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1100 години успение на св. Климент Охридски

БИОЛОГИЧЕСКИ ФАКУЛТЕТМЛАДЕЖКА НАУЧНА КОНФЕРЕНЦИЯ

„КЛИМЕНТОВИ ДНИ“17–18 НОЕМВРИ 2016 год.

СБОРНИК С РЕЗЮМЕТА

СУ „Св. Климент Охридски“ – Биологически факултетСъюз на учените в България

Университетски ботанически градини

1100 години успение на св. Климент Охридски

МЛАДЕЖКА НАУЧНА КОНФЕРЕНЦИЯ„КЛИМЕНТОВИ ДНИ“

17–18 ноември 2016 г.София, Биологически факултет

СБОРНИК С РЕЗЮМЕТА

СОФИЯ 2016 г.

НАУЧЕН КОМИТЕТ:Председател:доц. д-р Стоян Шишков - Декан на Биологическия факултет

проф. дбн Росица Конакчиева – СУпроф. дбн Диана Петкова – СУБпроф. д-р Светла Петрова – СУпроф. д-р Христо Гагов – СУ доц. д-р Анели Неделчева – СУ доц. д-р Ганка Чанева – СУдоц. д-р Валентина Ганева – СУдоц. д-р Светослав Димов – СУ доц. д-р Анелия Кенарова – СУдоц. д-р Петя Христова – СУдоц. д-р Албена Гьонова – СУ

ОРГАНИЗАЦИОНЕН КОМИТЕТ:Председател:проф. д-р Мариела Оджакова – СУ

доц. д-р Диляна Николова – СУ доц. д-р Иван Трайков – СУ гл. ас. д-р Анита Тошева – СУгл. ас. д-р Яна Евстатиева – СУгл. ас. д-р Атанас Грозданов – СУгл. ас. д-р Любен Загорчев – СУд-р Красимир Косев - Директор на УБГ;г-н Атанас Згуров - Главен експерт, МОН

Секретар:Мариана Модрева

Графичен дизайн: Димитър Миланов

http://www.biofac.info/

С финановата подкрепа на:Договор № 222/2016 г. Фонд Наука на СУ; Договор № 3022/2015 НИС на СУISSN: 1314–4960

С Ъ Д Ъ Р Ж А Н И Е:

1. Програма .............................................................................................................. 4

2. Пленарни доклади ............................................................................................. 14

3. Постерна сесия ................................................................................................. 19B–Биоразнообразие и биологично образование ......................................... 19M–Молекулярна биология и биотехнологии ................................................ 44E–Екология и устойчиво развитие ............................................................... 86

Младежка научна конференция “клиМентови дни”17-18 ноеМври, 2016 г. , София, БиологичеСки факултет

4

ЧЕТВЪРТЪК, 17 НОЕМВРИ 2016 г.БИОЛОГИЧЕСКИ ФАКУЛТЕТ

АудиторияСЕСИЯЧасФоайе II етажРегистрация08:30 – 12:30

Фоайе I етажМонтиране на постерните табла08:30 – 12:30

Аула239

ОТКРИВАНЕ НА КОНФЕРЕНЦИЯТАПроф. д-р Мариела Оджакова,Председател на Организационния комитетДоц. д-р Стоян Шишков,Декан на Биологическия факултет

1100 години успение на св. Климент ОхридскиСлово на доц. д-р Георги Вълчев, Зам. -Рректор на Софийския университет

09:00 – 9:30

Аула239

ПЛЕНАРНИ ДОКЛАДИЧас Председател – проф. д-р Светла Петрова

Секретар – гл. ас. д-р Любен ЗагорчевProf. Nikolai Zhelev PhD, CBiol, EurProBiol, FIBiol, FMBNAM,FRSBPresident - International BioDiscovery Federation, Director - CMCBR, Abertay University, Dundee, Scotland UKNew Generation Medical Biotechnologies in Drug Discovery and Clinical Practice

09:30 – 10:00

Assoc. prof. Jordan Doumanov PhD, Department of Biochemistry, Sofia University St. Kliment OhridskiCellular Polarity and Retinal Pathologies

10:00 – 10:30

Assoc. prof. Albena Lapeva-Gjonova PhD, Department of Zoology and Anthropology, Sofia University St. Kliment OhridskiSocial parasitism in ants

10:30 – 11:00

Assoc. prof. Ivan Traykov PhD, Department of Ecology and Environmental Protection, Sofia University St. Kliment OhridskiThere and Back Again: A Tale of Gravel Pit Rehabilitation

11:00 – 11:30

Assoc. prof. Asya Asenova PhD, Department of Methodology of Biology Teaching, Sofia University St. Kliment OhridskiCurrent Trends in Professional Training of Pre-Service Biology Teachers

11:30 – 12:00

12.00 – 13.30 ОБЕДНА ПОЧИВКА

ПРОГРАМА


5

Фоайе I етажПОСТЕРНА СЕСИЯ13:30 – 15:30

БИОРАЗНООБРАЗИЕ И БИОЛОГИЧНО ОБРАЗОВАНИЕ

Dimitar Kaynarov, Ivan Andreev, Stela Dipchikova, Boyan ZlatkovDATA ON BUTTERFLIES AND MOTHS FROM THE REGION OF SLAVEYNO, WESTERN RODOPI MOUNTAINS (INSECTA, LEPIDOPTERA)

В1

Milena Nikolova, Ina Aneva, Peter Zhelev, Marina DimitrovaFLAVONOID COMPOUNDS AND ANTIOXIDANT ACTIVITY OF BULGARIAN SPECIES OF MICROMERIA

B2

Venelina Angelkova, Iliyana Pavlova, Ivan Svinyarov, Anita Tosheva, Anely NedelchevaOBSERVATION OF THE WALL FLORA OF UNIVERSITY BOTANIC GARDEN – BALCHIK

B3

Olya AsenovaTHE ROLE OF STUDENT PORTFOLIOS FOR SELF REFLECTION AND IMPROVING STUDENT LEARNING

B4

Cristian BANCIU, Teodora IVANOVA, Gabriel Mihai MARIA, Yulia BOSSEVA, Dessislava DIMITROVA, Anca MANOLECOMPARED MORPHOLOGICAL AND MICROMORPHOLOGICAL CHARACTERISTICS BETWEEN SEEDS FROM SELECTED SCILLA SPECIES AND THEIR TAXONOMIC SIGNIFICANCE

B5

Petya Boycheva, Krasimir KosevETHNOBOTANY OF MEDICINAL PLANTS IN THE REGION NORTH OF VARNA SITY TO DURANCULAC VILLAGE

B6

Dessislava Dimitrova, Teodora Ivanova, Antoaneta Yotova, Rositsa Tropcheva, Emanuela Lukach, Lyudmil HaydutovLOCAL BIODIVERSITY – IMPORTANCE FOR RURAL ECONOMIES AND ADAPTATION TO CLIMATE CHANGE

B7

Nikola Doykin, Elitsa Popova, Neli Ivanova, Diana ZlatanovaACTIVITY PATTERNS AND OCCUPANCY OF THE RED DEER (CERVUS ELAPHUS, L.) AND ROE DEER (CAPREOLUS CAPREOLUS, L.) IN NP VITOSHA, BULGARIA

B8

Violeta Evtimova, Krastio Dimitrov, Venceslav Delov, Yana Velina, Stoyan IordanovDATA ON CETACEAN MORTALITY ALONG BULGARIAN BLACK SEA COAST DURING SPRING-AUTUMN 2016

B9

Tsveta GanevaLEAF EPIDERMIS MORPHOLOGY OF SORBUS UMBELLATA (DESF.) FRITSCH (ROSACEAE)

B10

Galia Georgieva, Yordan Uzunov, Emilia VaradinovaFIRST RECORD OF SIAMESE TWINS IN BRANCHIURA SOWERBYI BEDDARD, 1892 (ANNELIDA: OLIGOCHAETA)

B11


6

Neli Georgieva, Tzvetelina Gergovska, Dimitrina Toncheva, Alexandra Sokolova, Tzvetelina Galabarova, Valya Mashkarova,Lyuba PenchevaMY VOLUNTEER EXPERIENCE AT THE UNIVERSITY BOTANIC GARDENS

B12

Teodora Ivanova, Yulia Bosseva, Chavdar Gussev, Sonya Tsoneva, Valeri Georgiev, Dessislava Dimitrova, Tatyana StoevaCONSERVATION AND RESERCH ON LILIACEA FAMILY IN BULGARIA

B13

Dimitar Kaynarov, Stoyan BeshkovNEW DATA ON RARE AND PROTECTED BUTTERFLIES AND MOTHS FROM BESAPARA HILLS, BULGARIA (INSECTA: LEPIDOPTERA)

B14

Lyubomira Lyubomirova, Plamen Mitov, Lyubomir KenderovWATER MITES (HYDRACHNIDIA) FROM ISKAR RIVER WALLEY (BULGARIA) AND DYNAMICS OF BASIC HYDROCHEMICAL PARAMETERS

B15

Albena Lapeva-Gjonova, Alexander MilushevANTS FROM THE „VRATSA KARST“ RESERVEB16

Mete Mısırlıoğlu, Hristo Valchovski and Ralitsa TsekovaPEREGRINE EARTHWORMS (CLITELLATA: LUMBRICIDAE) FROM BULGARIA AND TURKEY

B17

Silvia Nestorova, Atanas Grozdanov, Miroslav SlavchevMAN-MADE STRUCTURES AS A CONSERVATION TOOL FOR AMPHIBIANS AND REPTILES

B18

Lina Paunova, Dimitar Nenov, Dimitar Dimitrov, Yana Yordanova, Plamen Petrov, Marina Yordanova, Dilyan Minev, Atanas GrozdanovFAUNISTIC OBSERVATIONS IN BOTANICAL GARDENS OF SOFIA UNIVERSITY DURING STUDENT VOLUNTEER ACTIVITIES IN 2016

B19

Plamen Petrov, Atanas Grozdanov, Georgi KrastevPRELIMINARY RESEARCH OF THE HERPETOLOGICAL DIVERSITY OF LYULIN MOUNTAIN, BULGARIA

B20

Petar Petrov, Elitsa Popova, Diana ZlatanovaFOOD PREFERENCES OF THE STONE MARTEN MARTES FOINA, ERX. IN THE MOUNTAINOUS AREA OF TRAN, BULGARIA

B21

Yana PetrovaNEW DATA ON THE DISTRIBUTION OF DORCUS PEYRONIS REICHE & SAULCY, 1856 AND TRYPOCOPRIS AMEDEI (FAIRMAIRE, 1861) (COLEOPTERA:SCARABAEOIDEA) IN BULGARIA

B22

Monica Pramatarova, Ilia GjonovAN ONLINE SCRATCHPADS DATABASE ON THE BULGARIAN JUMPING PLANT-LICE (INSECTA, HEMIPTERA, PSYLLOIDEA)

B23


7

Veselin V. Shivarov, Holger Thüs, Cvetomir M. DenchevECOLOGY AND DISTRIBUTION OF DERMATOCARPON (LICHEN-FORMING FUNGI) IN THE WATERSHEDS OF TWO BULGARIAN RIVERS

B24

Hristo Stoyanov, Valentin BaychevRESEARCH ON THE VARIABILITY IN TRITICALE (XTRITICOSECALE WITTM.) CROSSES AS A SOURCE OF GENETIC DIVERSITY

B25

Bilyana Stoykova, Albena Lapeva-GjonovaCUCKOO BUMBLEBEES AT THE SOFIA UNIVERSITY ZOOLOGICAL COLLECTION

B26

Hristo Valchovski, Mete Mısırlıoğlu and Ralitsa TsekovaFIRST RECORD OF APORRECTODEA JASSYENSIS (MICHAELSEN, 1891) (CLITELLATA, LUMBRICIDAE) FROM RILA MOUNTAIN

B27

Desislava Vladova, Danail Takov, Daniela PilarskaNEW DATA ABOUT PREVALENCE OF MICROSPORIDIUM ENDORETICULATUS POECILIMONAE (MICROSPORIDIA: ENCEPHALITOZOONIDAE) IN THE GRASSHOPPER POECILIMON THORACICUS (ORTHOPTERA: TETTIGONIIDAE)

B28

Stoyan Yordanov, Yana Yordanova, Venceslav Delov, Krustio DimitrovANALYSIS OF THE NEGATIVE IMPACT ON SOME TETRAPODS ALONG BULGARIAN BLACK SEA COAST

B29

Stoyan Yordanov, Stoyan Shishkov, Venceslav Delov, Krastio DimitrovMETHODS TO STUDY THE DISTRIBUTION OF PARAMYXOVIRUSES IN BULGARIAN BATS

B30

Stoyan Yordanov, Venceslav Delov, Krastio DimitrovADAPTIVE STRATEGIES OF SOME BAT SPECIES DURING HIBERNATION IN SOUTH-EAST BULGARIA

B31

Yana Yordanova, Venceslav Delov, Krustio Dimitrov, Stoyan YordanovINVESTIGATIONS ON BIRDS INHABITING THE LITHORAL AREA, TRANSITIONAL WATERS AND ITS SURROUNDINGS OF BULGARIAN BLACK SEA COAST DURING THE BREEDING SEASON

B32

МОЛЕКУЛЯРНА БИОЛОГИЯ И БИОТЕХНОЛОГИИ

Boyka Andonova-Lilova, Tanya Zhivkova, Lora Dyakova, Galina Kurteva, Panaiot Kurtev, Radostina AlexandrovaTO MEET THE CHALLENGES OF PRIMARY CELL CULTURES FROM COLORECTAL CANCER

M1

Nevena Aneva, Gergana Savova, Katia Stankova, Rayna BotevaCUMULATIVE EFFECT OF RADIATION AND INFLAMMATION ON HUMAN BLOOD MONONUCLEAR CELLS

M2

Boyana Angelova, Valentina GanevaELECTROINDUCED RELEASE OF β-GALACTOSIDASE FROM KLUYVEROMYCES YEAST

