between bioluminewnrg and Photoreception in these phylogenetically distant organisms. Extraocular photoreception would constitute an adequate control of photoeenesjj J these species. I v- Could such a mechanism be present in ophiuroids? Opsin diversity estimation in the luminous brittle star A. filiformis 4- "Classical opsins" (rhabdomeric and ciliary) immunodetections -fr Functional link between bioluminescence and photorecepnon? L Organism collection ■ GuUrnar Fjord • Kristjneberg Marine Station (Sweden) 5. Whole-mount immunofluorescence Anti-[sea urchin rhabdomeric 00501}*** Anti-fsea urchin ciliary opsin]Ats ?Antnxr7yiolret cdpho-tvbtJxi Abi (Uenrnt system! C- opsin & R-op sin 'sL**** ' es in the purple sea urchin (Strongylocentrot 2 homologous Sp opsinl (encephalopsin group) Development of a highly sensitive and rapid chemiluminescent assay for hydrogen sulfide Showa University, School of Pharmacy OHidetoshi Arakawa, Chiaki Nishiji na [Abstract] Hydrogen sulfide (H.Si is aiiracling attention as one of liircc cndogemiusly generated gaseous signaling compounds, the olhcrs being carbon monoxide ami nitric oxide. Mie hydrogen sulfide in live cells is generated by the following three enzymes: cystathionine |i-synthase (CBS):cyslathioninc y-lyasc (CS!i);and .l-mcreaplopynivalc siilfurtransferase (SMST). These enzymes arc involved in ncuroirnnsmiiicr regulation and vasodilatation. However, hydrogen sulfide, the odorous component of waste and sewage, is a tonic gas; therefore, a highly sensitive and specific method for monitoring H:S is desired in order to protect human health and the environment.' Hydrogen sulfide is generally measured by gas chromatography, hul this method require* special equipment. Fluorescent probes for hydrogen sulfide have also been recently developed as a simpler method. In order 10 analyze hydrogen sulfide rapidly ami sensitively, we have developed a novel method using lucigenin chcmiluminescence in the presence of copper ion (II). Cu2+concentration ESR spectrum 0 1 W/LH i .5. MfcnmofA OO, SCO 0 lmoVUM*A C»> Luc*'7 x O2 * x Luc(OO) Standard Curve for Na2S 1082 11.29 100 1000 Phoiphile BuHer p H Assay method Lucigenin chemiluminescent solution (0.2 mL: 5 umol/l copper chloride (11). 0.04 mg/mL lucigenin. 0.1 mg/mLTritonX-100) was added lo Na2S solution (20 ul) diluted with phosphate huffcr (pH 11.7). Chcmilumincccncc intensity was measured using a Aloka luminescence reader (Aloka Co. Japan) (waiting time. 10 s: integration lime. 10 s). Buffer salt in SO 2» Specificity lxlO^mol/I- CL(%) 1 Na.S 100 L-Cystet»'" 201 t.liil.i[hi..n 41 1 IMihielbmlol ŠJ0 1 WHOi 1 7J> M \nn 34 Vitamin E 2 ' SOtsocn IJ 1 •ut<»«wiiw>1™«d«l( 1 -ofttmiuj Metal ion Effect by Catalase&.SOD on CL intensity 1 Cu*- CUT W Iftdrogen sulfWfc crwilml tit [Discussion] * This is a novel chcmiluminescence method based on the principle that light i sulfide in the preset emitted by metal ions and hydrogen • The effects or sever il mcial ions (copper (II). copper (I), d aluminum) were studied. Intense eneralcd wiih copper (11). | ' Reactive oxygen in olvcd in this chemiluminescent reacnon 1 wti analyzed using 1 SK by the addition of a scavenging enzyme, SOD. Luc itenin was essential fm ihc generation or ^ reactive oxygen, W il. the addition of hoih cam lose and 1 SOD. this signal cs. lOMhdly disappeared. Tills result 1 indicates ihat the rn dienl species is a superoxide anion The 1 " Fi8.l. • DcicciionlimitofNa.^»zis20p^/ast«y. • Reproducibility was from 1.5 lo 11.7* tin Imvn Iwn uviilmMwl to In> ...ii , Willi N nvailnl*! nMMtnnmnlnl tpaOthVM low niinigy '«i>l)(»M, Itw oo»t tl.wmlr,il slon woukl tx> n UnntiiHHil liv tttn Ryillwru Hid MONMM o^Mtnttnim WftOM MMIMJXWRnfl i»»k potitton* mn KHnlml nl nlnjlitlv hMjIvni photon miing-nti (linn llvn fiml Molfltiwl |wnK» A flint , rtnopki* tinnlionot ol Inn (too nlthHon sl.itn* iv tlm KyiUwiu innlntM'ii m nn HittHnMiii,) topii' <<«|i«H,inlly (torn tfw> vhiwi*»iiI ol txtKlK »ot (ht|N>ndnncy l>|n shk«iw mm j en,» i-h#Mi a tin 11MI10.11 FurpoH "......linn-. Wm»H)rfj|iy Myology - *•*«<«»< "»>»* •"*"* ■ B3LYP nuo-cc-pVTZ IAO) HOMocJJ»*o'o«e- MtKhodolocy - cn..p.nlcl. °W ^ar«ie " ......' Greens hrcbori Results - basis set dependency - toss (PW-AO) »nC «u9e ibto N*»*en at -■ H......1I-. Ph«»rt«oipllo« .1- Current ecological knowledge about Mus spicilegus Petényi, 1882 (Rodentia) in Slovakia Csanády A.1, Stanko M.2 3 & Mošanský L.2, 1 University of Prešov, Faculty of Humanities and Natural Sciences, Department of Biology 2 Institute of Parasitology, Slovak Academy of Science, SK-04001 Košice, Slovakia INTRODUCTION 3 Institute of Zoology, Slovak Academy of Science, SK-040 01 Kosice, Slovakia > ecological research in the years 2002 - 2010 from four orographic units of Slovakia (Východoslovenská rovina plam, Košická kotlina basin, Ipeľská pahorkatina upland and Hronská pahorkatina upland). r- focus on autumn-winter and spring-summer ecology, reproduction, mounds morphology and morphometric analyses RESULTS r the variability of the overwintering mounds (n = 376) and the nest size (n = 83) were confirmed between the orographic regions and between the habitat type (Csanády et al. 2019, Acta Zoo/. Acad. Sci. Hung. 65); A r- winter prevalence of juveniles (x2 = 52 74, p <0 01) and males were confirmed for the autumn-winter period (x2 = 5.47, p <0.05) (Čanády et al. 2009, Acta Zool. Bulg. 61(1)), B > spring-summer sex ratio was in favor of males (x2=1 96, df = 1, p>0 05) (Csanády etal. in press, Biológia); C y we confirmed higher testicular values in M spicilegus males than in M musculus males (Csanády et al. 2019, Mamm. Res. 64); D r average litter size (n = 9) was 8 3 (6-10) embryos per female (Csanády et al. in press, Biológia); C > four dental variables (LaM1, LaM,, LM, and LOID) suitable for the differentiation two Mus species (Csanády etal. 2018, Fool. Zool., 67(3-4)), E rWI spicilegus population significant morphologically differ by longer head-and-body length and shorter of tail length from M. musculus (Čanády et al. 2008, VOCS VIII., 2007) CONCLUSION Long-term research significantly expanded our knowledge about morphology and ecology of M. spicilegus at the margin of the species distribution ACKNOWLEDGEMENTS We would like to express our sincere thanks to all colleagues for their help in both the field and laboratory work. Research was supported by the VEGA projec^1/0084/18 and 1/0277/19 PAKri Aktivita klíšťat u Brněnské přehrady v minulých letech H. Nejezchlebovi, A. Žákovská, V. Nesnídalová, K. Bečárová, B. Kolářová, R. Horáková Oddělení fyziologie živočichů a imunologie, PřF MUNI, Kotlářská 2, 611 37 Brno Cílem příspěvku je podat přehled o aktivitě klíštěte obecného (Ixodes ricinus) v průběhu minulých let na lokalitě Ruda (234 m. n. m., 49°14'18" s. š., 16°31'29" v. d.). Lokalita se nachází asi 200 m na západ od Brněnské I přehrady. Jde o oblast navštěvovanou turisty i místními lidmi, kteří zde sportují a tráví volný čas (obr 1). Sběry klíšťat probíhaly intermitentně v letech 2012, 2015, 2016 a 2019 v pravidelných intervalech jednou za 14 dní metodou vlajkování od března do října / listopadu (obr. 2). Výsledky jsou uvedeny v tab. 1. Obr. 1: Brněnská přehrada a okolí: oblíbené místo k rekreaci celkem larvy nymfy samice samci kritický závislost na měsíc teplotě vlhkosti vzduchu tlaku vzduchu 2012 321 64 236 7 14 červen ne ne ne 2015 255 12 232 6 5 květen ne ne ne 2016 364 77 261 16 10 duben ano ne - 2019 554 134 411 2 7 květen ne ano ano Tab 1: Výsledky sběrů v jednotlivých letech odpořeno ze Specifického výzkumu MUNI /A/1397/2019 Obr. 2: Vlajkování na lokalitě Ruda Roční úhrny nasbíraných klíšťat značně kolísaly a statisticky se od sebe liší. Nejvyšší aktivita klíšťat byla vždy zaznamenána v jarních měsících, s tím je spojena i míra rizika přisátí klíštěte na člověka/zvíře. Nejpočetnějším stádiem byly nymfy, průměrně tvořily 76 % odchycených jedinců. Nejméně zastoupení byli dospělci (5 %). 19 % odchycených klíšťat tvořily larvy. I zde nacházíme statisticky významné rozdíly mezi jednotlivými lety. Závislost celkového počtu odchycených jedinců na teplotě, relativní vlhkosti vzduchu a atmosférickém tlaku nebyla ve většině případů statisticky potvrzena. r llosi response to Borrelia afzelii in BALB/c mice Department od ('om/itiralive Animal Physiology and General Zoology, Faculty of Science, Masaryk University, Kotldrskd 2, CZ-61137 Brno, Czech Republic: tel.: +4 420 5 41129396, email: * > ltd J? M ill mil i\p M>TIHHn P«-«.J l>«Uf—» M|IM |W tm ttm wtmvimtt t ** MM M j *, '•feWfJUlffci fee; •4 MfrfiWB«riii1H,iu&f > mm m**4i bwt mm jr«a ■ i ■ J f.TMti. •.,111.1-,,:., »MJ«I MJMIAV —till 6-year's study of the presence of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks in the town-park locality Brno, Czech Republic using DFM, PCR, PCR-RFLP and PAGE methods (Ml Nejedlá Petra, Mejzliková Marie, Žákovská Alena, Holíková Alena. Martiniková Hana, Justinova Lenka apartment Of Comparative Anirnoi PhywQfogy end General Zoology. Faculty of Science, Masaryk UntvOraily. Brno. Czech RopuWtc Introduction Tt* motets <*vm *******itew.ljMtw^ lorm« ec*iVaxßwrw^ 9000190013* In Europe fh» gnfmpaoat (8 buwivfori wnutWOo, D oarvtf.fl «• ••• [i řMBťWř*ifunni int»,3 si*** B 6f) mmáuiběauMdbttMtohmm RM UrttiXow iwyMH fcmarlk* mB*i-*»t,and MMr/of imm rXnurj 6c-U *>» oNa pwvnc* ry ^íoogcrv: Som>*j» tx*&X*U>« vjtou l«to «ino ujAn-part «acardy Birx«. Ooch Ropubhc Materials and Methods to !ho yaar» o* 1996 - 2002 * toiaí of 3170 lioOn txwixrt fxfci hvi ccfloood by dragging d *M« flanneJ ctotn ow b* vptUton Tho Ida tm» «xamnad to* bSe p«««* of spľocf-Ttu by d^-f-efcJ ffKTosoopy (OřVi. umpkM arten moro man W weocne*« wore franalorrod arto BSK+t medvn and iiMiom*] under íTC for tht 0**oA4n of ap*od*ito» *w« u*so Mo PCR • from the Ik* rrwjgu» harboured mor* man 100 KWocnetM rwo wnpNi (on* tor PCR on) v>cooo for tit* cuKvafeon m bsx'H meourn) mt« proparod In 2001 onfy PCR waa mod. ai J002 trvw typat of meíMOi wo iMd (PCR. DFM and eutxaton «wen towing Of U and PCR) Al ma moroott n«m tor mod 10 obtain x«rrxivta ttttnm !** luriht* dtuafed 4*H and waR»d) Locality >>•.■!'•. nanus H kKNffy ß/ryj C/o lha town. 107-2 i tody okA in a uUvatMyi In uva Orwrcia mpur. < » Byt,ai*.u Onof pcorfi/ta Pppcrfua irw>\A3 ftobna n poftgitftt Lofír áocrímt mo rxwaenl ahfub kayar « corrpoaa-í pwainry of id fcm ivaowwi 5*n6uív» racavnoaa, nvuUrra SwŕA aonpuneji wvl i^ndafjrtv^ŕi conlawn ^Avn unmOrri. utm rama. Awruna rarawavdaa, Ala. A50 nffflóreAa *toiiunpjt\m foul tjf0*' (f nJN PrUJ '.1. p P r* vN Pr> ) pcr t ••• ■ , ,■ ■ ■ ' ■ MU Pi t 1ST» l&S 1999 MOl 2O01 1 eucuo'r) XCOHOti) MHÍ7JI 1 oorODi 440110 0 i 3/13 031. 1 ■ ■ jal 6 r, 7J7Q (100) 10/l!7p9) 00(001 2012 7) 3.v. f- ;, "ansa o &j 31r7ř9(l1 1) 13/106 p 7) ■., ji (iv (13 4) ■ Í/WS5I au (00/ rvj too) 1/73(14) 1/76 (1 3) 1/147 (SŤ) •m./ - í. »3/4« |9 71 57/473 (17 3) 1 :/•»/.■ ill., 37/797 (4 0) !!/! A ' 1 c j »1257 (11 7) 1 ■)/•«) C/U) 17/797 (7 1) 1!«l ftl) «1/797 64) '.'il ľ 4. « 4a{ 4JJ 350 tfÄ . 1 i/ltl . /j ■ ... 1 ■■: < ■■ . ■: . ' i Jl/U iBmo OatfiRwKK »« WWW «| Mt I IM nul iwnoM/Wku 72 %(m*n lin, IimNi 82% irrxnDb'* tv»» 7 7 s, tt» poh 'M UJ 9m W pa<*W, 04 a^lQC4^^ »Hl— »fl íl l^llu tfy h*fc <4W| 70 '-^'*"J loa I" 7 left ''/ii^Wn Mrak^ntlbtM a4uj^j t/, niimwi A 10 K. UM -w. ■ [Tni M ( Acknowlodgement »»7i 7 1*7 43 V7«4. »Z7 N mr " »4~» U*/4 ■V, 7 0 <7/^ dpt.; N 1ST» 0 94MW BXľi M 1147(1 B <4|«* m: 1« u xai 77 ^74* BM7Í7, u Ma 73 <7 1} m p au Wi ř3 * * Isolation and characterization of a novel subspecies ot Photorhabdus asymbiotica isolated from Heterorhabditis indica Ryuset Kuwata, Toyoshl Yoshiga, Mutsuhiro Yoshida* and Eizo Kondo Department of Applied Biological Sciences, Saga University, Honjo 1, Saga 840-8502, Japan. 'National Agricultural Research Center, 3-1-1 Kannondai. Tsukuba, Ibaraki 305-8666, Japan. Introduction Photorhabdus luminescent and P. temperata form a mulualisttc symbiosis with heterorhabditld entomopathogenic nematodes. On the other hand. P. asymbiotica is a human pathogenic bacterium that has been isolated only from human clinical specimens in the USA and Australia. There is no assertive information available on the origin and the natural habitat of this clinical species. In the study of Photorhabdus bacteria isolated from Japan, we found that two bacterial isotates from H. indica show high similarity to the clinical isolates of P. asymbiotica. Here we show the phylogenetlc relationships of the Photorhabdus bacteria and H. indica associated with the bacteria. To investigate if the H. indica isolates are able to form mutualislic relationship with clinical isolates of P. asymbiotfco, we monoxenicatly cultured Japanese H. Indica isolates with some P. asymbiotica on the nutrient lipid agar plates. Pathogenicity of Us obtained from the monoxenlc culture was also examined. Results Onlr-10 OnKn2 Growth and reproduction ol H. indica on different Photorhabdus isolates. HM1 , «II.I mi Phytosencttc relationships amons Heterorhabditis Isolates based on the sequences of COI region. Phylogenetlc relationships among Photorhabdus isolates based on the 16S rONA sequences. Habanft Meg HW79 XILIt ^ Telgjin I HbT 'Hm HMUt MVI6 «80 |r 980W4I, " ' '- CCHOOI I CbK)16J P Onlr-10 J26VS6' 1216« 261/07 P. temperata P. luminescens mi r P. osymbfot/co (Aus.) P. osymbrotrfco (USA| Phylogenetlc relationships among Photorhabdus isolates based on the gyraso subuntt B gene sequences Conclusion % ssrsubspctie5 o( p- °mtk° XXr°r "1°/'? WHh ^°™P»nlc nematodes S tSnSiStS&S'S^-,5 CbKj16.1 and OnlrdO with those ol (mown Photorhatxlus species .....