M3


8

Petia Angelova, Anton Hinkov , Venelin Tsvetkov ,Kalina Shishkova, Daniela Dragolova, Veneta Kapchina-Toteva, Stoyan ShishkovACTIVITY OF WATER EXTRACT FROM NEPETA NUDA L. (LAMIACEAE) AGAINST REPLICATION OF HUMAN HERPES VIRUS TYPE 1 (HERPESVIRIDAE)

M4

Marina Balinska, Iliyan Banev, Martin Dimitrov, Tanya Strateva, Svetoslav G. DimovINVESTIGATION OF SOME OF THE REACTION CONDITIONS ON THE DISCRIMINATORY POWER OF THE RAPD ANALYSES BY UPGMA AND NEIGHBOR-JOINING METHODS USING SOME ENTEROCOCCI AS SUBJECT

M5

Iliyan Banev, Marina Balinska, Martin Dimitrov, Tanya Strateva, Svetoslav G. DimovMOLECULAR CHARACTERIZATION OF SIX ENTEROCOCCUS STRAINS FROM FRESH BEEHIVE POLLEN FROM PIRDOP REGION

M6

Magdalena D. Cetner, Vasilij Goltsev, Katarzyna Kowalczyk, Hazem M. KalajiSPECIFICATION OF NITROGEN AND POTASSIUM DEFICIENCY-INDUCED CHANGES IN PHOTOSYNTHESIS RATE AND ELECTRON TRANSPORT CHAIN OF RADISH

M7

Todor Chaushev, Dimitar Parvanov, Gueorgui Stamenov, Balik DzhambazovGNRH-I – A SUITABLE BIOMARKER FOR ASSESSMENT OF ENDOMETRIAL RECEPTIVITY

M8

Lyubomira Denkova, Maria Dimitrova, Slavil Peykov, Svetoslav G. DimovCONSTRUCTION OF TWO CLASSICAL PLASMID CLONING VECTORS WITH A POSITIVE SELECTION BASED ON THE EXPRESSION OF A BLUE AND YELLOW CHROMOPROTEINS

M9

Maria Dimitrova, Svetoslav G. DimovREVIEW OF SOME ALTERNATIVE CLONING METHODS NOT BASED ON THE USE OF RESTRICTION AND LIGATION ENZYMES

M10

Petya Dimitrova, Gergana Marinova, Svetoslav Alexandrov, Ivan Iliev, Plamen PilarskiBIOCHEMICAL CHARACTERISTICS OF A NEWLY STRAIN COELASTRELLA SP. BGV CULTIVATED AT DIFFERENT TEMPERATURES AND LIGHT INTENSITIES

M11

St. Dimitrova, B. Pavlova, M. Paunov, K. Dankov, V. Velikova, Ts. Tsonev, D. Doneva, M. Kouzmanova, V. GoltsevPROFILING OF PHOTOSYNTHETIC MACHINERY IN LEAVES OF PLANE TREES (PLATANAUS ORIENTALIS) BELONGING TO TWO ECOTYPES DURING HEAT STRESS

M12

Lora Dyakova, Tanya Zhivkova, Georgi Toshev, Daniela-Cristina Culita, Gabriela Marinescu, Luminita Patron, Radostina AlexandrovaA COMBINATION OF DISULFIRAM AND COPPER (II) COMPLEX OF MELOXICAM SIGNIFICANTLY REDUCE VIABILITY AND PROLIFERATION OF HUMAN NON-SMALL CELL LUNG CANCER CELLS

M13


9

Slavil Peykov, Desislava Popova, Nikolay Krumov, Nikolay Gadev, Ilka Tsvetkova, Evgenyia Mihaylova, Natali Atanasova, Gergana Tzankova, Mario Markov, Dimitar Cholakov, Yordanka Kostova, Boris KirovNOVEL TARDIGRADE HEAT-SOLUBLE AND LEA PROTEINS USED AS CRYOPROTECTANTS AND OSMOTIC STRESS PROTECTANTS IN BACTERIA

M14

Rumiana Ganeva , Dimitar Parvanov, Gueorgui Stamenov, Balik Dzhambazov, Todor ChaushevTNF-α EXPRESSION OF IN-VITRO CULTURED HUMAN ENDOMETRIAL STROMAL AND EPITHELIAL CELLS

M15

I. Georgieva, S. Angelova, N. KorsunFLU SEASON 2015/16 IN BULGARIA: ANTIGENIC AND MOLECULAR CHARACTERISTICS OF INFLUENZA VIRUSES

M16

Irina Georgieva, Marina Stantic, Margareta WilhelmSTUDYING THE ROLE OF ∆NP73 IN MITOCHONDRIAL FUNCTION

M17

Teodora Georgieva, Yana Evstatieva, Dilyana Nikolova, Valentin Savov

PLANT GROWTH PROMOTING EFFECTS OF PSEUDOMONAS PUTIDA STRAINS

M18

Vilyana Georgieva, Dimitar Parvanov, Gueorgui Stamenov, Todor ChaushevRELATION BETWEEN MULTIPLE DEFECTS IN HUMAN SPERMATOZOA AND SPONTANEOUS ABORTIONS IN PATIENTS UNDERGOING IVF

M19

Kamelia Hristova-Panusheva, Milena Keremidarska-Markova, George Altankov and Natalia KrastevaDOES AGING AFFECTS AFFINITY OF PRYMERY STEMM CELLS AND OSTEOBLASTIC CELL LINE TO EXTRACELLULAR MATRIX PROTEINS?

M20

Nenko Ivanov, Dragomira Stoyanova, Anita Nikolova, Aleksandra Hristova, Silvia Yaneva, Monika Borisova, Simona Galabova, Todorka Vladkova, Iliana Ivanova, AneliaKostadinovaINTERACTION BETWEEN BACTERIA, KERATINOCYTES CELL LINE AND RGO, AG- RGO COLLAGEN NANOCOMPOSITES

M21

Dimana Kabova, Ralitsa Vassileva, Tania Mitova, Yana Vassileva, Vyara MamatarkovaPRESERVATION OF ACIDITHIOBACILLUS FERROOXIDANS JCM 3863 BY USING A LYOPHILIZED PROTECTIVE AGENT

M22

Petya Koleva, Ina Aneva, Ljuba Evstatieva, Kalina DanovaCOMPARATIVE STUDY OF THE EFFECTS OF PLANT GROWTH REGULATORS AND ACTIVATED CHARCOAL ON THE DEVELOPMENTAL PATTERNS OF SIDERITIS SCARDICA IN VITRO CULTURES

M23

Kristina Lazarova, Vyara MamatarkovaCULTIVATION OF SUSPENDED CELLS CULTURES AND BIOFILM OF ACIDITHIOBACILLUS FERROOXIDANS JCM 3863 IN PRESENCE OF COPPER IONS

M24


10

Emanuela Lukach, Dilyana Nikolova, Yana Evstatieva, Teodora Georgieva, Dessislava Dimitrova, Teodora Ivanova, Rositsa TropchevaANTIBACTERIAL AND ANTIFUNGAL ACTIVITY OF LACTOBACILLUS PLANTARUM STRAINS ISOLATED FROM BULGARIAN GREEN CHEESE

M25

Sylvia Marinova, Tereza vaz Martins, Richard MorrisAN INFORMATION THEORETIC QUANTIFICATION OF CALCIUM SPECIFICITY

M26

K. Mladenova, S. Petrova, T. Andreeva, N. Mladenov, V. Moskova-Doumanova, T. Topouzova-Hristova, Y. Petseva, K. Balashev, Z. Lalchev, J. DoumanovVISUALIZATION OF HBEST-1 PROTEIN BY ATOMIC-FORCE MICROSCOPY

M27

Vella Nikolova, Valya VassilevaRESPONSE OF DNA METHYLATION MUTANTS OF ARABIDOPSIS THALIANA TO HIGH SALINITY

M28

Dimitar Parvanov, Vilyana Georgieva,Gueorgui Stamenov, Todor ChaushevHIGHER FREQUENCY OF TAIL DEFECTS IN HUMAN SPERMATOZOA CORRESPOND TO HIGHER LEVELS OF DNA FRAGMENTATION

M29

M. Paunov, B. Pavlova, E. Chachi, St. Dimitrova, K. Dankov, V. Velikova, Ts. Tsonev, M. Kouzmanova, V. GoltsevCOMPARATIVE INVESTIGATION OF THE MODERATE HEAT STRESS REACTION BETWEEN TWO ECOTYPES OF PLANE (PLATANAUS ORIENTALIS) AND OLIVE (OLEA EUROPAEA) TREES

M30

Tsvetelina Paunova-Krasteva, Vasilka Velkova, Dayana Borisova, Stoyanka StoitsovaINTRACELLULER SURVIVAL OF PSEUDOMONAS AERUGINOSA PAO1 IN A549 CELLS

M31

Yulia Petseva, Kirilka Mladenova, Stoyanka Stoitsova, Jordan Doumanov and Svetla PetrovaEFFECT OF SNAKE VENOM PHOSPHOLIPASE A2 ON ACTIN CYTOSKELETON REARRANGEMENT OF RETINAL PIGMENT EPITHELIUM CELL LINES

M32

Mariya Rogova, Detelin Stefanov, Antoniya Parvanova, Veneta Kapchina-TotevaEX SITU CONSERVATION AND BIOLOGICALY ACTIVE COMPOUDS IN ACHILLEA THRACICA

M33

Izabela A. SAMBORSKA, Vasilij GOLTSEV, Leszek SIECZKO, Hazem M. KALAJITHE COMPARISON BETWEEN MAGNESIUM DEFICIENCY AND SULPHUR DEFICIENCY FOR EARLY DETECTION OF CHLOROPHYLL FLUORESCENCE IN RADISH PLANTS

M34

Gergana Savova, Nevena Aneva, Katya Stankova, Rayna BotevaPROTEASOME INHIBITION CAN MODIFY THE LEVELS OF INFLAMMATORY CYTOKINES IN IRRADIATED BLOOD MONONUCLEAR CELLS

M35


11

Dmitrii Semenovskii, Yana Evstatieva, Dilyana Nikolova, Teodora Georgieva, Valentin SavovPOTENTIAL USE OF PLANT-GROWTH PROMOTING BACTERIA STRAINS FOR SELECTIVE BIOCONTROL

M36

Katerina Serafimova, Sylvia Marinova, Irina Georgieva, Ani Kercheva, Desislava Popova, Miroslava Zhiponova, Ganka Chaneva, Veneta M. Kapchina-TotevaEFFECT OF CYTOKININS AND AUXINS ON GROWTH AND ANTIOXIDANT ACTIVITY IN IN VITRO CULTURED EXACUM AFFINE L.

M37

Slavyana Yaneva Staykova, Rada Staneva, Blaga Rukova, Kristina Nikolova, Draga Toncheva, Gueorgui Stamenov, Savina HadjidekovaPREIMPLANTATION GENETIC DIAGNOSIS FOR MOSAIC FRAGILE CHROMOSOME 16

M38

Denitsa Teofanova, Lyuben Zagorchev, Kristiyana Marinova, Ivanela Albanova, Mariela OdjakovaANTIOXIDANT RESPONSE IN CONDITIONS OF SALT STRESS OF THREE CEREAL CROPS OF THE GENUS TRITICUM

M39

Dragomira Stoyanova, Iliana Ivanova, Todorka VladkovaANTIFUNGAL POTENTIAL OF SOME COLLAGEN-BASED NANOCOMPOSITES AGAINST CANDIDA LUSITANIAE

M40

Joe Davis Velchev, Rossica Marinova, Maria Dimitrova, Galerida Raikova, Svetoslav G. DimovANALYSIS OF THE PHILOGENETIC RELATEDNESS OF BALKAN TROUT SPECIMENS (SALMO TRURRA M. FARIO) FROM DIFFERENT RIVERS ON THE TERRITORY OF SOUTH-WEST BULGARIA

M41

Ralitsa Veleva, Veselina Moskova-Doumanova, Daniela Dragolova, Jordan Doumanov, Tanya Topouzova-HristovaCOMPARATIVE ANALYSIS OF CYTOTOXICITY OF CHLOROFORM EXTRACTS FROM LAMIUM ALBUM L. AND MORPHOLOGICAL ALTERATIONS IN MAMMALIAN CELLS

M42

ЕКОЛОГИЯ И УСТОЙЧИВО РАЗВИТИЕ

Michaela Alexova, Silvena Boteva, Anelia Kenarova, Galina RadevaFUNGICIDE AMENDMENT OF AGRICULTURAL SOILS SELECTS ANTIBIOTIC RESISTANT BACTERIA

E1

Lilia Angelova, Albena Ivanova, Svetlana Momchilova, Albena Momchilova, Liliana MaslenkovaFUNCTIONAL CHARACTERISTICS OF PHOTOSYNTHETIC APPARATUS IN HALOPHYTE PLANT, ERYNGIUM MARITIMUM L.