■■"i.'U'IOI I ' rni wii«i!iiin U Phatorlialxtm sp. ChK)l6.1: 2. Phatarttatxiu* sp. 0nli4U; .1, »"«•wm-TmÄ.,! '* t>f^ SutKoccwolla tni Phvfodocta 2 Th, mwgo of Subcoce/rwto no t. mo»i «h^ľ.ľ*bl* 10 ,nva»,on the nematode H baetmtopharn '■ 6 IWTIM I» moreoatoaZZ.to./ '"vaded bV m» 5poa.« H oectenopňore Ftg 7 Infected Imago of Subcocanlla Installation of a Swiss Zooparasitic Nematode Bank S. Kuske, J. Gründer, E. Fischer University of Applied Sciences Wadenswil HSW. Grunlul PO Box 335, CH-8320 Wodonswil. Switzerland Email; s ktiskodplisw vh HSW Background Worldwide Ihere are several nematode families known expressing bio-conlrol potential, with a broad range of nematode) products available (or commercial use. In recent years, more than 40 entomopathogenic nematode strains, belonging to more lhan 10 different species from more thanlOO monitoring siles. as well as aboul 100 isolates of the symbiotic bacteria have been collected and reared in Switzerland and are currently held m calibration at Ihe Swiss Federal Research Station Agroscope ACW Wadenswil These nematodes and their symbionts are available for further investigation concerning their bio-control potential Objectives The goal of Ihe Swiss zooparasite nematode bank is to preserve the genetic resources of endemic nematodes and their symbiotic bacteria. II aims to ensure quick and reliable access to both nematodes and bacteria for research topics at the University or Applied Science in Wadenswil (HSW) It also alms to make sure lhat the biological activity of this living organisms can be maintained and stored using qualily-ensured standard methods The Swiss zooparasitic nematode bank will provide an universally available standard which can be used as a base for tho development of new commercial products. Species list EPN's Heterorhobditids H. bectoriophoro H mogidis Stolnernemalids S ollimi S. Ohsnanum S bicornutum S. carpocapsao S lotliao S Swiss 'gliisvn' typo S mlmimtltum S kraussQi S. WQISQTI Baclorln P mii:,i..i.. ....... XimtiHMtxius specie* isolated from Ihe Stotnomima spocios listed obovo nematodes bacteria I- identification -1 rearing/cultivation EPN'r, . ll.ll.H Ii lľ .ltl..H Blotosut: > virulence ' host i.inai' r peisistencfl/longevily r lomp/soil/moisluro preferences Evaluation of be potential lor commercial use HOCHSCHULE WADENSWIL Methods The general methods used for cryopreservation are based on Ihe recommendations of Bar ol al (2004) > Fillration of 100 ml IJ through a Whatman Nr 1 Filler. > Immersion of Ihe Us in in glycerol (18% for Slemornemo spp. and 13% for Haterorhabditis spp.) lot a set time (48 hours for Sleiimmoma spp. 168 hours for Hotewrlmbditis spp.) The number of Us was ad|usted lo about 12000 Us per 1 ml glycerol solution > Vacuum filtration onto a filter paper disk r Immersion of Ihe filler-paper in pre-chilled (10'C) Methanol 170%). > Placement of the rolled filter-paper into pre-chilled (0"C) cryogenic vials and immediate immersion of the closed vial Into liquid nitrogen The nematodes are thawed by pouring 1 5ml of Ringer's solution (24'C) into Ihe cryogenic vial Virulence of selected strains was tested against the following pests: > Black vine weevil (Oliorhynchus stikMus) r Hazelnut weevil (Balaninus nucum) > Chestnut weevil (Ctmulio oloplias) > Chestnut tortnx (Cydia splendana) > Cockchafer (Malolontlia motolontha) > European cherry (mil fly (Rhagokttis cerasi) > Walnut husk-fly (Rhagoletis complola) Perspectives y live databank encompassing many species > quick access > good quolity > new species/straws <• ongoing characterisation > new products COST650 (compMod 2008) COST862 (slnriod ZOOS) Dvel mitochondrial gene arrangement among some species of the Genus Heterorhabdith Sassia (). Rggcaj and Ann M. Burncll of Bioengirteering andAgroecofogys National University of Ireland Maynooth, Maynooth, Co, KUdart\ Ireland Irsrloul (few pi Hi-f nHl>V\ mMnim rnxn // ^»'«* r«TO*Jmii DSV M I (I HhpK M**llV**1»».NOH i«crtui»ctjrv //. ' ,. I. rl ."'i I» «*fcuHl*roJ hi*.C*r*\ • * tramtrjnlipn »I * Hoc* of wven tjchev iR* A lj. i* • MthtvmtarftfM iftKA4 «na Mr«ic»«rui • a lramk**tpirt r»l iK Ni*, I > wd iRNA * ■ UMAI.U'l^lKMll.iRKA l.uJKtM HiKioiimMiiii '-'Trt-jf-rr wi»ui rirtl«l*«l»llimcm . :.-.w'.-MW .. *I«(,Mui I ■y->M*itTriiT^?«rtlmliw!mi ». i i»i «•-• ■»« «.; ■ vi IU reo ml 1* rrlcJklwil* ulntJJi__________, ■ r>rtl__ JwrloUH_____________. , __ , *5_!"I «••"' COST COSH tempts to Develop an RNAi Protocol Nil I MAY! for Steinernema carpocf I. Din. I. Tyson and A.M. Br Department of Biology' -variorial University of.lreland May. Email: ilona.dix'ii luay.ic n traduction ihetf biologic*! (bTxtmr m*) k k.J ininc-lt nftwiri inlcnMcopk 1111.11101 t.ilMI'l wwiiM* tvrjwyfM ....... ' Lil u,pr„|„( in RSAi dim t.HilJ uai K' < •..Aim, .„«m> <„ ,|,HN,\ ..I.., n.M iwcwilnll i» bj inj.vlmji d.RNA nf IOWA <>t IK- utmi ||i,B|> rt*u*i l.w Ihi* In Ihjl Sttimmrmi (MifttirsviwV 1» .•I itMmw) f.«t tlx RNAI nrakinMl Aktmnn/h du i«i..i RN \. ■ ly/f^.V'tu* nvjfj MhMUm Hhith li\er> Jill'crml from tkai ,>f* .V,-^..- in. * 1'»' *v»>ii.il MiiHliin ill mi .■■IN......... ilv ii "*"*—•^J"'n nnminnii ttj'iM in n nlii m nriiVt ifltii >■ .h.™i. t>> Uw fan i »»»0 oriMiu iKicTiiiop.u.t-nKi wc r«vivi> Reference* i I In, A«lflf Kmkh .nu.fHv.iv ipncik lt.ti.ivitf.K.t., j....hi. feinmM kv\ .....«'-•/•..'•./.....'....„, '...,„,. »nr.nn ,|.,u«, ?.i«». A.I..I.J fiwitiMmimnDjit-aijahaft ,-*^r,,.t», '" '■' .....* »N \ "M.n. i, Voflm- MM I III,;,.,,. \, i.ledgctncnCs Ptltti'PrntntMl m ifei i i < oil |$a-n,,,,.........m„,i...... VS in k>llii|t, 11, i, f Minn i , I ft.hill. JIHIO. Testing the Male Colonisation Hypothesis in Steinernema Species rj Mohamod Alsalynh and Chrlstino Griffin NUI MAYNOOTII Inslltuto of Bloonginooring and Agroecology. NUI Maynooth. Ireland. Introduction: According to the male colonisation hypothesis, infective juveniles that are destined to develop in to males invade insect earlier than females. This reported to be the case In several species of Steinernema by Grewal et al.