E2

Asen Asenov, Mariyana Lyubenova AN OLD BEECH FOREST ON KADIEVI MEADOWS, NORTHPIRIN MT – UNIQUE NATURE WEALTH

E3

Radoslava Bekova, Galerida Raikova-PetrovaON THE PATTERN OF CORRELATION BETWEEN THE FECUNDITY, LENGTH, WEIGHT AND AGE OF MUGIL CEPHALUS LINNAEUS FROM THE AREAS OF BULGARIAN BLACK SEA COAST

E4


12

S. Boteva, R. Tzonev, V. Bogoev, K. Dimitrov, Ts. Dimitrova, A. Kenarova, Ts. Komitova, G. Nachev, M. Nikolov, K. Pachedjieva, N. Parleva, D. Simeonovska, V. Stefanov, I. Traykov, R. TsekovaASSESSMENT OF ECOSYSTEM SERVICES OF HEATHLAND AND SHRUB ECOSYSTEMS ON THE TERRITORY OF BULGARIA

E5

Elitza T. Dencheva, Radoy I. IvanovPESTICIDE CONTROL OF ACETYLCHOLINESTERASE ACTIVITY IN SOME INVERTEBRATE ANIMALS AND THE ROLE OF NITRIC OXIDE AS AN ANTIDOTE IN AN ECOTOXICOLOGY ENVIRONMENT

E6

Velmira Dimitrova, Teodora Georgieva, Maria Geneva, Yuliana MarkovskaDETERMINATION OF TOTAL ANTIOXIDANT CAPACITY IN TWO LYCIUM SPECIES GROWN EX VITRO AT HIGH SALINITY IN HYDROPONIC

E7

Konstantin Dobrev, Gabriela Petrova, Daniela Stanoeva, Maya Velichkova, Antoaneta PopovaPHOTOINHIBITION OF PHOTOSYSTEM II RELATED REACTIONS IN THYLAKOID MEMBRANES OF ARABIDOPSIS THALIANA, WT AND MUTANT LUT2, AT ROOM TEMPERATURE

E8

Simona Georgieva, Petya Dimitrova-Mateva, Svetoslav Anev, Nikolina Tzvejkova, Miroslava Zhiponova, Detelina Petrova, Ganka ChanevaCHANGES OF SUPEROXIDE DISMUTASE ACTIVITY IN THE COMMON BEECH AS A RESPONSE TO HERBIVORY

E9

Aygyun Faik, Milena Gerganova, Gabriela Petrova, Maya VelitchkovaCOMPARISON OF PHOTOINHIBITION KINETICS OF THYALKOID MEMBARNES ISOLATED FROM CONTROL AND HIGH TEMPERATURE ACCILMATED TOMATO PLANTS

E10

Milena Gerganova, Aygyun Faik, Eva Sarvari, Maya VelitchkovaPHOTOINHIBITION IN INDUCED DAMAGES OF PHOTOSYNTHETIC APPARATUS OF TOMATO PLANTS (SOLANUM LYCOERSICUM CV M82)

E11

Borislava Gyosheva, Radostina Hristova, Vladimir ValchevHEAVY METAL CONCENTRATION IN SEVERAL WETLANDS SITUATED ON THREE BULGARIAN DANUBE TRIBUTARIES

E12

Rosen Ivanov, Svetlana Bratkova, Anatoliy AngelovANALYSIS OF THE EFFICIENCY OF MICROBIAL FUEL CELLS BASED ON SULFATE-REDUCTION PROCESS, INTEGRATED IN ANAEROBIC WETLANDS

E13

Milena Mitova, Maria Ivanova, Mihail IlievACTINOBACTERIA AS A COMPONENT OF MICROBIAL COMMUNITY FORMED ON ROCK PAINTINGS OF MAGURA CAVE, BULGARIA

E14

Donaltina Vila, Kalina Vila, Dolja Pavlova, Aida BaniANALYSIS OF VIABILITY THROUGH TETRAZOLIUM TEST OF SEEDS OF TWO ALYSSUM SPECIES OF IMPORTANCE FOR PHYTOMINING FROM SERPENTINES IN ALBANIA

E15


13

Ekaterina Pavlova, Emilia Petrova, Yordanka Gluhcheva, Donika Dimova, Nina AtanassovaEVALUATION OF SOME WEIHGT INDICES AND SPERM COUNT AFTER HYPOXIC EXPOSURE OF RATS AND MICE

E16

Simona Peteva, Mariyana Lyubenova, Hristina KalchevaRELATIONSHIP BETWEEN GROWTH INDEX, SPECIFIC LEAF AREA AND LEAF WEIGHT RATIO OF DECIDUOUS ECOSYSTEMS

E17

Francheska Petrova, Veronika Denova, Kalina Pachedjieva, Ivan Traikov, Vladimir Stefanov, Krastio Dimitrov, Ivana IvanovaALLOMETRIC BIOMASS AND CARBON STOCK ESTIMATION IN SHRUB VEGETATION FOR ASSESSMENT OF ECOSYSTEM SERVICES

Е18

Monika Subeva, Lubomir Kenderov,Vesela EvtimovaECOLOGICAL POTENTIAL OF MODEL RESERVOIRS IN BULGARIA

E19

Vesela Yordanova, Albena Ivanova, Albena Momchilova, Svetlana Momchilova, Liliana MaslenkovaCHANGES IN PHOTOSYNTHETIC ACTIVITY AND FATTY ACID COMPOSITION OF PETASITES HYBRIDUS LEAVES UNDER THE INFLUENCE OF ENVIRONMENTAL FACTORS AT VARIOUS PLANT HABITATS

E20

ПЕТЪК, 18 НОЕМВРИ 2016 г.БИОЛОГИЧЕСКИ ФАКУЛТЕТ

Аула239

ПЛЕНАРНИ ДОКЛАДИЧас Председател – проф. д-р Христо Гагов

Секретар – гл. ас. д-р Мартин ДимитроваAinur Kydyrmoldina PhD, Depaprtment of Applied Biology, Kazakh Humanitarian Juridical Innovative University, KazakhstanThe Influence of the Phytopreparation on Cellular Immunity System (EXPERIMENTAL STUDY)

09:00 – 09:30

Makhsat Kudageldin PhD, Depaprtment of Philosophy and Political Science, Kazakh Humanitarian Juridical Innovative University, KazakhstanPhilosophical and Ethical Problems of Biological Research

09:30 – 10:00

Аула239

Кръгла маса "Кариерно развитие"10:00 – 11:30

Водещ: доц. д-р Анели НеделчеваБългаро-американската комисия за образователен обмен„Фулбрайт”

Университетски ботанически градини към СУ „Св. Климент Охридски”ПНО Фонд за дивата флора и фауна

Биовет АД, гр. ПещераФоайе I етажБиоФест11:30 – 14:30

Аула239

ЗАКРИВАНЕ НА КОНФЕРЕНЦИЯТАНаграждаване „Най-добър постер“14:30


14

ПЛЕНАРНИ ДОКЛАДИ

NEW GENERATION MEDICAL BIOTECHNOLOGIES IN DRUG DISCOVERY AND CLINICAL PRACTICE

NIKOLAI ZHELEV

CBiol, EurProBiol, FIBiol, FMBNAM,FRSBPresident - International BioDiscovery Federation, Director - CMCBR, Abertay University, Dundee,

Scotland UK

Modern Medical Biotechnologies such as New Generation Sequencing, Bio-Nanotechnologies, Live-Cell Biosensors and Stem Cell Technologies are revolutionising the process of drug discovery and development and the clinical practice. We have applied the above technologies in rational drug development in oncology and cardiology and the achievements, associated issues and future perspectives in using such technologies will be discussed. We have developed novel nano-biosensors to monitor effectiveness of kinase inhibitors in live cells. Currently cyclin-dependent kinases (CDKs) are considered as targets for drug discovery and development in both cancer and cardiovascular diseases, which are the leading cause of morbidity and mortality worldwide. A number of kinase inhibitors are in clinical development as anti-cancer drugs. The lack of preclinical models physiologically relevant to humans is the main bottleneck in the drug development phase in particular in the cardiovascular area. Recently human embryonic stem cells demonstrated great promise in bringing new and effective cardiovascular treatments to the market through providing an improved testing platform for preclinical drug screening. We have established a novel human “mini heart” assay with embedded nano- sensors, which allows for assessment of the therapeutic potential of novel compounds in cardiovascular diseases such as heart hypertrophy and avoids the contentious use of animal models. The effectiveness of CDK inhibitors as drug candidates for therapeutic intervention in cardiac hypertrophy is currently under investigation.


15

CELLULAR POLARITY AND RETINAL PATHOLOGIES

JORDAN DOUMANOV

Department of Biochemistry, Sofia University St. Kliment Ohridsk

Cellular polarity is a common feature of different cell types: epithelial, neuronal cells and hepatocytes. These cells perform asymmetric distribution of plasma membrane molecules and the polarized organization of organelles and the cytoskeleton. In polarized cells, plasma membrane is divided into apical and basolateral domains separated by tight junctions, having different composition of proteins and lipids and exposed to different environments. In some diseases (incl. retinal pathologies) alteration of the cellular polarity and sorting of specific proteins have been found. Human bestrophin-1 (hBest1) is a member of the bestrophin family of anion channels. It is transmembrane calcium-activated chloride channel, predominantly expressed in the basolateral membrane of retinal pigment epithelium (RPE) cells. Mutations in the protein cause ocular diseases, named Bestrophinopathies, leading to vision lost. Polarity is particularly important for channels that specifically transport ions and other molecules at the basolateral or apical membranes. Hence, the proper basolateral localization of Best1 is important for its function in RPE cells and our data indicate that several disease-causing mutations are associated with altered polarity of hBest1 protein.


SOCIAL PARASITISM IN ANTS

ALBENA LAPEVA-GJONOVA

Department of Zoology and Anthropology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, 8 Dragan Tsankov str., 1164 Sofia , Bulgaria; [email protected]

Social parasitism is the coexistence in the same nest of two social insect species one of which is dependent on the other. It is rather common in social bees, wasps and ants, and therefore may offer unique model systems to study a number of fundamental problems in evolutionary biology. Social parasitic ants exploit, permanently or temporarily, the colonies of other ant species. Many of the species are extremely rare and have restricted distribution ranges although their hosts can be common. Most of them are listed as potentially threatened due to their rarity and dependence on the host. Obligatory host-parasite interactions are commonly classified in four types: xenobiosis, temporary parasitism, dulosis and inquilinism. This review summarizes the knowledge on 40 social parasitic ant species in Bulgaria, their host range and distribution. Two of them are new for the science and four species are new for the Bulgarian fauna.


16


THERE AND BACK AGAIN: A TALE OF GRAVEL PIT REHABILITATION

IVAN TRAYKOV

Department of Ecology and Environmental Protection, Sofia University St. Kliment Ohridski

The majority of lakes found on old maps, have disappeared by melioration, i.e. dewatering of wetlands to gain arable land. In our days most of the rivers in Bulgaria have a more or less constructed and fixed bed. Nevertheless, more man-made lakes are now created than natural ones are disappearing, mostly by the mining of sand and gravel deposits in the floodplains of rivers. Gravel excavation disturbs valuable riparian habitats and after cessation of mining operations, the newly formed lake is left to reclaim naturally. However, gravel pit lakes are inherently poor sites, due to their steep slopes. Alternatively, shorelines can be reclaimed and managed to provide fish and wildlife habitats. The depth, shoreline shape and slope, area, location and number of islands in resulting lakes, and the plant species that revegetate littoral zones can be managed to influence subsequent colonization by animals. Native species of plants generally are preferable to exotics when revegetating disturbed areas. Monitoring of critical habitat features and plant and animal species is necessary to document the effectiveness of reclamation efforts, and to ensure that habitat, once established, is not lost. This presentation describes the rehabilitation of an abundant sand and gravel pit lake, located at the former Negovan marsh site, near Sofia city. The rehabilitation was carried out as a part of the project “Demonstration project for restoration of priority habitat type wetlands and species with European importance along the Lesnovska River (in the vicinity of Negovan village) under the Treaty grant financial assistance №5103020-by-013 from 17.01.2012, funded under the Operational Programme "Environment 2007-2013", priority axis 3 "Biodiversity Conservation”.

CURRENT TRENDS IN PROFESSIONAL TRAINING OF PRE-SERVICE BIOLOGY TEACHERS

ASYA ASENOVA

Department of Methodology of Biology Teaching, Sofia University St. Kliment Ohridski

Keywords: professional development, pre-service teacher education, interactive methods, ICT, serious game Students and academics agree that there is a need to make learning and teaching on pre-service biology teachers more challenging and engaging with integration of ICT in education. This review provides an overview of the current state of research on professional development of pre-service biology teachers. The purpose of this review is to raise awareness of some innovative methods and technology that we apply in professional development of pre-service biology teachers like serious games, drama methods in education and teaching and learning in virtual environment.


17


THE INFLUENCE OF THE PHYTOPREPARATION ON CELLULAR IMMUNITY SYSTEM (EXPERIMENTAL STUDY)

AINUR KYDYRMOLDINA1*, BEKBOLAT ZHETPISBAYEV2, KLARA ZHARYKBASOVA1, KULYASH TAZABAYEVA1, BATIYASH SILYBAEVA1

1–Depaprtment of Applied Biology, Kazakh Humanitarian Juridical Innovative University, Str. Lenina 11, Semey, Kazakhstan 2–Department of Physiological sciences, State Medical University of Semey, Str. Abai 103, Semey, Kazakhstan*Corresponding author: [email protected] Depaprtment of Applied biology, Kazakh Humanitarian Juridical Innovative University, Str. Lenina 11, Semey, Kazakhstan

Keywords: Еminium Regelii extract, a small dose of radiation, remote period, lymphocytes, emotional stress

The article presents the results of research on the effects of an extract of wild medicinal plants eminium Regel to the body of intact and subjected to emotional stress animals on the background of a low dose of gamma-radiation during the long term. The reaction of the animal organism to the effects of radiation, emotional stress and the introduction of the Еminium Regelii extract was evaluated by the total content of leukocytes, absolute and relative content of lymphocytes (including CD3+ CD4+ CD8+), immunoregulatory index (IRI) and the of leukocyte migrationin hibitionre action(LMIR). The total white blood cell count decreased slightly on the irradiated animals, whereas absolute and relative content of lymphocytes were increased. Among T-lymphocytes CD8 + absolute content increased, whereas significantly decreased both absolute and relative content of CD3 + and CD4 +.In the first days after the emotional stress both absolute and relative content of immune cells decreased. The values of the immunoregulatory index and reaction of the response inhibition of leukocyte migration after irradiation decreased slightly. The injection of Еminium Regelii extract in the early stage of the general adaptation syndrome causes a decrease in the total number of lymphocytes, increase the number of CD4+ lymphocytes and leukocytes lymphokine production abilities. Late stage of the general adaptation syndrome is shoed by increased CD3+, CD4+ lymphocytes and immunoregulatory index.

Acknowledgment: This work was supported by grant National Applied Research Project GF4 / 3028 – The development of biotechnological methods of using medicine herbs with the antitumor effect in the production of fermented milk products, Kazakhstan


18


PHILOSOPHICAL AND ETHICAL PROBLEMS OF BIOLOGICAL RESEARCH

M. T. KUDAGELDIN., M. M. TUIEBAYEV

Depaprtment of Philosophy and Political Science, Kazakh Humanitarian Juridical Innovative University, Str. Lenina 11, Semey, Kazakhstan *Corresponding author: [email protected] Depaprtment of Philosophy and Political Science, Kazakh Humanitarian Juridical Innovative University, Str. Lenina 11, Semey, Kazakhstan

Keywords: a biology, a biophilosophy, a bioethics, a philosophy of life

The relevance of the article caused by the fact that biology in the XXI century is among the leaders of modern science. Biotechnologies are increasingly being used in industry and medicine and recently largely determine the way of life of society as a whole. The study of the phenomenon of life is impossible without the use achievements of biology and philosophy. Biology as a science of living nature is designed to contribute to the accumulation of factual information a relatively surrounding world. Many of the problems that are the focus of the study of biology, are complex and contradictory, so it is important to thoroughly thinking through the methodology of research and interpretation. On the other hand, many aspects of which are studied biological sciences are of significant interest for philosophy. This article pursues primarily a search goal: to identify scientific, epistemological and humanistic formation biophilosophy sources, identify its subject and a place in the holistic view of the world, to identify and formulate the basic problems of research.


19

ПОСТЕРИ

B1DATA ON BUTTERFLIES AND MOTHS FROM THE REGION OF SLAVEYNO, WESTERN RODOPI MOUNTAINS (INSECTA, LEPIDOPTERA)

DIMITAR KAYNAROV, IVAN ANDREEV, STELA DIPCHIKOVA, BOYAN ZLATKOV

Department of Zoology and anthropology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: faunistics, Bulgaria, protected species

New faunistical records on butterflies and moths from the region of Slaveyno Village, Western Rodopi Mountains, Bulgaria are given. The data presented are based on material and photographs collected by students from Sofia University during a field practice held at 17–25 July 2016. An important protected species, Parnassius apollo L., was observed.

B2FLAVONOID COMPOUNDS AND ANTIOXIDANT ACTIVITY OF BULGARIAN SPECIES OF MICROMERIA

MILENA NIKOLOVA1, INA ANEVA1*, PETER ZHELEV2, MARINA DIMITROVA1

1Institute of Biodiversity and Ecosystem Research Acad. G. Bonchev 23 Sofia 1113 Bulgaria2University of Forestry, 10 Kliment Ohridsky blvd., 1756 Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: TLC, DPPH, chemotaxonomy

Four Micromeria species naturally distributed in Bulgaria were examined: Micromeria dalmatica Benth, Micromeria frivaldszkyana (Degen) Velen., Micromeria juliana (L.) Rchb. and Micromeria cristata (Hampe) Griseb. Acetone exudates and methanolic extracts of the species were studied for flavonoid compounds by TLC and HPTLC. Sixteen flavonoid compounds were detected by screening in different TLC systems (different sorbents and mobile phases). Flavonoid aglycones – apigenin, luteolin, apigenin 4'-methyl ether, quercetagetin 3,6,7-trimethyl ether were identified in acetone exudates of the examined species. Flavonoid glycosides – kaempferol 3-rutinoside, quercetin 3-rutinoside (rutin), quercetin 3-glucoside/hyperoside, luteolin 7-0-glucoside as well as chlorogenic acid were determined in the methanolic extracts. The methanolic extract of studied species were evaluated also for free radical scavenging activity using DPPH assay. All extracts showed significant antioxidant activity, and these of M. dalmatica and M. frivaldszkyana exhibited the highest activity with IC50 =21.36 µg/mL and IC50 =22.03 µg/mL, respectively.

Acknowledgment: Program for career development of young scientists, BAS Grant № DFNP-67_A1


20

ПОСТЕРИ

B3OBSERVATION OF THE WALL FLORA OF UNIVERSITY BOTANIC GARDEN – BALCHIK

VENELINA ANGELKOVA1*, ILIYANA PAVLOVA2, IVAN SVINYAROV1, ANITA TOSHEVA3, ANELY NEDELCHEVA3

1 – Faculty of Chemistry and Pharmacy, Sofia University 'St. Kliment Ohridski', Sofia, Bulgaria2 – University Botanic Garden – Balchik, Sofia University ‚St. Kliment Ohridski‘, Sofia, Bulgaria3 – Faculty of Biology, Sofia University ‚St. Kliment Ohridski‘, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: North eastern Bulgaria, urban flora, vascular plants, wall flora, wall preservation

Restored and constantly cleaned walls present a unique opportunity for spontaneous colonization by the plants of the diversified surrounding landscape. The aim of this study is to establish species diversity, characteristics and trends about origin and variability of the vascular wall flora in the nine sites of University Botanic Garden – Balchik. During the study are determined more than 80 species and 65 genera and the most common families are Asteraceae (8%), Caryophyllaceae (8%) Poaceae (7%), Fabaceae (4 %), Rosaceae (4 %), Moraceae (4 %) and Amaranthaceae (4 %). Some of the species are accepted as an additional ornamental element and contribute to the complete perception of architectural and historical sites, especially Cymbalaria muralis, Hedera helix, Ficus carica and Polycarpon tetraphyllum. The total representation of alien species on the studied walls is 17% and a few of them are also highly invasive (Ailanthus altissima, Acer negundo, Amaranthus hybridus, Euphorbia maculata, Oxalis corniculata and Oxalis dilenii). The dominant floristic elements are Adv (25%), Kos (16%), Eur-As (16%) and subMed (10%) and the wall flora shows characteristics similar to the Mediterranean wall floras. Acknowledgment: This study was conducted in the Pilot initiative of Balkan Ecological Center – Volunteering in University Botanic Gardens 2016


21

ПОСТЕРИ

B4THE ROLE OF STUDENT PORTFOLIOS FOR SELF REFLECTION AND IMPROVING STUDENT LEARNING

OLYA ASENOVA

Faculty of Preschool and Primary School Education Sofia University “St. Kliment Ohridski”

Keywords: assessment, portfolio, reflection, student learning

Student portfolios are increasingly used for assessing student competences in higher education. The portfolios is a systematic and organized collection of evidence used by the teacher and student to monitor growth of the student’s knowledge, skills, and attitudes in a specific subject area. The portfolios have illustrated the level of insight, maturity, and synthesis of personal and professional qualities that students are capable of achieving. The portfolio may include a wide variety of forms of evidence over time and allows the students to be creative and personalize their learning. In this article explores how portfolio assessment can enable to more authentic assessment that reflects each student's real progress.

B5COMPARED MORPHOLOGICAL AND MICROMORPHOLOGICALCHARACTERISTICS BETWEEN SEEDS FROM SELECTED SCILLA SPECIES AND THEIR TAXONOMIC SIGNIFICANCE

CRISTIAN BANCIU1, TEODORA IVANOVA2, GABRIEL MIHAI MARIA1, YULIA BOSSEVA2, DESSISLAVA DIMITROVA2, ANCA MANOLE1

1 – Department of Plant and Animal Cytobiology, Institute of Biology, Romanian Academy, 296, spl. Independentei, Bucharest, Romania2 – Department of Plant and Fungal Diversity and Resources, Institute of Biodiversity and Ecosystem Research, BAS, 1113 Sofia, 23 Acad. G. Bonchev St., Bulgaria a*Corresponding author: [email protected]

Keywords: Scilla, conservation, scanning microscopy, morphological characteristics

Scilla species are perennial plants, belonging to Liliaceae family, with heights of 15 to 45 cm, flowers in spring or in autumn before leaf growing. Some species are endangered in the national and European Red Lists. For this reason, the species are important for conservation. Some have ornamental use and/or possibly pharmacological properties, given that some species of the genus Scilla present antioxidant properties with beneficial effects on the skin, digestive and circulatory system. Seeds of nine species of Scilla genus have been analyzed trough scanning electron microscopy and selective coloration histology in order to compare the different characteristics of the structures.


22

ПОСТЕРИ

The species studied are: S. scilloides, S. bifolia, S. hyacinthoides, S. litardierei, S. peruviana, S. reverchonii, S. siberica, S. verna and S. autumnalis habituating in very different regions of the globe from Andes Mountains to Danube plain and Siberian forests. Taking into account that seed morphological and micromorphological characteristics are of taxonomic importance this study compares seed peculiarities of nine Scilla species. Further germinative and molecular studies will reveal variability and adaptability qualities of plants.

Acknowledgment: This work was supported by the Romanian Academy project no. RO1567-IBB08/2016 and by Bulgarian Academy of Sciences, Program for career development of young scientists (grant No. 69/2016.

B6ETHNOBOTANY OF MEDICINAL PLANTS IN THE REGION NORTH OF VARNA SITY TO DURANCULAC VILLAGE

PETYA BOYCHEVA1*, KRASIMIR KOSEV2

1* – University Botanic Garden Varna, Sofia University “St. Kliment Ohridski”, Sofia, BG2 – University Botanic Garden Sofia, Sofia University “St. Kliment Ohridski”, Sofia, BG*Corresponding author: e-mail: [email protected]

Keywords: North Black Sea Coast, ethnobotany, medicinal plants, analysis

This study is part of ethnobotanical study of medicinal plants in the North Black Sea Coast, which includes the area from Durankulak village to the Obzor town. Delimiting card was used to floristic regions in Bulgaria. The study was done on residents of the area north of the Varna city near Durankulak village. Field work was conducted in the period April - September 2016. Surveyed are 107 people from 17 settlements. Respondents are people of different sex, ethnicity, age and education. This work presents the results of surveys of small settlements, since the field work is not over. Processed are and summarized the data from the different applications of medicinal plants in the life of the local population. Made literature review of ethnobotanical research in Bulgaria and Europe.


23

B7LOCAL BIODIVERSITY – IMPORTANCE FOR RURAL ECONOMIES AND ADAPTATION TO CLIMATE CHANGE

DESSISLAVA DIMITROVA, TEODORA IVANOVA, ANTOANETA YOTOVA, ROSITSA TROPCHEVA, EMANUELA LUKACH, LYUDMIL HAYDUTOV

1 – Department of Plant and Fungal Diversity and Resources, Institute of Biodiversity and Ecosystem Research, BAS, 1113 Sofia, 23 Acad. G. Bonchev St., Bulgaria*Corresponding author: [email protected]

Keywords: climate change, geographical indications, local plant varieties, rural areas

Extreme climate phenomena that have been witnessed lately in Bulgaria show that climate change is becoming an important issue in our country. Bulgarian state has ratified the Convention on Climate change and Kyoto Protocol, has elaborated the Greenhouse Gas Inventory and the Third Action Plan on Climate Change (2013 – 2020) and has adopted the Law on Mitigation of Climate Change. However Bulgaria is among the very few Member States that does not have a National Strategy for Adaptation to Climate Change. Human-nature relationships in rural areas are among the most vulnerable when climate changes are concerned because the latter directly affect the well being of local communities. In the same time local farming practices and traditional knowledge can become the starting point of bottom-up adaptation strategies. To become successful these strategies need scientific expertise, market support and political recognition. Being directly linked to the local livelihood and reflecting the environmental conditions of a specific territory the food products with protected designation of drigin (PDO) and protected geographical indication (PGI) cast light on the complexity of rural areas and the multidirectional impact of current climate changes. We present here preliminary results from assessment of local biodiversity as a potential for local economies and production of quality agricultural products.

Acknowledgements: This work is supported by ESSEDRA project (EuropaAid 132438/C/ACT/Multi) and the Bulgarian Academy of Sciences under Support for Young Researchers Programme (grant № 69/2016). Field study for the PDO/PGI certification of Bulgarian products is done in cooperation with Momchil Nekov, Member of the European Parliament.

ПОСТЕРИ


24

B8ACTIVITY PATTERNS AND OCCUPANCY OF THE RED DEER (CERVUS ELAPHUS, L.) AND ROE DEER (CAPREOLUS CAPREOLUS, L.) IN NP VITOSHA, BULGARIA

NIKOLA DOYKIN*1,2, ELITSA POPOVA1, NELI IVANOVA1, DIANA ZLATANOVA1

1 - Department of Zoology and Anthropology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria2 – Nature Park Vitosha, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: activity, temporal overlap, occupancy, ungulates

The roe deer and red deer are widely-spread game species with important roles in the ecosystems. Gathering information their activity patterns and habitat occupancy and selection is vital for their management. We investigated these aspects by placing 20 camera traps according to a predefined grid on suitable animal trails in “Vitoshko-Studena” Hunting Enterprise, Vitosha, Bulgaria. The resulting photos were analyzed in terms of activity patterns by fitting kernel density functions to times of activity and estimating the temporal overlap between the two species. Occupancy of both species was analyzed using the program PRESENCE. The results indicate that the roe deer and red deer share very similar activity patterns (90% overlap) and are predominantly crepuscular. The peaks in their activity are typically just after sunrise and a short time before sunset. The occupancy analysis showed that the probability of detection of the red deer is influenced by the proximity of human settlements, and the roe deer’s detection depends on the elevation and the proximity to human trails or roads. This indicates that the two species utilize the available habitats in a different way, but invariably are affected by the anthropogenic factors.