(1993) but the hypothesis has been controversial. Stuart et al (1998) tested the hypothesis for Steinernema glasen. one of the species reported by Grewal et al . to show male colonisation. If males colonise first, the sex ratio is expected to male-biased following a short exposure to a host, but the bias should decrease with further exposure. Insect were exposed to S. glason infective juveniles in sand columns for various periods of time, but there was no evidence of earlier colonizing by males than females (Stuart et al 1998) In this study using the same approach as Stuart et al we ask whether males or (females) colonise first, by looking for changes in the nematode sex ratio in hosts which are exposed to infective juveniles for different period of time Since lime of emergence from the host may also affect sex ratio in steinernematids (Lewis & Gaugler, 1994), infective juveniles were harvested al three times and harvests were tested separately. Results: Methods: Culture and harvest of nematodes: Steinernema feluoo (4CFMO strain) and Steinernema carpocapsoo (M 8,f(Bn, ^ rea[ed |n Qmm m.„onl!llo ^ harvests were collected, from early (Day 1). m,ddle (Day 6) and late (Day 10) in tho emergence period Assay: 6 mellonolta larvae were exposed to infective luvonHes (100/insecti in sand for 4. 6. 8. 10. 20. 24 and 48 hours and washed A 4 cm sand column was used for S toltme and 1 cm lor S caipocapsae After about S days the Gallena were dissected and (he number of male and female nematodes counted There were 10 replicates per exposure period ---Nematcodfl Plastic pot Muln-well plate Fig. 1. The number of nematodes invading wax moth larvae increased with lime of exposure. More female than males nematodes were found at each exposure period. S Initial) Early harvesl S caipocopsao 1 • * id n m *t Eatty rlitrvMI ill narvou * • i If) JO Jí il I • I ■• ■ ■ ill. f „vi-, fhf — Mfl 4 * « ID R N I * * • w » h «a • .....torn mi Flfl. 3 The proportion of mules wot, higher in lalor harvtKis Inr h«k lire proportion of male* In .ho harvest „rne. being higher ,n nematode, KK2K*teSil2 nalal cadaver S tolttlKl nomatodos that emerged later from tho v on P-001Q OJS «« 3 lala I ©ally to? ■ 0 /r,„....,r I I I BWly rnldalo Hun-ant Fig. 2. There was no systematic change In sex ralio with time of exposure for any harvest of either S to/l/ae oFS carpocepsve S foltiae S carpocapsoe Ea/ty w.,i+i 111111 : ■ 111111 i os OS 04 mini •mill a t n «,t }> i, w 1 For S ctirpocopsati. there was a Irend " for the proportion of males to increase ** over tho 4-6-8 hours period . but Ihoro "* was no significant difference in «ox •» rallo between those exposuro limos » when Ihe experiment was repeated (data not shown) I I I I I I f Conclusion: 1 There was no systematic chongo in sex ratio with oxposuro lime for eiihe. S (ottino (previously found not lo be male first coloniser species) or S ca,pocaps«o (reported by Grewal et al 1993) to be species with mato-lin.! coloniser) Therefor, our data do not support tho male cotorwation hypothesis for S carpocnpsae 2. S cmpocopsao and Stoltlae are female biased In .he sox ratio 3. Male tend lo emoifjo from Ihe hosts Idler than females ,n both S.cutpoaipsiw umlStattmo We are currently leslino other species of Stoutamma in a similar manner. Rotoroncos: I On»i V V, SWvan ». I**. EE «1 0.uv»r, « l»J Mak- ~o"W- ""WW** • eokwai'ngaw J r«m»nlu MOI SOJOOII i Una I fi aikicuugui B llHM LMommalnotHiiw namalodaa iHnaema siaawnamawaal aai raw n>l«l»i til (maging Mulagy J l«wl«»»l« PallMKW « MS-MJ J Hou. J am\ AM nat* Oausw. K '<™ a.' "» •^*T1^T,^T " ^trn.K^il.ogwicnamatam «wMWMamiMgiai J »<«t*ial.falN*e» « »*•»» A report on the occurrence of Heterorhabditis indica with two symbiotic bacterial species in Japan M. Yoshida". R. Kuwata-T. Yoshiga-" and T. Mizukubo" 11 N;iiiun:il Agricultural Research Center, Tsukuba, Ibar.iki 305-8666, Japan 2) Saga IJimcrsity, Saga 840-8502, Japan , W Body colour of cadavers of Galleria metlonella infecfed by Heterorhabditis indica Cadaver colour; 19 RH type N-RH type Fslw96 TkOgS, 10 W K>Yk«87f7l/-r- Distribution of heterorhabditids in Japan (1990 ~r» u a ss-s ia » »—»—n a u u—3T" ■ : Mortality of Galleria larvae by H. indica infection at 5 days (20~35°C) and 10 days (15°C) after treatment; • :First day emergence was observed; I Galleria / 10 Us in 24 well plate, 8 wells / replicate, each isolate 3 replicate Reproduction activity ends with Pathogenicity and reproductive lost ol reddish brown colour activity is retained n symbiotic bacteria 4J- were examined ,Q- Photorhabdus luminescens New subspecies of, P. asymbiotica &/'. asymbiotica : clinical isolates / relationship with EPNs unknown ©about 5~ 10 years after isolation, the symbionts were studied Questions: ~Ei 1. Was P. asymbiotica a svmbiont of the N-Kli type? 2. Did replacements of symbionts o^ur during laboratory maintenance? Symhionis ill the isolates obtained from Galleria trap have la he examined. A survey for //. indica in some regions, where the N-RB type has been isolated, is required. Survey for EPNs in southern Chiba Prcf. (Cb), 2005. Sampling location (f) and positive sites (arrows) for Heterorhabditis indica. Arrow's colour indicates RH and/or N-RB lypcs as follows. ■ iRHiypc: j: N-RH t> pe: ■■]: Rl) and N-RH type Habitat characteristics of sampling silos in the southern Chiba Prof. (Chi (9 locations. 21 sites, 120 samples) —Distance N<> l»"'avi- maaS&iis tin lioiinunl of Hit' local ions whore I ho occurrence ol'//. indica willi h\o symbiotic bacterial species wore observed. Conclusion @ The follow!ngS were proved by the Held survey. 1. //. indica has ti symbiotic relationship with P. asymbiotica too. 2. Insects infected by //. Indica with P. asymbttlca have a dark grey u> pale yellow colour (not reddish colbui)., ui is there any environmental isolation (hairier) between die hyp symbionl types'.' 1, Habitat m, Woodland: I1) sites, asymbiotica type from 1 site (Unir-iU) Grassland: 8 sites asymbiotica typo from <* sites (all -I sites in the south part of China Pwl Boso Po.....SUlS I 2. Two eases mm the adjacent occurrences asymbiotica typo: sou-side « lumineseens type: inland-aloe ^ =» Did asymbiotica type invade alter luminescens type colonjxcd t Where did asymbiotica type come from? DW II WiBHJ......' «** 1, I lost preferences: further research required.