ПОСТЕРИ


25

B9DATA ON CETACEAN MORTALITY ALONG BULGARIAN BLACK SEA COAST DURING SPRING-AUTUMN 2016

VIOLETA EVTIMOVA1, KRASTIO DIMITROV 2*, VENCESLAV DELOV1, YANA VELINA3,

STOYAN IORDANOV1

1 – Department of Zoology and anthropology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria2 – Department of Ecology and EP, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria3 – Ministry of Environment and Water, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: Phocoena phocoena, Tursiops truncatus, Delphinus delphis, Black Sea, Cetacean mortality

The aim of the present study was to gather data on mortality of Harbour porpoise (Phocoena phocoena), Bottlenouse dolphin (Tursiops truncatus), and Common dolphin (Delphinus delphis), the three representatives of Order Cetacea in the Black Sea, using noninvasive approaches. 10 sample areas were monitored, using transects 3 to 8 km long. Sampling have been done from June to October 2016. The transects were passed along two to seven times. 55 full or partial Cetacean standings were registered and analyzed. The prevailing species was the Harbour porpoise (46%). The category T. truncatus/D. delphis was identified in 23% of the cases. The group of all three species was presented by 29% of the found carcasses. In general 37% of the registered individuals belong to the age class up to 1 year. This approximately high proportion of individuals, born in the past year was shown to be due to the input of newborns of the Harbour porpoise. The mortality among Ph. phocoena of age class up to 1 year was significantly higher than those of the age classes above 1 year, as 69% of the found dead Harbour porpoises belong to the age class up to 1 year. Acknowledgment: This work was supported by grant 101/04.2016 of Sofia University „St. Kliment Ochridski“

ПОСТЕРИ


26

B10LEAF EPIDERMIS MORPHOLOGY OF SORBUS UMBELLATA (DESF.) FRITSCH (ROSACEAE)

TSVETA GANEVA

Department of Botany, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: leaf epidermis, stomata, trichomes, Sorbus umbellata

Leaf epidermis morphology of Sorbus umbellata (Desf.) Fritsch was described and a comparison between the two subspecies - ssp. umbellata and ssp. koevessii (Pénzes) Kárpáti was made. Cuticle ornamentation, epidermal cells, stomata and trichomes were observed by light and scanning electron microscopy and analyzed. The length and the width of the epidermal cells and the stomata were measured and the numbers of stomata and trichomes were counted. All data were processed by the statistical variation method. In result differences in the upper epidermis cells outlines, smaller sizes of both upper and lower epidermis cells, higher stomatal frequency and increased trichome frequency on the lower surface were found in the leaves of S. umbellata ssp. koevessii compared to ssp. umbellata. The observed epidermis peculiarities were probably a xerophytic adaptation to the environment conditions.

B11FIRST RECORD OF SIAMESE TWINS IN BRANCHIURA SOWERBYI BEDDARD, 1892 (ANNELIDA: OLIGOCHAETA)

GALIA GEORGIEVA*, YORDAN UZUNOV, EMILIA VARADINOVA

Department of Aquatic Ecosystems, Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences, 2 Gagarin Street, 1113, Sofia, Bulgaria. *Corresponding author: [email protected]

Keywords: Branchiura sowerbyi, Siamese twins, conjoined twins, Bulgaria

Siames twins of the species Branchiura sowerbyi Beddard, 1892 (Oligochaeta, Tubificidae) have been found in a macrozoobenthic sample from Stoykovtzi Reservoir in Bulgaria during the program for monitoring of surface waters in 2012. The conjoined specimens have separate bodies from the head to the V segment and were connected at their body length beginning from the VI segment. The present article reports this first case of such an anomaly in this aquatic oligochaete species.

Acknowledgment: The material was collected within the project “Monitoring of benthic macroinvertebrates in the lakes as part of the national monitoring program of surface waters for 2012”

ПОСТЕРИ


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B12MY VOLUNTEER EXPERIENCE AT THE UNIVERSITY BOTANIC GARDENS

NELI GEORGIEVA1*, TZVETELINA GERGOVSKA1, DIMITRINA TONCHEVA1, ALEXANDRA SOKOLOVA1, TZVETELINA GALABAROVA1, VALYA MASHKAROVA2, LYUBA PENCHEVA3

1 – Faculty of Chemistry and Pharmacy, Sofia University 'St. Kliment Ohridski', Sofia, Bulgaria2 – University Botanic Garden – Balchik, Sofia University ‚St. Kliment Ohridski‘, Sofia, Bulgaria3 – University Botanic Garden – Sofia, Sofia University ‚St. Kliment Ohridski‘, Sofia, Bulgaria*Corresponding author: [email protected], Faculty of Chemistry and Pharmacy, 1 James Bourchier Blvd., Sofia University 'St. Kliment Ohridski', 1164 Sofia, Bulgaria

Keywords: botanic garden, plant collections, volunteering

In the summer of 2016 a cause brings together many followers in the pilot initiative of the Science education council of the Balkan Ecological Center of University Botanic Gardens, Sofia University “St. Kliment Ohridski” - "Volunteering at Botanic Gardens 2016". Volunteering is a challenge and an opportunity for expression. This is the story of students in pharmacy, which combined with the summer holidays satisfaction to help in the work of some of the most attractive and valuable places in our Alma Mater. What is the main reasons to be volunteer? What is the benefit of volunteering? What it is to be part of the team of the Botanic gardens, engaging in a variety of activities and organizing maintenance and promotion of the plant collections? Here, volunteers and their staff supervisors, share their experiences and impressions why volunteering gives a range of useful skills and helps to improve your teamwork and leadership, why the volunteering is the opportunity to be apart of something bigger than yourself and use your civic responsibility for the greater good and why volunteering is unforgettable experience.

Acknowledgment: This study was conducted in the Pilot initiative of Balkan Ecological Center – Volunteering in University Botanic Gardens 2016

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B13CONSERVATION AND RESERCH ON LILIACEA FAMILY IN BULGARIA

TEODORA IVANOVA, YULIA BOSSEVA, CHAVDAR GUSSEV, SONYA TSONEVA, VALERI GEORGIEV, DESSISLAVA DIMITROVA, TATYANA STOEVA

1 – Department of Plant and Fungal Diversity and Resources, Institute of Biodiversity and Ecosystem Research, BAS, 1113 Sofia, 23 Acad. G. Bonchev St., Bulgaria*Corresponding author: [email protected], Institute of Biodiversity and Ecosystem Research, BAS, 1113 Sofia, 23 Acad. G. Bonchev St., Bulgaria

Keywords: seed conservation, bulbous plants, flow cytometry, in vitro cultivation

Liliaceae is one of the most popular plant families with hundreds species used as medicinal, edible and decorative plants. In Bulgarian flora Liliaceae comprises about 140 taxa, most of which are economically important. About half of the Liliaceae taxa in Bulgaria are of conservation importance and 20 are included in the Bulgarian Red Data Book (2011). Current research project aims to explore the characteristics of the important liliaceous plants in Bulgaria and upgrade the information needed for the development of secure ex situ conservation strategies. Seed collections of more than 20 taxa from genera Allium, Fritillaria, Tulipa, Asphodeline, Scilla, Lillium, Asparagus, Ruscus and others were stored for 1-10 years. Seed viability was tested and in vitro cultures were initiated from the produced seedlings. From the tribe Lilieae collections of Tulipa and Fritillaria showed best viability, germinating readily after cold pretreatment at 5° C for one month. Further studies for determination of the genome size, phytochemical profile and other characteristics will be performed.

Acknowledgment: This work was supported by grant from Bulgarian Academy of Sciences, Program for career development of young scientists (grant No. 69/2016).

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B14NEW DATA ON RARE AND PROTECTED BUTTERFLIES AND MOTHS FROM BESAPARA HILLS, BULGARIA (INSECTA: LEPIDOPTERA)

DIMITAR KAYNAROV1, STOYAN BESHKOV2

1Department of Zoology and anthropology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria2National Museum of Natural History, Tsar Osvoboditel Blvd 1, 1000 Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: faunistics, Macrolepidoptera, conservation status

Data on seven species of Macrolepidoptera previously not reported from the area of Besapara Hills, Pazardzhik Region are given. One of them, Dioszeghyana schmidtii, is included in the Annex II of the Habitat Directive of the European Commission. Other six species are known from few localities in Bulgaria. They are: Melitaea ornata, Cleta filacearia, Panchrysia aurea, Behounekia freyeri, Conistra ragusae, Grammodes bifasciata.

B15WATER MITES (HYDRACHNIDIA) FROM ISKAR RIVER WALLEY (BULGARIA) AND DYNAMICS OF BASIC HYDROCHEMICAL PARAMETERS

LYUBOMIRA LYUBOMIROVA1*, PLAMEN MITOV1, LYUBOMIR KENDEROV2,

1 – Department of Zoology and anthropology, Faculty of Biology, Sofia University “St. Kliment Ohridski”,2 – Department of General and applied hydrobiology, Faculty of Biology, Sofia University *Corresponding author: [email protected]

Keywords: water mites, Hydrachnidia, Iskar River, hydrochemical parameters

The present study gives recent data on species composition, distribution and abundance of water mites (Hydrachnidia) from Iskar River valley (Bulgaria). The field trips were carried out during June 2015 and June 2016 on 13 points from Iskar River and some tributaries (Beli Iskar River, Cherni Iskar River and Vladayska River). The following parameters were measured in situ: dissolved oxygen (mg.dm-3), oxygen saturation (%), conductivity (mS.cm-1) and water temperature (°C). Samples were collected from different microhabitats on the river bottom (according ISO 16150), using a hand net for water mites (mesh size 200 µm). The dynamics of hydro-chemical parameters was determined by the altitude (upper part of Iskar River), the organic pollution of the water (in Iskar River downstream of Sofia City) and the self-purification (lower part of Iskar River). Water mites were represented with a total of 39 species (27 species in 2015 and respectively 29 species in 2016). The water mites’ abundance and taxonomic richness were highest in the upper part of Iskar River, where the ecological status of the aquatic ecosystems is highest.Acknowledgments: This study was funded by Project 100/2016 of Sofia University “St. Kliment Ohridski”.

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B16ANTS FROM THE „VRATSA KARST“ RESERVE

ALBENA LAPEVA-GJONOVA, ALEXANDER MILUSHEV


Keywords: faunistics, ants, protected areas

A research on the ants (Hymenoptera, Formicidae) in the „Vratsa karst“ Reserve (Bulgaria, Western Stara planina Mountains) was conducted for the first time in 2016. Pitfall traps, hand searching, sifting and suction sampler were used in four collecting sites (three forests and an open). A total of 39 ant species from 3 subfamilies were recorded, as 22 are new for Vrachanska Mountain. Species composition, zoogeography and conservation value of the ants of „Vratsa karst“ Reserve were discussed.

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B17PEREGRINE EARTHWORMS (CLITELLATA: LUMBRICIDAE) FROM BULGARIA AND TURKEY

METE MISIRLIOĞLU1, HRISTO VALCHOVSKI2* AND RALITSA TSEKOVA3

1. Eskişehir Osmangazi University, Faculty of Science and Letters, Department of Biology, 26480 Eskişehir, Turkey.2. Institute of Soil Science, Agrotechnologies and Plant Protection “N. Poushkarov”, 7 Shosse Bankya Str., 1080 Sofia, Bulgaria.3. Sofia University, Faculty of Biology, Department of Ecology, Dragan Tzankov 8 blvd., 1164 Sofia, Bulgaria*Corresponding author: [email protected]

Key words: Earthworms, Lumbricidae, Peregrine, Bulgaria, Turkey

In this paper we summarize the knowledge on peregrine earthworm fauna from the entire area of Bulgaria and Turkey. The peregrine lumbricids from Bulgaria and Turkey contains 16 taxa belonging to 7 genera. The most common peregrine taxa of Bulgaria are: Aporrectodea rosea (Savigny, 1826), Aporrectodea caliginosa (Savigny, 1826), Aporrectodea trapezoides, Lumbricus rubellus Hoffmeister, 1843, Lumbricus terrestris Linnaeus, 1758 and Octolasion lacteum (Örley, 1881). The most common peregrine earthworm taxa of Turkey are: Aporrectodea caliginosa (Savigny, 1826), Aporrectodea rosea (Savigny, 1826), Dendrobaena veneta veneta (Rosa, 1884), Eiseniella tetraedra (Savigny, 1826), and Lumbricus rubellus Hoffmeister, 1843.


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B18MAN-MADE STRUCTURES AS A CONSERVATION TOOL FOR AMPHIBIANS AND REPTILES

SILVIA NESTOROVA1*,, ATANAS GROZDANOV1, MIROSLAV SLAVCHEV1

1 – Department of Zoology and anthropology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: conservation, amphibians, reptiles

The following work researches the general effect of different types of artificial structures for some species of amphibians and reptiles. The observed cases include both volunteer and project covered conservation actions in several regions of NW and SW Bulgaria. All of the experimental sites were monitored continuously after their construction. Information on species presence and biological and ecological specifics was gathered. Comments of the construction specifics and problems were discussed. As a result of the work, all observed structures were evaluated as successful for providing of favorable conditions for some amphibians and reptiles. The dry-stone walls were constantly inhabited by four species or reptiles (Lacerta viridis, Podarcis erhardii, Dolichophis caspius, Platyceps najadum) and one amphibian (Salamandra salamandra). The artificial ponds and cattle water tanks were inhabited by amphibians (Bufotes viridis, Hyla arborea complex, Rana dalmatina, Pelophylax ridibundus) within the year of their construction (or the first following mating season), which shows their high conservation efficiency. Following these species, predatory reptile (Natrix natrix and Natrix tessellata) was also observed in one of the monitored artificial ponds. In conclusion, the researched structures can be confirmed as highly efficient for many amphibians and reptiles. Therefore they should be included in all possible conservation projects and also promoted as volunteer activities among interested people.

Acknowledgment: Dedicated to the scientist and friend, who inspired our work - Dr. Nikolay Tzankov.

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B19FAUNISTIC OBSERVATIONS IN BOTANICAL GARDENS OF SOFIA UNIVERSITY DURING STUDENT VOLUNTEER ACTIVITIES IN 2016

LINA PAUNOVA1*, DIMITAR NENOV1, DIMITAR DIMITROV1, YANA YORDANOVA1, PLAMEN PETROV1, MARINA YORDANOVA1, DILYAN MINEV1, ATANAS GROZDANOV1

1 – Department of Zoology and anthropology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: conservation education, fauna, flora

Botanical gardens of Sofia University "St. Kliment Ohridski" play an important role of "green islands" for many wildlife species. The great floral biodiversity provides shelter and food resources to a significant number of vertebrates and invertebrates. The present research focuses on some of the species observed in the summer of 2016 during volunteer activities of students, organized by the Balkan Ecological Centre in University Botanical Gardens Balchick and Varna. The diverse faunistic information is related to the individual interests of the involved students and aims mainly to improve their ability to collect information in the field and to analyze it in the form of scientific publications. As a result of the work, 14 species of snails, 2 species of amphibians, 4 species of reptiles and 50 species of birds were identified in the gardens and their vicinity. Some additional information on species habitat preferences and other ecological characteristics was also gathered. The Botanical gardens, which are open for visitors, can be also used to present successfully different animal species and their conservation problems to the public. In the particular case, both of these sites are strategically situated on the Via Pontica migration road, which offers the opportunity for observation of many bird species. The important educational role of the gardens could be seriously improved by enlarging the knowledge of all species which can be found there. The volunteer initiatives of the Balkan Ecological Centre are therefore a useful tool to assist that process.