___ A review on entomopathogenic nematodes in Turkey Alper Susurluk Turkish Ministry of Agriculture and Rural Affairs, Plant Quarantine Service, Department ofNematology, Liman Str. No: 11, 35230 Alsancak /Izmir-Turkey In Turin. coMnceuihocenic nematode) (EPNs) arc considered as potential enbpcal control agent against soil-bome insect pests. Thus, there are some audits on EFN's in Turkcs a« in .ill over the world. The aim of the PHcniiM is to cavil) the situation ol 1 PSs's in TunVe>. The first scientific opmmeM u Turtes on EPN> earned out b> Otter el a! in 1W5. Many of the* studies consist of nint} experiment* including extraction and (jnemtitid and one heurorhahdilid species •e>>. In ibesc suncss. the most common JJition lo common species, a new species t by lluk et al. (2003.). Mans of (he * mofcculai technique. PCR.RfLP. All to using of GaOttto mltonellu'bail que*, fist ster end of the IpKies »»> SWatnMM j&kx |„ a** beta described as X moo/ic, ascribed" species »ere eatneted "KttN* Besiiet Hie Qwtt ^ ^l^r ™h « «mefc«J studies on *<" are no ovals- cxcetime'i' '. P*?nlul 01 d'ficrcnt temperatures But ^^^^Sf^t^^!^^ " 0nlv l"° M, because of lacking 1» MKtuMoB, moKix, -U. ,k T .. 7^h^(ini.kw "t'ponun.iics f„f conduetin. J 1 u"u'o/i>nj,. At present, all described EPN species in Turkey Nematode species_Isolation regions StcSntrnema fihiae Authors £ anaioiiensi S. eprpffstpSM 8 affint UeitrarhabdiUs Ankara. Kirschir. V. skisehir (Center Anatolia). Rize and Sinop (Nonh of Turkey), Burdur (Sowh of Turkey), Van (East of Tttrkey). Canakkale (West of Turkey). Kars (fast Anatolia) Anialya and Ice I (Soth of Turkey) feel, Adana (South of Turkey). Mardin (Southeast Anatolia) TokanNonhofTurkev) lckirdug,Kirk]arcli (Nortjv^tem Turkey) Ankam (Center Anatolia) ••\»Kara.Aksaniv(Ctfn A|wioha). |sU)nbu| Kirkiaroli, Ickirdag Wordnvtsten, lurk J^^cntcr Anatolia) O/.er et al. (1995). Susurluk et al. (2001). Ha/ir et al.. (2003b) Hu^ir cl a!., (2003a) Kepenekcj (20O2) Hazir et al. (2003b) unpublished Susurluk et „) (2001). Ila/ir « al. (20031,) 0» c. • vr POSTER 13 I Christian-Albrechts-University Kiel I Faculty of Agriculture and Food I Science Institute for Phytopathology | Dept. Biotechnology & Biol. Control Checking for synergistic effects between Bacillus thuringienis and Steinernema carpocapsae against Plutella xylostella introduction: Tho Diamondback Molh (D&M) Pluletln xylostella is a mapr po&t of cructlers Duo to the intons/vo use 01 broad-spectrum chemical insecticides the potential of invertebrate natural enemies Is limited and tho post has developed resistance again:! al chemical insecticide, even agalnsl the biological Insecticide Bacillus llxjringienais (Bl), In order to reduce the tosislanco level alternative biocontrol agents ate needed lor DBM control Enlomopalhogenlc nematodes (EPN) wore suceossWy used against DBM and alternating applications wllh Bacillus trwingiensts resulted in good conirot during field exporiemnts on Java. Indonesia. The objective ol this study was 10 investigate whether synergistic offects occur if both agents are used together airbrush nozzle Material and Methods: EPN: S. carpocapsae (EN03) o noma GmbH. Germany Bl: Dipól ES* 1B1 var. kurslaki) Stabler Agrochemio GmbH S Co. KG. Germany XenTarl" (Bt vat auawah Valent BioSciencos Corporation. USA BlomQk (Bt var, israoleruls) Blola. Germany Surtaelanl-polymer-lorinulatlon (SPF): 0.3% Himulgan and 0 3% Xanthan Application ol EPN and Bl on cabbage leal disks: EPN: 40ml wilh flat-Ion nozjlo Bl: 1ml with airbrush nozzla Fig. 1 Bl-Applicalion One leal disk with ono 3" larva In oach well was incubated 48h at Z5"C and 80% RH. Fig 2 celhvoil ptoto with cubbago leal disks Stall slic analysis: P,-t-(1-P,)(!-P,)<1-P,| enpocled monnlity lor additive ottocis , P.. P, mortality at single component untie analysis ol dittoroncos 10 additive ellocts worn cariiod out with Fishors oxocl Ifshintll Ciri,0rtB '0r *° ,r,,eroctlon (according 1o Finney, 1971) ■. Il..1» 1. 1 D..O. 1 synergism not -.iui.iff. oni D..D, addlllve ollect *—i———-..........it ui uooa i.iiviH ( D„: obMrvod numb«f ol doad Inrvao Xiaoil VI and RaH-Udo Ehlois institute lor PhytOfMlhology Oopi Bloloclwol t Biol. Control Mii'iniinn-nadawau.si, g 24116 Klol Tol .49<3i8aoa864 Acknowl Result: íi1 :;! ii ■ n n« II Hi.lli Bla.lti Fig. 3. Effect ol singlo and combinalion use against P. xylostella 3" Instar larvao 8lk:Dipel. Bla XenTan. BliBlomOk. concentrsllon: 20 ng cmn-3x24 PI k n Í1 n D • t»N ■ '••tl'N Fig. 4, Effocl ol singlo and combination use agamsl P. xylostella 3" Inslar larvao Btk Dipol, Bia: XenTan. EPN: S. carpocapsae EN03. SPF: 0.3% Rimulgan *0.3%Xanlhon, concomrallon 20ngcin'of3-5U&riarva. n.3x24 Tab. 2: Interaction ol combined agents against 3" Instar P. xylostella Interaction combination rsp«-eaten ados-uva synor-o.-.ťc tni ige ™»t«: •pray method Oipol (zOngcm") . B/omůk (20ixj.'cm'| S 8 rnivod XonTon (Mne/cm3! » BiCffluk(20ng/cnV) 8 8 Dipol IJOno'crrO . Xeii'liiri (20ix>'cm-'j 8 4 1 innod Oipol tzOngvcmr) ♦ XonTari izena/cm*) • BJomuk (řono/cin-1) 4 3 1 mined D.00I (JOno-'cm-i. EN03 tS/lnivíil 4 3 1 ono altui anothei XonTan (20iv'cni'l . EM» lilaiva) 4 4 ono artoi aiwtnof SPF . ĚN03 lllarvill 4 3 1 rniAiiťl _ EPF . XenTan tlOrnO'll 4 3 1 m.iod XoiiTnnllOdxHI . END3 (ilarvu) 4 a 1 nnice SPF . EN03 (jvtarvo) . XonTart (10irx>1| 4 3 1 mied Conclusion and Discussion: 611 has no toule etlocl agalnst DBM rlio.ílly Oddilive ollocta locordod with cainbuuilions ol Bl (Btk. Otal and EPN 01 Ulk, Bia .mi 1 Illl mm od applicotion ol 81 and EPN la oconomicolly not feasible niloniating appl«ationii ol Bl and EPN are a poworUil and lokobio lom 10 avoid or relord dovolopmoiil ol Bt icsltlanco In P. >ytontu«a lllk .111.1 II' 1 -.1,1.1,1.1.............■ iilleiiHitiiiil auiJli .Iťon:. Pm|iii:l! t-U ICA4 CT-20OI 10003 OIABOLO Art itilogrnlh/e tjltatugy lor lha suslalnablo conlru Ol rlninioiKÍb:u:k rnoih (PlitloHo xyioilolla) by conovrvulion ol iiiilurul onomiou und npplicalioii 01 bloconlrol auonlK An experience on biocontrol of Catiroa varipes (Klua) (Hymenoptera, Tenthredinidae) by means of EPNs in an urban park of Bologna suburbs (Emilia-Romagna, Northern Italy) Giovanna Curto, Nicoletta Vai Plant Protection Service, Regione Emilia-Romagna via Saliceto 81, 40128 Bologna, Italy gcurtaQbregione.fimilia-romagna.il. Caliron varipes (Klug) Hymonoplora Tunthrodinldno at Italian latitudes complolcs two generations per year and overwinters as mature larva eocooned In the soil. The adults, coming from overwintering larvae, fly from April to May. The females Insert more than ten eggs under the lower loaf epidermis. The larvae hatch at mid May and develop until the end of the same month: when thoy are mature drop and bury thomsolvos Into the soil, where thoy cocoon and pupate, giving