Acknowledgment: This work is supported by the Balkan Ecological Centre and Faculty of biology, University of Sofia


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B20PRELIMINARY RESEARCH OF THE HERPETOLOGICAL DIVERSITY OF LYULIN MOUNTAIN, BULGARIA

PLAMEN PETROV1*, ATANAS GROZDANOV1, GEORGI KRASTEV1

1 – Department of Zoology and anthropology, Faculty of Biology, Sofia University “St. KlimentOhridski”, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: herpetology, reptiles, amphibians, Bulgaria

Lyulin mountain is part of the Zavalsko-Planska range, connected with Vitosha mountain by the Vladayski col. Although it is situated close to the city of Sofia, there are very few publications concerning the local herpetofauna. The aim of the present work is to determine the actual species richness of amphibians and reptiles in Lyulin mountain, thus laying the ground for future, in-depth studies in this field of science. The research information was mainly gathered in 2015-2016, but additional data from field trips in the last decade was also used. Observations were made by covering random transects, aiming to cover as much territory as possible. All individuals were documented with GPS locations, photographs and details concerning measures, habitats, etc. As a result of the research a total of 6 species of amphibians (Salamandra salamandra, Lissotriton vulgaris, Triturus ivanbureschi, Bombina variegata, Bufo bufo, Rana dalmatina) and 6 species of reptiles (Podarcis muralis, Lacerta agilis, Lacerta viridis, Coronella austriaca, Zamenis longissimus, Natrix natrix) were found. The puddles and small artificial ponds were identified as habitats with the highest species richness. Suitable places for other, still unrecorded species were found along the mountain, which stresses the need for additional studies in the area.

Acknowledgment: This work is dedicated to our friend and colleague Nikolay Tzankov

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B21FOOD PREFERENCES OF THE STONE MARTEN MARTES FOINA, ERX. IN THE MOUNTAINOUS AREA OF TRAN, BULGARIA

PETAR PETROV, ELITSA POPOVA*, DIANA ZLATANOVA

Department of Zoology and Anthropology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: food niche, Martes foina, scat analysis

The stone marten is widely spread in Bulgaria, but its food habits are poorly studied in many parts of the country, especially in the mountainous areas. The aim of this study was to investigate the species’ food preferences based on the analysis of 80 scats opportunistically collected between March and October 2016 in 2 mountainous study areas near Tran, Western Bulgaria (1. Around Tumba peak (1330 m.a.s.l.), north of Tran and 2. Around Vurpela peak (1241 m.a.s.l.), south of Tran). The main food types observed in the scats were: rodents, fruits (pears, cherry plums, blackberries, dog-rose, Cornelian cherry), relatively scarce occurrence of insects (mostly in the southern zone) and others. In two of the scats, traces of scavenging were discovered – claws of carnivorous mammals). The niche breadth was analysed, as well as the similarities with the food preferences of the species in other mountainous parts of the country.

B22NEW DATA ON THE DISTRIBUTION OF DORCUS PEYRONIS REICHE & SAULCY, 1856 AND TRYPOCOPRIS AMEDEI (FAIRMAIRE, 1861) (COLEOPTERA:SCARABAEOIDEA) IN BULGARIA

YANA PETROVA

1 – Department of Zoology and anthropology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author:e-mail: [email protected]

Keywords: Scarabaeoidea, Bulgaria, Lucanidae, Geotrupidae, distribution

During our research of Scarabaeoidea in riverside habitats in SW Bulgaria, several specimens of Dorcus peyronis Reiche & Saulcy, 1856 (Scarabaeoidea: Lucanidae) and Trypocopris amedei (Fairmaire, 1861) (Scarabaeoidea: Geotrupidae) were collected at light near Kalimantsi village, Pirin Mts., and specimens of T. amedei were collected near Parill village, the foothills of Slavyanka Mts. All localities are new records for the investigated species in Bulgaria. Both species have general East-Mediterranean distribution, known at present only from few isolated localities in Bulgaria.

Acknowledgments: This study was supported by SU “St. Kliment Ohridski”, contract № 8/2016.


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B23AN ONLINE SCRATCHPADS DATABASE ON THE BULGARIAN JUMPING PLANT-LICE (INSECTA, HEMIPTERA, PSYLLOIDEA)

MONICA PRAMATAROVA1, ILIA GJONOV2

Sofia University, Faculty of Biology, Department of Zoology and Anthropology*Corresponding authors:email: [email protected], [email protected]

Keywords: Psylloidea, cybertaxonomy, jumping plant-lice, Scratchpads

A Bulgarian psyllids web database is presented. It is created on the basis of Scratchpads platform of the European Distributed Institute of Taxonomy. Taxonomical part of the database consists of 79 jumping plant-lice species from 6 families. All available literature on the Bulgarian psyllids is summarized. Bibliography is included in standardized fields and it is possible to export the references in *.bib or *.ris format. The references are tagged with the names of the concerning species. A special data type is built in order to collect the host-plant data form the literature and from new materials. The database is implemented to add available data for the host-plants. All deposited Psylloidea specimens at the Sofia University Zoological collection are cataloged after DarwinCore standards. Location data of each specimen is visualized on a map. The specimens data is georefered and prepared for publishing in GBIF database. Some live specimen photographs are tagged with species names and locations and are available on the website. New distribution records for several species are provided. All the data is published under Creative Common Attribution license on the website http://www.psylloidea-bg.myspecies.info


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B24ECOLOGY AND DISTRIBUTION OF DERMATOCARPON (LICHEN-FORMING FUNGI) IN THE WATERSHEDS OF TWO BULGARIAN RIVERS

VESELIN V. SHIVAROV1*, HOLGER THÜS2, CVETOMIR M. DENCHEV1

1 – Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences, 2 Gagarin St., 1113, Sofia, Bulgaria2 – Life Sciences Department, The Natural History Museum, Cromwell Road, London SW7 5BD, United Kingdom*Corresponding author: [email protected]

Keywords: Bulgaria, freshwater lichens, Verrucariaceae

A study of genus Dermatocarpon in the watersheds of Demyanitsa River (Pirin Mts) and Malyovitsa River (Rila Mts) was carried out in 2016. The studied area comprises both rivers from the pine belt to the streams and glacial lakes in the alpine belt. The observation of species distribution and measurements of pH, conductivity and surface temperature of the water were made after the snowmelt in June–July, and then repeated again before the snowfall in the beginning of October in twenty-six localities, ranging in altitude from 1091 m to 2464 m. The temperatures of the upper rock surface and near the water were also measured by an infrared thermometer. Additional measurements were made in August only for Malyovitsa River. Four species were found to be restricted to alpine and subalpine environment: Dermatocarpon arnoldianum, D. luridum, D. miniatum var. complicatum, and D. rivulorum. Dermatocarpon arnoldianum is reported for the first time for Bulgaria. Detailed description and comments on its variability are provided. All of the observed members of Dermatocarpon have different ecological requirements. The formation of colonies and the development of thalli in D. rivulorum correlate with the water temperature. The most crowded colonies and largest thalli were found in the coldest measured water (1.7–3.2 °C). The foliose thalli of Dermatocarpon are easier to collect and measure, compared to the other aquatic lichens with crustose thalli, making them a useful tool for monitoring of the environmental changes.

Acknowledgements: This work was supported by “Program for career development of young scientists, BAS”.


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B25RESEARCH ON THE VARIABILITY IN TRITICALE (XTRITICOSECALE WITTM.) CROSSES AS A SOURCE OF GENETIC DIVERSITY

HRISTO STOYANOV1*, VALENTIN BAYCHEV1

1 – Department of Cereals Breeding, Dobrudzha Agricultural Institute – General Toshevo, General Toshevo, Bulgaria*Corresponding author: [email protected],

Keywords: variability, genetic diversity, triticale

Developing genetic diversity is an important element in the breeding process of the cultural plants. It is invariably related to obtaining of new combinations of traits and properties in the plant organism, which is connected to a higher probability for selection of forms with enhanced yield and ecological plasticity. Triticale, being a product of wide hybridization, possesses amphiploid nature which is a reason of its genetic instability. Developing stable lines and varieties is a long process, which implies high breeding pressure in the hybrid progenies. At the same time, the high variation of certain traits is related to the presence of a wider genetic diversity expressed even in the first hybrid generation. In order to determine the possibility of occurrence of higher genetic diversity in hybrid combinations, ten triticale crosses in first hybrid generation were studied. Complete variation analysis by the studied parameters was carried out. Crosses 24/14 (Akord х Bumerang), 28/14 (Akord х Doni 52), 35/14 (Respekt х Irnik) and 36/14 (Respekt х Dobrudzhanets) demonstrated high variation by the greatest number of parameters. Very low variation for a large number of indices was found in crosses 12/14 (Atila х Akord), 13/14 (Atila х Respekt), 26/14 (Akord х Dobrudzhanets) and 34/14 (Respekt х Bumerang). In combination with the mean values of the respective parameters, crosses 28/14 (Akord х Doni 52) and 36/14 (Respekt х Dobrudzhanets) were with highest probability for greater genetic diversity, which makes them valuable breeding material in the improvement work on triticale.


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B26CUCKOO BUMBLEBEES AT THE SOFIA UNIVERSITY ZOOLOGICAL COLLECTION

BILYANA STOYKOVA, ALBENA LAPEVA-GJONOVA


Keywords: faunistics, Bombus, social parasites

Subgenus Psithyrus (genus Bombus) includes cuckoo bee species that are specialized as obligate social parasites in colonies of the true bumble species. They have lost the ability to collect pollen and to rear their brood. As such, these bees do not found their own nests, but instead, invade the host colonies, kills or subdues the founfress and use the resident worker forces. In contrast to the remainders of the genus Bombus, Psithyrus species have no worker caste. Nine cuckoo bee species are known from Bulgaria although collecting data are old and from few locations. In the current study we present new records of Psithyrus species from the Sofia University Zoological Collection.

B27FIRST RECORD OF APORRECTODEA JASSYENSIS (MICHAELSEN, 1891) (CLITELLATA, LUMBRICIDAE) FROM RILA MOUNTAIN

HRISTO VALCHOVSKI1*, METE MISIRLIOĞLU2 AND RALITSA TSEKOVA3

1-Institute of Soil Science, Agrotechnologies and Plant Protection “N. Poushkarov”, 7 Shosse Bankya Str., 1080 Sofia, Bulgaria.2-Eskişehir Osmangazi University, Faculty of Science and Letters, Department of Biology, 26480 Eskişehir, Turkey.3-Sofia University, Faculty of Biology, Department of Ecology, Sofia, Bulgaria*Corresponding author: [email protected],

Key words: Aporrectodea jassyensis, Clitellata, Lumbricidae, earthworms, Rila Mountain.

The paper presents the first record of Aporrectodea jassyensis (Michaelsen, 1891) from Rila Mountain. In the current study, we summarized the knowledge about the distribution of Aporrectodea jassyensis from the Trans-Aegean region – Balkan and Anatolian Peninsula. The species belongs to the Trans-Aegean species group, and probably is semiperegrine, because it is highly adaptive. Aporrectodea jassyensis is native for Southeastern Europe and Asia Minor, beside it is registered, and probably introduced in many countries from the central parts of Europe and Asia. The paper also provided information about the ecology, habitats and zoogeography of Aporrectodea jassyensis.


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B28NEW DATA ABOUT PREVALENCE OF MICROSPORIDIUM ENDORETICULATUS POECILIMONAE (MICROSPORIDIA: ENCEPHALITOZOONIDAE) IN THE GRASSHOPPER POECILIMON THORACICUS (ORTHOPTERA: TETTIGONIIDAE)

DESISLAVA VLADOVA1, DANAIL TAKOV2, DANIELA PILARSKA1,2

1 – New Bulgarian University, Department of Natural Sciences, Sofia, Bulgaria2 - Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: Poecilimon thoracicus, microsporidian infection, Endoreticulatus

New data about infection of Poecilimon thoracicus caused by the microsporidium Endoreticulatus poecilimoni parazitising the gut of the grasshopper are presented. The prevalence of E. poecilimoni was investigated during 2014, 2015 and 2016. It was shown that the infection rates of the entomopathogen increased within the feeding season of the host. The lowest prevalence of E. poecilimoni was recorded in mid June of 2016 (40.0%). The highest infection rate was observed in the beginning of July 2016 (94.4%). No statistical significant differences in the prevalence of E. poecilimoni in males (55.9%) and females (56.4%) of P. thoracicus were detected.


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B29ANALYSIS OF THE NEGATIVE IMPACT ON SOME TETRAPODS ALONG BULGARIAN BLACK SEA COAST

STOYAN YORDANOV, YANA YORDANOVA, VENCESLAV DELOV, KRUSTIO DIMITROV Faculty of biology, University of Sofia, 8 Dragan Tzankov blvd., Sofia, Bulgaria *Corresponding author:E-mail: [email protected]

The studies have been made over the spring and summer of 2016, and some additional data from the previous years has also been used. The observations were made using the line transect method by foot and car. The most affected animals on the roads are the reptiles. The Caspian whipsnake (Coluber caspius), the dice snake (Natrix tessellata), Coronella austriaca, blotched snake (Elaphe sauromates) and the European pond turtle (Emys orbicularis) gave the greatest number of victims. Other observed species were sheltopusik (Ophisaurus apodus), golden jackal (Canis aureus), Eurasian red squirrel (Sciurus vulgaris), red fox (Vulpes vulpes), wildcat (Felis silvestris), long-eared bat (Plecotus sp.), great tit (Parus major), warbler (Acrocephalus sp.) and others. Three fires were registered during the monitoring along the Black sea coast, including one in the Rezovo area, Bolata and one in landfill between Durankulak and Kaliakra. Dead individuals haven’t been found, but in the affected habitats were found spur-thighed tortoise (Testudo graeca), aesculapian snake (Elaphe longissima), sheltopusik (Ophisaurus apodus), balkan wall lizard (Podarcis tauricus), common wall lizard (Podarcis muralis), European ground squirrel (Spermophilus citellus). The disturbance affects mainly the coastal area and the sand beaches on the coastal area. This includes the dunlin (Charadrius dubius), the Kentish plover (Charadrius alexandrinus) and the Eurasian otter (Lutra lutra). The main negative factors affecting the vertebrate fauna are anthropogenic. On first place is the road traffic, second are fires and last comes the disturbance during the summer season.


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B30METHODS TO STUDY THE DISTRIBUTION OF PARAMYXOVIRUSES IN BULGARIAN BATS

STOYAN YORDANOV1, STOYAN SHISHKOV, VENCESLAV DELOV1, KRASTIO DIMITROV1

1 – Department of Zoology and anthropology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, BulgariaCorresponding author: [email protected]

Key words: bats, paramyxoviruses, methodology

Paramyxovirus infections can cause serious diseases in animals and humans. Some of these are zoonotic diseases and are important for human health. Bats are important vectors and reservoirs of such infections. This study was performed in the period 2015-2016. The studies were carried out in the Strandja mountain where in previous studies is registered endemic paramyxovirus strain. Also some areas in other parts of the country have been included. Methods of the study include collecting samples of urine and excrements. The collection of samples was performed using small cotton pads, which are placed in small plastic cuvettes containing buffer or saline, and then immediately have to be frozen. For the purpose of terrain must wear receptacles with dry ice. When collecting samples the researcher have to foloow standard safety rules by using protective equipment. Complying with the rules of purity of genetic samples is also an important moment. Two approaches are used for sampling - invasive and non-invasive. In the invasive approach bats are caught with vertical ornithological nets. As a result of the stress of capture bats almost immediately separated urine and excrements, which should be immediately collected. In order to perform other studies, briefly bats were placed in cloth bags, from which are also collected separately the excrements. In this method the separated urine and feces are in a small amount, which is a problem for the subsequent analyzes. But the method allows exact species identifiation, gender and age group. In noninvasive approach there is two options. In the first case, in refuges of bats directly under their colonies were placed plastic canvases of different sizes – for single bats, small groups and large colonies. They stay 12 hours asylum to cover a full restig period and then the feces were collected. Another non-invasive option that is applied is to sample directly from the guano. This method collects more material compared to invasive method but in many cases not always bats to be defined at the species level. Usually they can be attributed to a specific bat community or bat refuge. Al a result from the later analyzes we proved that fecal samples are significantly more reliable source for viral RNK, whereas in the samples of urine were found only traces.


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B31ADAPTIVE STRATEGIES OF SOME BAT SPECIES DURING HIBERNATION IN SOUTH-EAST BULGARIA

STOYAN YORDANOV1, VENCESLAV DELOV1, KRASTIO DIMITROV1

1 – Department of Zoology and anthropology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, BulgariaCorresponding author: [email protected]

Key words: bats, behavior, hibernation

Studies in this work were carried out during the winter of 2013-2014, 2014-2015, 2015-2016 mainly in the Strandja Mountain and some important winter shelters on the southern coast. The behavior described method Ad libitum as a method to record and analyze the consequences of the behavior and method description and coding of the very behavior when registering motor complexes. Bat colonies are recorded visually by visiting the winter shelters. Bat colonies are registered visually and photographed. In the presence of active bats used equipment and night vision with infrared radiation, to reduce anxiety on bats. Analyzed is the location of the bats, the height from the floor of asylum, the distance from the entrance, are reported abiotic environmental factors in asylum and beyond. In the region of Strandzha mountain are visited the most important hibernakulumi bats. They can be divided into natural habitat, such as caves Great Vapa, Elenina hole Lake, Kerechnitsite, Kale, Cave near the Greec path, etc., and artificial such as tunnels, galleries, buildings and more. In natural habitats are found mostly horseshoe bats. The Greater horseshoe bat and the Lesser horseshoe bat on the one hand and medium-sized horseshoe bats on the other hand show different types of hibernation behavior. As a special hybernaculum identified for Barbastella barbastellus are only tunnels. Myotis myotis was found in the research area as single males that hybernate only in damp walls of abandoned buildings. Myotis capaccinii occupy both types of shelters. Miniopterus schreibersii hibernates only in buildings forming dense colonies on the ceiling of the room. Bechstein's bat was found only in the mine galleries. The majority of natural refuges in Strandja Mountain are occupied mai n ly from horseshoe bats, because of the specific structure of galleries in the m , requiring flight maneuverability. Shelters from atropogenic type are important for bats from famillyVespertilionidae and species diversity of bats at te region in general.


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B32INVESTIGATIONS ON BIRDS INHABITING THE LITHORAL AREA, TRANSITIONAL WATERS AND ITS SURROUNDINGS OF BULGARIAN BLACK SEA COAST DURING THE BREEDING SEASON

YANA YORDANOVA, VENCESLAV DELOV, KRUSTIO DIMITROV, STOYAN YORDANOV

Faculty of biology, University of Sofia, 8 Dragan Tzankov blvd., Sofia, Bulgaria Corresponding author: E-mail: [email protected]

Keywords: Lithoral area, seabirds

The coastal area has very important role in the life cycle of the birds, since they actively use it for breeding, feeding, migration. In the last years human impact on the coast has increased, with many nesting areas being lost because of that. A survey of the coastal birds was completed along the lithoral area of the Bulgarian Black Sea coast. The studies have been made over the spring and summer of 2016, and some additional data from the 2013 – 2016 period has also been used in the study. The observations have been made using the line transect method. The birds have been registered visually, by sound and from photos. We studied the breeding ornithofauna in the lithoral area and its surroundings. This includes the main typical habitats: coastal cliffs, sand and stone beaches, dunes. The surrounding areas includes grasslands, shrublands, forests, agroecosystems and buildings, situated up to 200 m from the coastline. Additionally the registrations of some conservational important bird species in the nearby wetlands are presented. We divided the birds depending their habitats in the following categories: seabirds, which breed primary in the lithoral area; seabirds and waterbirds, which may nest in other areas, but also use the lithoral for breeding; terrestrial birds, which nest in other areas, but may breed in the lithoral; and species of birds, which only use the lithoral for feeding and resting, but do not breed here. Some of the observed species from the first group were the Yelkouan Shearwater (Puffinus yelkouan), found only as migratory and European shag (Phalacrocorax aristotelis), established as nesting in the region of Tiulenovo- Kamen bryag and Kaliakra cape. In the lithoral area from the second group dominant species is the yellow-legged gull (Larus michahellis). Other species like the white wagtail (Motacilla alba), dunlin (Charadrius dubius) and kenthish plover (Charadrius alexandrinus) are also reporded. The third group is most numerous and includes species like Northern Wheatear (Oenanthe oenanthe), European nightjar (Caprimulgus europaeus), European honey buzzard (Pernis apivorus), eastern olivaceous warbler (Iduna pallida), northern raven (Corvus corax) and others. Dominants in the fourth group are the great cormorant (Phalacrocorax carbo) and the black-headed gull (Chroicocephalus ridibundus). The little egret (Egretta garzetta) is common among the Ciconiformes, and at the ports and villages the hooded crow (Corvus corone) is often observed. The studies found that the dunlin is more adaptive than the kenthish plover, dominating over it and displacing it from its breeding habitats.Biggest number of species were observed on deserted beaches without big human impact on them. This shows that the disturbance is important problem for birds, with only small amount of species adapting to it.

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M1TO MEET THE CHALLENGES OF PRIMARY CELL CULTURES FROM COLORECTAL CANCER

BOYKA ANDONOVA-LILOVA1, TANYA ZHIVKOVA1, LORA DYAKOVA2, GALINA KURTEVA3, PANAIOT KURTEV3, RADOSTINA ALEXANDROVA1

1- Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 25, Sofia 1113, Bulgaria2 - Institute of Neurobiology, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 23, Sofia 1113, Bulgaria3 - Specialized Hospital for Active Treatment in Oncology, Sofia, "Plovdivsko Pole" str. 6*Corresponding author: [email protected]

Keywords: colorectal cancer, primary cell cultures, cell cultivation

Colorectal cancer (CRC) is one of primary cell cultures the most common human malignancies. A wide range of experimental models (primary cell cultures, permanent cell lines) are required to study better the biology of CRC as well as to find improved new strategies for its diagnosis, prognosis and treatment. The aim of our study was to get acquainted with the techniques for the establishment of primary cell cultures of human colorectal cancer and to introduce them in our laboratory. Primary cell cultures from six human CRCs were prepared using routine methods of tumor explant cultivation and mechanical digestion followed by trypsin/EDTA treatment. The cells were maintained at 37oC in a humidified CO2 incubator in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and antibiotics. The results obtained by us revealed that: i) Trypsin/EDTA treatment is not a suitable technique for the establishment of CRC cell cultures and should be replaced by another enzyme/enzyme mixture treatment in our further studies; ii) The content of FCS in culture medium has to be minimized in order to suppress the growth of fibroblasts; iii) A cocktail of 4-6 different antibiotics (including penicillin, streptomycin, gentamicin, metronidazole) is required to manage with microbial contamination of tumor samples.

Acknowledgements: This study was funded by the Program “Support of Young Scientists at the Bulgarian Academy of Sciences” - Grant ДФНП-76/27.04.2016.


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M2CUMULATIVE EFFECT OF RADIATION AND INFLAMMATION ON HUMAN BLOOD MONONUCLEAR CELLS

NEVENA ANEVA*, GERGANA SAVOVA, KATIA STANKOVA, RAYNA BOTEVA

Department of Molecular radiobiology, National Centre of Radiobiology and Radiation Protection, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: cytokine, lipopolysaccharide, human peripheral blood mononuclear cells, inflammation, radiation

Ionizing radiation (IR), depending on radiation dose and radiation dose rate, can exert pro- or anti-inflammatory effects in human peripheral blood mononuclear cells (PBMCs), and induce changes in inflammatory and immune responses. In order to analyze radiation-induced changes in inflammatory response, we examined changes in cytokine levels of Interleukin 6 (IL6), Interleukin 8 (IL8), Interleukin 10 (IL10), Tumor Necrosis Factor alpha (TNF α) and Monocyte Chemoattractant Protein-1 (MCP-1) in PBMCs stimulated with Lipopolysaccharide (LPS). LPS acts as a prototypical endotoxin in the immune cells such as lymphocytes, monocytes, dendritic cells, macrophages and B cells, which promotes the secretion of pro- and anti-inflammatory cytokines. Our results showed radiation- and LPS-induced stimulation of production of TNF α immediately after irradiation in contrast to MCP-1 whose levels were shown to decrease 24 hours post irradiation. The production of the rest of the cytokines, namely IL6, IL8 and IL10 was not affected upon radiation exposure in the stimulated by LPS cells.


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M3ELECTROINDUCED RELEASE OF β-GALACTOSIDASE FROM KLUYVEROMYCES YEAST

BOYANA ANGELOVA*, VALENTINA GANEVA

Department of Biophysics and Radiobiology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia*Corresponding author: [email protected]

Keywords: yeast, Kluyveromyces, β-galactosidase, PEF, electroporation

The enzyme β-galactosidase (EC.3.2.1.23), which hydrolyses lactose into its monomers glucose and galactose, has potential applications in food processing industry. Yeast has been considered as an important source of this enzyme from industrial point of view. Pulsed electric field (PEF)-assisted extraction is a relatively new, highly promising method for the recovery of bioproducts from various types of cells. It is based on loss of membrane barrier function under influence of high intensity electric field pulses (electroporation, electropermeabilisation) and subsequent leakage of soluble cytosolic content out of the cells. Recently PEF treatment was successfully applied to recover homologous intracellular enzymes from different yeast species. The aim of this study was to investigate the applicability of PEF treatment applied in a flow mode as an alternative method for extraction of β -galactosidase from Kluyveromyces. A number of strains Kluyveromyces lactis and Kluyveromyces marxianus were tested for their β-galactosidase activity. The experiments on electroinduced extraction were performed with the strains K.lactis 1470 and 2209 having higher specific activity. The cells were subjected to series of pulses of 0.5 – 0.8 ms duration, field intensity in the range of 3 – 4.5 kV/cm at flow rate of 9-27 ml/min. After electrical treatment the cells were diluted in potassium phosphate buffer and incubated for 2-6 h at 37oC. Maximal release of beta-galactosidase (about 90 % from total content) was obtained with strain 1470. A two-step protocol of incubation was developed allowing recovery and partial purification (factor of purification 2.8) of this enzyme. Data obtained in this study let us to suggest that PEF treatment applied in a flow mode can be utilized for recovery of b-galactosidase from Kluyveromyces.

Acknowledgment: This research received funding from the European Union Seventh Framework Programme (FP7/2012-2017) under grant agreement n° 312004.


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M4ACTIVITY OF WATER EXTRACT FROM NEPETA NUDA L. (LAMIACEAE) AGAINST REPLICATION OF HUMAN HERPES VIRUS TYPE 1 (HERPESVIRIDAE)

PETIA ANGELOVA1*, ANTON HINKOV1 , VENELIN TSVETKOV1 ,KALINA SHISHKOVA1, DANIELA DRAGOLOVA2, VENETA KAPCHINA-TOTEVA2, STOYAN SHISHKOV1

1 – Laboratory of Virology, Faculty of Biology, University of Sofia ‘‘St. Kl. Ohridski’’,2- Department of Plant Physiology, Faculty of Biology, University of Sofia ‘‘St. Kl. Ohridski’’, *Corresponding author: [email protected]

Keywords: Human herpes virus type 1, HHV-1, Nepeta nuda L.