a second generation at the ond of Juno. The larvae llvo gregarious on the leaf lower epidermis, eating tho parenchyma as long as thoy skoletonlso tho leaf, leaving one cutlclo intact The C. varlpos control Is based on culling and removing tho Infested leaves, but in case of heavy Infostations the cutting could be very drastic, and only biocontrol molhods can bo accepted by the frequenters of .in urban park. lontr^ n Pro,flcJlon ^rvlce experts were asked for contro ling a very heavy C. varipes Infestation In a small oak wood In tho centre of Budrlo, a small town In province of Bologna, Northern Italy. OBJECTIVES To check the effectiveness of: - A soil application of Heterorhabditls ba^r7ophoS*irMho control of eocooned larvae before their pupation. - Steinernema teltlae, formulated for foliar application, sprayed on the gregarious larvae on Infested leaves. MATERIAL AND METHODS S. feltiae application to the leaves On May 24,h, 2005, 100 oak leaves, heavily infested by 104 larvae of C. varipes, were collected from the oak wood; SO leaves with 54 larvae on top, were sprayed with S. faltlae, the other ones with 50 larvae on top, were hold as untreated control (table 1). Assessments The leaves were maintained wet at 23 C; a week later, the larval mortality was assessed, the larvae were scored as dead/live and a sample of the dead ones was dissected and examined under a stereomicroscope for checking nematodes inside. Table 1 - Foliar application. H. bacteriophora to the soil application Beneath the most damaged oak trees 100 m2 were bounded and Irrigated; H. bacteriophora was applied by means of a watering can, during the drop of the most mature larvae (table 2); then both the treated soil and 50 ma of an untreated area were covered with a net. Table 2 - Soil application. Active Inert......it Comm. product Dose tu. i,.,') Application time 1 Hclctothubctltl* bňctcitophotii Ncm.flop 5x10' My 31". 2005 ■1 Untieated control Active ingredient Comm. product Dose [IJ.Iin'l Application time 1 Steincmcnn telttae Ncnnsys F 5 x 10' M,iy24'". 2005 i Untreated control i At tho same time, 40 mature larvae were collected and transferred to the nematology laboratory, where they dug and buried themselves Into the soil of two pots, each 16 cm diameter, before applying H. bacteriophora in the first one; the pots wore covorod with a net and stored In the lab at 23 °C, wotting the soil every two days. Assessments 10 days after the trootment, tho covering nets were removed and the emerged C. varlpos adults were counted RESULTS AND DISCUSSION ■ , S. foltlao application tothelMVOa S. toltlao suspension formulated with an adjuvant, showed some, .Woe ..In controlling C. varlpos Infestation: 76% larval mortality was ach aved. but the dissection of 36 dead larva, a. sample, showed thai only ™J™<£» *■ «f*""''"g tho experiments conditions were rather different from tha natural environment. Tablo 3 - Results of foliar application. Mortality du» A-.'.o-.'juont M>y. 31" Troatmont (U, 1.» i s, r.w« |5xio*| Untmlod oontrol Larval mortality rvllmud I 7H.01Z7 14.01 11.4 to EPN» 57.0 0.0 H. bacteriophora application to tho soil The EPN application to tho soil .1 the most suitable lime. »««*^«» |" coňlrolľng Iho C. var/oes first „en.ra.lon. In the urban oak wood only a adult, -ere collected under Ihe .... In the treated surface, while In the untreated control more than 170 specimen, were counted. Tho laboratory experiment confirmed as ob.arv.d In the park: no edůlt. lerged f.om the treated pot, while 1/ apealn-en. Itaw fron. Ih. untroalad ono. Overlooked important characters in females and infective juveniles of the entomopathogenic family Steinernematidae (Nematoda) Zdeněk Mráček1, Vladimír Půža12 and Alice Hypiová12 1- Institute of 1.1 ,i»niology. Biological Cental. Czech Academy of Science*, Branlsovake 31. 370 OB Cetké Budějovice. Czech Republic I-Faculty or Biologlc.il Science*, Unlver»Ky of South Bohemia, Branltovtka 31, 37006 Cetk* Budějovice, Czech Republic Introduction The family Stememematidae includes more than 40 described species which morphological identification is based on several male and larval character* such as shape of spicule* and guberbaculum in males, position of excretory pore, lateral fields, length of tall and hyaline layer in infecbve juvenile* (Ui) etc Tie" present a lack of tsxawnicalry important characters was found for steinernemaud females (Hormnick et al, 1997). Such analysis of character* * furvtarnental fr/ species distinguish mc. and essential for laxonomk purposes. Therefore, me focused our effort to study steinernernabd female* to ciarry if there are any valuable deferences m the external body morphology (tail protuberances, postanal sweBIng, vulval protruding; among species which could add new important character* to amplify the identification of species and the taxonomy of ths family. Similarly, the 1J tall of different steinernematid species was studied because * belongt at /veil tc tr» tarononvcalh; important characters. Ratio c (tail length/width) and %Hy {length of hyaline layer/tail length x 100) are used in diagnosis and relation*/** of I Both, the methods of scanning electron microscopy and light microscopy were used for our study. Females Tail of the first and second generation Tail shape in the first generation varies among species and partly depends on the age of studied female. In majority of species the ta< rs conical, but blunt in its bp. There b a big difference between females n the first and second yen era Don (Tables 1 and 2). Conical tai tapenng to a pointy end m the second generation females s uniform in most species. ■=3-1—■ .ir.--- T»U» 1. t Jit gnUw f«m*t». rrmprotogy ď lil Vulval region Generally, the first generation possess more protruding vulva than those in the second generation. The vutvel morphology varies among ttewwnemted specie* from slightly to teverafy pvotrutSng (Tsbie 3, Figures) Some speces. such a* S. Yyalnt* width at anus. Till pruaflMrnitoC e*\ morphometry were used in 0V i.im , •«< ■ • However, morpholocKal ta.i diri'.i ••■ appears as more <* lets slender and . >■« attenuated to sutrfigitate atrVxia '-pad** been also wiytooked Postanal swelling Postanal swelling varies from distinct to slight and misting among first generation female specie* (Table 1, Fgures). TTiii character seems to be stable in many ipetie*. E.g. 5 knuul, $■ tthjtKum, S. weaert ha* no or Ught postanal swelling whereat rnott specie* of 5. glattrl group arc ctvaracUsriied by the distinct swelling. The postanal swelling is roduced m the second generation Conclusion: Comparative study of females and Ds tall regions is needed to clarify real significance of above described morphological structures. Especially, female minute tall papilla-like protuberances and general tall character represents a good challenge for future studies. _ Haferanir» II, 1*1 . or LoIaIkvi ami CharBcteMfsHon o! N«y Population* ol Fnlomopiilhogpnlc nomnlodet from I»f»fl1 N MM*.' and I CUturni' C Chktn*Min*hvW' Ave. NOON 'I** ^ j 1,^ q) l«i Thm, rtttH J-«-~ h*w«- a«WM toojton. «■ »» fx— an* .vafuafad «om» taring*! ran h wn> tcmr,. tt«frcoucw- po»»«l O»vcc««on 4 Sajrvti »1»« rip idiri 11' —-' »jt r"*""* 1 111 II ■ - ■ ■ I ' MM ' MM *>.>|»lM*>«'.» (Uaia^laMl) Dm«14U«1 MWIaAM I .IXJKJTIIUSTXWI*- fj r M|M*0 t-, ><^»H> «H I z kiPWtMty; Ew< cox ** Hiwiaan * **> *r *o o TO O eo u so o 40 O ao u 20 O no n o D l ***** Hi I J I Mo****"! w tolumn. MOO Ua «w» Mttl (rtiMlojpl mm Mm iS on gam 11 or ■ CalMWH -i**..Mrl ONI Conclusion! and futura wort. Lanja variation *a* found among tna nt>w nematode population* m r*a#"3 to poflornanc* tucavuy* VM* wtl um INs anaVta •< ban* tot aancton of population for gtmotc improvamant Tna namatodt population, atitrt th» hignaat oatuccaton toUvanca wm b* evaluated agama! foliage psitt 1 Dfimi* ol Meaor* OoM Com* HotpiUI GueoruUrvl Auilnka 2 Otology and Bochemulry. Umverwty ol Balh Balh, UK. The finding that P. asymbiotica exists in an entomopathogenic association similar to other members of the genus has implications for the role of invertebrates in the evolution of emerging human pathogens. The storv so far...... isc I Iff patient has is 'primar • A now P. *symbtottc* Is recovered from a human wound al KlnQSCltff Australia. 2006. • Soil baited al Kingslcift recovers a Heterortwtbdltld MsMMsji • P isymbioucs is recovered from the nematode. 9 The clinical and worm isolates are shown 10 be identical using MLST. • The worm form* a specific symbiosis with both the human and worm derived isolates. (2) A Kinqsctiff worm hunt in EPN complex Is recovered (6) Clinical and worm strains identical In vivo In vitro ~4 Isolation of entomopothogens fro South African soils usin the Galleria 8 Ml |ii>. iimIuii N SVALBARD REINDEER ANTLER CHARACTERISTICS REFLECTING THE LOCAL ENVIRONMENTAL CONDITIONS VERONIKA KAVANOVÁ ' JAN KAVAN 2 (1) Department of Zoology, Facultyof Science, University of South Bohemia (2) Department of Geography, Faculty of Science, Masaryk University i HYPOTHESIS Antler parameters as observed remotely could be used to estimate reindeer population fitness Populations in different localities have different fitness This depends on environmental conditions Locality with advantageous environmental conditions (climate, vegetation) will be inhabited by healthier reindeer population We focused on parameters that could be estimated on distance with use of photography analysis Individuals were photographed from different angles if possible Shoulder height of an individual was estimated (S) and size/length of antler (A) from one single photography -from these parameters the relative size of antler was calculated (L) - see Figure 2 Number of tines was calculated from different photos to ensure the correctness of such number CASE STUDY - Central part of Svalbard (Figure 1) - Comparison of two distinct localities with independent populations - Skansbukta X Petuniabukta - Differences in climate, vegetation and terrain configuration - Reindeers observed during August 2017 Petuniabukt?.: 65 individuals Skansbukta: 94 individuals _ FIG. 1 Relative antler size 40 60 60 100 Cumulative probability % „ „, Skansbukla and Paluniabukla popula.loncbaracletls.lcs based on i Relative antler size I significantly higher in B Skanbukta population 1 Significant shift ■ at around 70% Jj Larger proportion of 3 individuals with I larger antlers In Skansbukta I Missing group of 7-9 linos in I Petuniabukta ^^^^ I IP 12 1 x Skansbukta petuniabukta = 0 7566 m m " 08 0 02 04 0« ľ 0 Relative ant*" «Jflt I .plalive anllw size \v x Skansbukta o Petuniabukta Cumulative probability % mm ^CONCLUSIONS 1 Reindeer population in Skansbukta is in Xbetter Physical condition according to observed j S "e's -relative antler size and number | iodines (Figures 3 and 4) ' This reflects well .better environmental j£J favourable terrain configuration Figure 5) i «-9^*SS^^Bl»^íLh zoology.* Úvod , woakustictó teclinoiogls Modem! bicaiJs" m v komb:naC ibizi noži05' vých projevu ■abyva.ici se ,*, v scuvislo"1 ifj zasaie"6 tou hustotu ,išr těžkým S nízkou intenzitu í výskyt zbytfú lesů ou a pralesoviiy" vozily jscu v oblasti celky s velkým úpením starých CMpnvcn llslnalénebo smläené. s pňlomnostl tekoud p nj mysle n. polního a lei s přirozenou -------- charakterem Z i!eo ska bod v nerha.ino.nějsl menši lesní /asloupenlm starých do.ipnycn s ....._ _.____is^nA c nfl1nmn( |toml iiikuv los n)7ioia»«. íisrsniitl iiPip'WVlI'js piptitrettat 70 'I Tipu tít-.'/uí pv-gmacuj llpdchyt do sltl ['.'y.'''-' ;';j'r: ijitj 1 Nlelod\Ka Na základe následujících Kritérii býlo " oblasti Teslnské pahorkatiny vybráno sest lokalit s předpokladem vysšl koncentrace lesních druhů netopýrů. LeS' listnatý nebo smíšený, bohaté strukturovaný, svySSlm stářím, peslrou skladbou dřevin, přítomnosti všech palet, se etarými a od utni raji dmi Stromy, s potenciami mi netopýřími úkryty a přítomnosti vody siojalé nebo tekoucí. Na vybraných lokalitách byl Instalován stacionární ultrazvukový detektor song meter SM3BAT po dobu ;edné SŽ Sesli noci. Nahrávky by.y zpracovávány pomoci programu SonoChiro. klerý vyuíivá pnn.cp neuronových síti. Terno nasuoi ie ale prozaľ.m potřeba kombinoval S marn-álnl kontrolou nahrávek Jako dop'.nánl metody byl na lokalitách proveden I průzkum heleiodynovým detektorem a na nékterych lokalitách byly provedeny odchyty netopýrů do sir (0ňákOVI*S.7SM79«ll,iaS67S3*lEl Lhravkyz23.-30-6.2019 Eolesicus serotinus 34 i - - r -i:~ -2 12 M i/o tis brandtii 43 Mvotis daubentonii 131 Myotis emorginatus 1 Netopýr velký 1 Myotis tnystacinus 65 Nyctalus leisleri 59 Nyaolus noctolo 96 Pipistrellus nothusjí 8 Pipistrellus pipistrellus 122 Pipistrellus pyqmaeus 17 Pie co t os ouriruí 9 .I Plecatus austriocus 4 1 Vespenilio murinus £- Na Sesli vybráních ta, , Shrnuti ^^^^^^■^■afc-S ÉÄuŕúroby, v. v. i., tým: Funkce HLctlin u aprOSVStČniGCll Clrwsmomum campiiora Nela Glorťková, Helena Ri PROBLÉM Expame iápřednit (QteimMbim . sp.) v íeské Republice / Tyto druhy jsou medializovány / kvůli riiiku jejich kousnutí Ji Zápfcdnice Mildcova osidluje lidská obydli, kde ale není A Značeni potravv pomocí \anthano\du v koloniích čmeláka zemního (Bombus terrestris) a měření konzumace cukru a proteinu při standardních a stresových potravních podmínkách Macháčková L.1'5, Votavové A.2, Mikát Wl.1, Matějková S.\ Řehoř \.36, Gittarová S.4, Straka J.' Katurira zooloaie PřF UK v Praze, Viničná 7, 126 44 Praha contact email; machackovalenkajbclgseznam.cz, Jakub Zemědělský výzkum, spol. s.r.o., Zahradní 1, 66441 Jroubsko-------- - Ústav organické chemie a biochemi 9« - Ustav organické chemie a biochemie AU f-o Piľ„i lrouos,<0. lS5S^Ä=Š£«a?*.--- - Ustav chemického inzenýrsM, Vysoká skola chemicko-lechno/o.i^Le, Technická 3. m 2e, Praná aaculeataresearch.com Složitost nutričního toku a efektivita využívání pylu a cukru v kolonii eusociálních včel nejsou dosud plně pochopeny. Sledovaní pohybu potravy od zdroje až k larvám u včel postupně zásobujících své potomky »■ (progressive provisioning bees) vyžaduje vhodný experimentální přístup. Pomocí lanthanoidových komplexů GdDTPAa DyDTPA (alternativně lanthanoid v komplexu s DOTAs) jsme označili cukr a pyl, jakžto jediné zdroje potravy v kolonii laboratorně chovaného čmeláka zemního (Bombus terres(ris). Porovnali jsme kolonie krmené cukerným roztokem obsahujícím sacharózu s koloniemi krmenými roztokem fruktózy a glukózy (v poměru 1:1). Změřili jsme množství cukru a proteinu zkonzumovaných larvou během vývoje v dospělce a zjistili jsme, jak se mění příjem těchto zdrojů při standardních potravních podmínkách (krmeni ad libitum) a při omezené dostupnosti jednoho ze zdrojů potravy. <3\ □ Metodika spektrometrické techniky ICP-OES 01ICP-M. ^"ektrom'etrické'techniky ICP-OES či ICH-ms. ^ by,á Pro experimenty sestavena.1matky(— em ^ St"ePne ľľÄ'ílternativnej S:,onle byly ^^^5^ monosacharidy ^k^+Ä^. Kolonie s odebíraným py*m n 6 äetk0,onii oplachkokonu i cukru [cl □ Výsledky S Přítomnost lantnanoidů ani typ cukru (muu signifikantní vliv na váhu dělnic a samců. (Obr. 1) Dělnice krmené neomezené přijaty za svůj vývoj v průměru 0.54 mg proteinu a 4.26 mg cukru, samci pak 0.52 mg proteinu a 4.43 mg cukru na 1 mg suché tělesné váhy. Dělnice s omezenou dostupnosti potravy přijaly v průměru 0.51 mg proteinu a 4.65 mg cukru na 1 mg suché tělesné váhy. / Ve všech experimentech byla silná pozitivní korelace mezi váhou jedinců, příjmem * proteinu a prijmem cukru. (Obr. 2) / Konzumace cukru však rostla strměji v porovnáni s konzumací proteinu v koloniích, V kterým byla omezena dodávka potravy (pylu) oproti neomezeně krmeným kolomlm. (Obr. 3) i -k. (rrcharidy , „sacharidy) nemělo """"""" „»»«•'* (jede letny Q Závěr „r, inertního ■ neto»*eh„h, potrat ,,i zapi" ^ ! Jt"os .látek. . .. nsacha^ľ^epotra^P1" , . coMu''"„ov*. lóvaní"1'' cukru Odl krmeny1" *NDOP Downloader - stahování dat z Nálezové databáze ochrany přírody AOPK ČR v prostředí QGIS Kaláb O. Katedra biologie a ekologie PřF OU, Ostrava; OpenGeoLabs s.r.o., Praha Uvod NDOP - Nálezová databáze ochrany přírody AOPK ČR obsahuje záznamy o výskytu druhů od profesionálů i veřejnosti, Tato data jsou v drtivé většině volně dosupné a lze je prohlížet a stahovat pres webový filtr [1], QGIS - Desktopový open-source GIS, který slouží ke správě, prohlížení, analýze a vizualizaci dat [2], NDOP Downloader - Zásuvný modul do QGIS, který nabízí možnost jak pohodlně získat data z nálezové databáze a rovnou s nimi pracovat v prostedí QGIS [3], Pro náročnější uživatele, lze využít samostatný Python modul, nebo nástroj příkazové řádky a tak si práci s databází plné automatizovat. Instalace Instaluje se stadardně jako ostatní zásuvné moduly. Modul je v seznamu veden jako experimentální. Mantis religlosa Spuštění Po instalaci se v horní líště objeví ikonka zásuvného moduluQ. Lze vyhledávat pomocí taxonu (druh, rod) a/ nebo pomocí předdefinovaného regionu, Stažená data Po spuštění se stáhnou a zobrazí všechna dostupná data (vrstvy lokalit i tabulková data k nálezům). Tabulková data se stáhnou ke všem záznamům naraz (narozdíl od webového filtru), u těchto dat se zobrazí souřadnice bodů nebo centroidů, které jsou v tabulce obsaženy Práce s daty SSätmůžeme v 0GIS rovnou > ** d^í^Mobtt na základě Python knihovna a příkazový řádek Pro náročnější uživatele je dostupný Python modul s nástrojem příkazové řádky ndop, který můžeme použít přímo, nebo ho zařadit jako systémový příkaz do workflow v jiném jazyce (např. v R pomocí příkazu systém ()). Python modul umožňuje zadat polygon jako vstupní parametr vymezení území, popř. lze navolit libovolný parametr pro request. Zdroj a manuál Zdrojový kód je volně dostupný na GitHub, kde je i seznam plánovaných funkcí a oprav: https://github.com/OpenGeoI.abs/qg/s-ndop-down/oader Podrobný návod najdete na: https://opengeoiabs.github.io/qg/s-ndop-down/oader/ (1] ftOPK CRiMWt rutotM ouatan ochtvty pr.ajy. hups //porta/./* Deciphering regulatory mechanisms of distinct responses to nutrient Daiances between qeneralist and specialist specialist species ■ Kanaoka' Hirortobu Ucniyar What are I Ol MM .-»ti. liiS *^ttm fttirnjn in nanaiati • I fc! ti KVOTO UNIVF RSI olog of TOFfl/Acttvin I IJ .INJ.II Iff * of Ma i I i«vm ■ ■^pIKalM Oawrife A |D«w| ~ I- 'IM f " Ä ,/ UsvolopinsiiUI (pupation) rula l>> musela Discussion and future plans . r ' ľ i .v 11! I í I (I'llllll'l'i M M itterential Stress Response v? .v m.R-190 |rnR4r*-4M-:jp miR-1003 mfl-313 tfJPJ ......1 hen- Fry- A) OiaMfcation gudeMes of mRNAs uun various denes or audj -m.i__t— ■* 25 Cl-m. stressed (7 days e 33X)and auoar _M (7 oiy. « 2S^c7Ö-l_i^,_X«^ respond cOTerentiy to different stresses /n^-rj7for eornpfe shawd nuctedegenofaaon n 3 etsledcondaon 4. miRNA-137 is expressed in adult Drosophila testes n to the ii*»ype trsto J*" Wave GSCs iGermi™ Stem Cells) we located attached to somatic mehe ce«s called me hub Scale oar iu uni W ^ _ 0_"»a»_Wm,oei «»«_««l.^-".cla».»«W«»ai '"•'S^ 121i^^t51e)™o»aTO and M.m at ■» aiaana. o»t - 'Z^JSSZm it^totn^aKximokaac^oi^U^..(-MaWadll^aiWwM* Tl-Mspenrwaosoons-i J„_Jeo^ed-asocce.* =*«■»>«"0«»™««=« t""" Ihe mauue soeim _ 1 t i in early somatic cyst cells 6. Future Direction & Outlook > Staining for late-stage somatic cells _^^L^_ ' Attempt to rescue testis phenotypes using so Hb^B gal4 driver > Test the permeability barrier S^TS^,,^'**"'"9 tr™'* e*"* " "•* " > Attempt to rescue muscle phenotype Samaine ceil nuvkers on adult n* testes A) Anteoor pan of coni/oi n|i testes shows a lypical level ol somabc cell ««™ii>^iw«mWalonm_,0_1,„ aonormallr lowatdt the huh C) Ouantrf.caiHx, o* «m..T, l^"""" " coo, cons-ol m „*>tl7 mounts When somatic-or germline-si