Human herpes virus type 1 (HHV-1), a human pathogen, is a member of the large family of Herpesviridae. HHV-1 commonly infects via the lips or the nose between the ages of 6 and 18 months. The viruses are members of the herpesvirus family. HHV-1 has been more traditionally associated with oral-facial infections, although it is now a leading cause of first episode genital herpes and neonatal herpes. It also causes oral ulcers of varying severity from herpes labialis to gingivostomatitis and pharyngitis. Moreover HHV-1 is the leading cause of sporadic encephalitis (HHV-1 encephalitis) and infectious blindness (HHV-1 keratitis). For treating of herpesvirus infections there are about 11 licensed antiherpetic drugs. The most commonly used ones are the nucleoside analog acyclovir, its derivatives and cidofovir. These drugs are usually effective but drug-resistant strains of HHV-1 frequently develop following mutation in either thymidine kinase or DNA polymerase. Moreover, the toxicity associated with some antivirals limits their use. For all these reasons, the search for new antiviral drugs active against HHV-1 which are pointed to other viral targets, and which have less toxicity is of particular interest. For these reasons a special attention is focused on compounds with natural origin. Plant extracts have complex chemical structure, lower cytotoxicity and due to this the occurrence of resistant strains against their action is delayed. The results of studies on natural compounds are specially promising in that several pure substances of plant origin have been shown to exhibit antiviral activity against HHV-1. The genus Nepeta (Lamiaceae), comprises about 250 species distributed in the central and southern parts of Europe, Asia and the Middle East. Nepeta species are widely used in folk medicine because of their antispasmodic, expectorant, diuretic, antiseptic, antitussive, antiasthmatic and febrifuge activities. We are studied the antiviral activity of water extract fom Nepeta nuda L. derived from in vivo propagated plants. The cytotoxicity was tested on Madin Darby Bovine Kidney (MDBK) cell line. Maximal nontoxic concentration (MNC) and cytotoxic concentration (CC50) of the extract was determined by colorimetric method (MTT assays). The results were measured at 48 hour and 72 hour after adding of the extract. Maximal nontoxic concentration (MNC) of the extract determined at 48 hour is ≈ 4 mg/ml and cytotoxic concentration (CC50) is ≈8 mg/ml. Results obtained for MNC and CC50 at 72 hour after adding of the extract are 2 mg/ml and 4.5 mg/ml, respectively. To determine the antiviral activity of the extract against HSV-1 strain F we used modification of MTT assays at low MOI (effect was expressed as % of protection).


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As a long term experiment (results are measured 5-6 days p.i.) we used the values for MNC measured at 72 hour after adding of the extract. Water extract from Nepeta nuda inhibited significantly the replication of HSV-1. The percentage of protection is up to 70 (IC50 is ≈ 0.989 mg/ml). We conduct also a yield-reduction assay at high MOI [10]. As long as this experiment is terminated at the 24th hour this allowed us to use MNC measured at the second day (effect was expressed as % of inhibition). Inhibition yield production reached ≈98 % at 4.5 mg/ml. There was almost no activity at 2 mg/ml. Further we tested the direct inactivating effect of the extract against extracellular form of the HSV-1. The extract did not show any change in the virus titer.

Acknowledgements: This work was financially supported by the grand № 9/2016 of Scientific fund, University of Sofia „St. Kl. Ohridski“, Bulgaria.

M5INVESTIGATION OF SOME OF THE REACTION CONDITIONS ON THE DISCRIMINATORY POWER OF THE RAPD ANALYSES BY UPGMA AND NEIGHBOR-JOINING METHODS USING SOME ENTEROCOCCI AS SUBJECT

MARINA BALINSKA1, ILIYAN BANEV1, MARTIN DIMITROV1, TANYA STRATEVA2, SVETOSLAV G. DIMOV1*

1-Department of Genetics and anthropology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria2- Dept. of Medical Microbiology, Faculty of Medicine, Medical University of Sofia *Corresponding author: [email protected],

Keywords: RAPD-PCR, UPGMA analysis, Neighbor-joining analysis, genus Enterococcus

RAPD-PCR is a strong, versatile and easy to perform method for studying phylogenetic relatedness without the need of expensive and specialized laboratory equipment other than an ordinary thermocycler and agarose gel electrophoresis unit. Despite these advantages the method has some drawbacks, the main of them being the difficult standardization. In this research we try to overcome it using as subjects nineteen Enterococcus strains belonging to different species and one Streptococcus thermophilus strain. Eight E. faecalis strains, five E. faecium strains, two E. hirae strains, two E. durans strains, one E. gallinarum strain, one E. avium strain and one Streptococcus thermophilus strain were subjected to a series of RAPD-PCR analyses aiming to investigate the role of different PCR reaction conditions and factors on the discriminatory power of the method. The impacts on the final results of matrix DNA concentrations, primers concentrations and annealing temperature were studied and it was found that the best discriminatory power was achieved with 0,08 ng/µl matrix DNA concentration, 0,80 µM primer concentration and 40 °C annealing temperatures.


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M6MOLECULAR CHARACTERIZATION OF SIX ENTEROCOCCUS STRAINS FROM FRESH BEEHIVE POLLEN FROM PIRDOP REGION

ILIYAN BANEV1, MARINA BALINSKA1, MARTIN DIMITROV1, TANYA STRATEVA2, SVETOSLAV G. DIMOV1*

1- Department of Genetics and anthropology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria2- Dept. of Medical Microbiology, Faculty of Medicine, Medical University of Sofia*Corresponding author: [email protected],

Keywords: lactic acid bacteria (LAB), genus Enterococcus, beehive pollen

Beehive pollen is known to carry some non-pathogenic lactic acid bacteria, including representatives of the genus Enterococcus. In this study six Enterococcus strains were isolated from beehives from the village Dushantzi, Pirdop region, Bulgaria, a location chosen for the diverse landscapes and rich, mostly natural, ecosystems. With a polyphase approach including classical microbiological methods and molecular approaches it was determined that two of them belong to E. durans, two to E. faecium, one to E. faecalis and another one to E. gallinarum species. The isolates were also checked with molecular methods for the presence of genetic determinants of some common enterococci-related pathogenicity factors and bacteriocins genes. None of the strains gave positive results for the pathogenicity factors while in some of the strains bacteriocins genetic determinants were discovered. These results let us think that these strains are non-pathogenic, that their discovery in the pollen is not due to animal stock or human faecal contamination, and the hypothesis that they are “cultivated” by the bee hives is not to be excluded.


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M7SPECIFICATION OF NITROGEN AND POTASSIUM DEFICIENCY-INDUCED CHANGES IN PHOTOSYNTHESIS RATE AND ELECTRON TRANSPORT CHAIN OF RADISH

MAGDALENA D. CETNER1,*, VASILIJ GOLTSEV2, KATARZYNA KOWALCZYK3, AND HAZEM M. KALAJI1

1–Department of Plant Physiology, Warsaw University of Life Sciences WULS-SGGW, 159 Nowoursynowska Street, 02-776 Warsaw, Poland2 – Department of Biophysics and Radiobiology, Faculty of Biology, St. Kl. Ohridski University of Sofia, 8 Dragan Tzankov Blvd., Sofia, 1164, Bulgaria3 – Department of Vegetable and Medicinal Plants, Warsaw University of Life Sciences WULS-SGGW, 159 Nowoursynowska Street, 02-776 Warsaw, Poland*Corresponding author: [email protected], Department of Plant Physiology, Warsaw University of Life Sciences WULS-SGGW, 159 Nowoursynowska Street, building no. 37, room 0/98A, 02-776 Warsaw, Poland

Keywords: photosynthesis, chlorophyll a fluorescence, OJIP test, electron transport chain, nutrient deficiency

The aim of the research was to study the efficiency of energy transfer in thylakoid membranes of radish plants grown under conditions of nitrogen and potassium deficiencies. Raphanus sativus var. sativus was grown in hydroponics system with modified Hoagland solution under a set of controlled conditions. Nitrogen and potassium deficiency were researched separately. In both experiments, the followings were evaluated: (i) prompt chlorophyll a fluorescence, (ii) net photosynthesis rate (PN); (iii) chlorophyll content, (iv) fresh weight and dry mass accumulation, and (v) chemical compound analysis of leaves. Analysis of selected OJIP test parameters as well as chlorophyll fluorescence induction curve comparison showed that nitrogen and potassium deficiency affect electron transport chain in different ways. Statistically significant changes in some of OJIP test parameters, e.g. a reduction of maximum quantum yield of primary photochemistry (ϕPo) and performance index (PIABS

tot) were observed before the decrease in PN and the appearance of visible symptoms on plants. Results of this research showed that prompt fluorescence measurements might be helpful in diagnosing of nitrogen and potassium deficiency at an early stage stress.


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M8GNRH-I – A SUITABLE BIOMARKER FOR ASSESSMENT OF ENDOMETRIAL RECEPTIVITY

TODOR CHAUSHEV1, DIMITAR PARVANOV1, GUEORGUI STAMENOV1, BALIK DZHAMBAZOV2

1 – Nadezhda,Women’s Health Hospital, 3 “Blaga vest” Street, Sofia, Bulgaria2 – Department of Developmental Biology, Plovdiv University “Paisii Hilendarski”, Plovdiv, Bulgaria*Corresponding author: Todor Chaushev, [email protected]

Keywords: GnRH-I, human endometrium, endometrial receptivity, implantation window

The hypothalamus and pituitary are the main origin and target sites of gonadotropin releasing hormone (GnRH), but numerous studies have demonstrated that GnRH-I and GnRH-II exist in different reproductive tissues such as the ovary, endometrium e.t.c. The major effects of GnRH-I in the reproductive tissues are the regulation of cell proliferation, activation of apoptosis and remodulation during the process of embryo implantation.

In this retrospective study endometrial samples were obtained from 30 women with normal menstrual cycles. Patients with endometriosis, pelvic inflammatory disease, polycystic ovarian syndrome, age > 40, BMI > 30 and bad quality embryos were excluded from the study. Patients were divided into two groups: (1) women with successful implantation (n=15) and (2) women with unsuccessful implantation (n=15). The stage of menstrual cycle was established by the previous menstrual period, transvaginal ultrasonographic finding and LH surge. Histological and immunnohistochemical dating was used to verify the menstrual dating. Using immunohistochemistry, we identified GnRH-I in stromal, luminal and glandular epithelial cells. The level of protein accumulation (H) was scored as 0 (no staining), 1 (weak staining), 2 (moderate staining) and 3 (strong staining). Student T-test was performed using SPSS version 21.0 (SPSS Inc., Chicago, IL, USA) in order to compare the mean score of GnRH-I. P < 0.05 was considered as statistically significant. Results were reported as mean values.

Our results show that GnRH-I is expressed in different cell types of the human endometrium during the mid-luteal phase (window of implantation) – luminal epithelial cells, glandular epithelial cells and stromal cells. The reaction intensity was significantly different between the studied patient groups. Significantly higher expression of this hormone in the patients with successful implantation was detected in the gland (Group 1 - 2.07 vs. Group 2 - 1.75., P = 0.042) and luminal epithelial cells (Group 1 - 2.05 vs. Group 2 - 1.73, P = 0.049). While оnly 30% of the patients with unsuccessful implantation had values for glandular epithelial cells H ≥ 2, in the patients with successful outcome more than 70% exceeded this value.

As a conclusion an insufficient expression of GnRH-I in human endometrium during the mid-luteal phase could be related to embryo implantation failure. The immunohistochemical assessment of GnRH-I expression should be considered as a suitable biomarker for endometrial receptivity during the window of implantation.

Acknowledgment: This work was supported by Nadezhda,Women’s Health Hospital, Sofia, Bulgaria


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M9CONSTRUCTION OF TWO CLASSICAL PLASMID CLONING VECTORS WITH A POSITIVE SELECTION BASED ON THE EXPRESSION OF A BLUE AND YELLOW CHROMOPROTEINS

LYUBOMIRA DENKOVA, MARIA DIMITROVA, SLAVIL PEYKOV, SVETOSLAV G. DIMOV *

Department of Genetics, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected],

Keywords: plasmid cloning vectors, chromoproteins

Two novel plasmid cloning vectors, the pBlue and the pYellow, were constructed on the basis of the pUC19 cloning vector. They are very similar to their ancestor from which they were derived concerning the mode of recombinants selection. However, due to the replacement of the lacZ’ gene by fusion PCR with the amilCP and amilGFP coding sequences obtained from the coral Acropora millepora, which were codons-optimized for expression in Escherichia coli, the need of X-gal for colour reaction substrate is eliminated and the non-recombinant transformants change the colour of the colonies to blue or yellow respectively. The new vectors showed similar transformation efficiencies as the pUC19 vector, nonetheless the appearance of the colouration took more time in comparison to the classical blue-white screening.

Acknowledgment: This work was supported by Sofia University “St. Kliment Ohridski” Research Fund Grant №. 69/2016


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M10REVIEW OF SOME ALTERNATIVE CLONING METHODS NOT BASED ON THE USE OF RESTRICTION AND LIGATION ENZYMES

MARIA DIMITROVA, SVETOSLAV G. DIMOV*Department of Genetics, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: ligase- and restrictase-independent cloning methods

In this review we examine some new modern cloning methods which circumvent the classical methods based on the use of restriction and ligation enzymes (conventional “cut and paste DNA” cloning) which present many drawbacks despite their historical significance. All of the methods described here require PCR amplification of the target fragment and/or the cloning vector and depending on the part of the original method it eliminates, they can be sorted into three categories – ligation-independent based, restriction-free based and enzyme free. These three groups are fluid and the methods in each group can easily be modified to fit into any of the other two remaining classes. This interchangeability lets a researcher switch between methods mid-experiment if the need arises. These methods are relatively cheap and are they less labour intensive, which makes them a great alternative to the conventional cloning and commercially available cloning kits.

Acknowledgment: This work was supported by Sofia University “St. Kliment Ohridski” Research Fund Grant №. 69/2016


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M11BIOCHEMICAL CHARACTERISTICS OF A NEWLY STRAIN COELASTRELLA SP. BGV CULTIVATED AT DIFFERENT TEMPERATURES AND LIGHT INTENSITIES

PETYA DIMITROVA1*, GERGANA MARINOVA1, SVETOSLAV ALEXANDROV1, IVAN ILIEV1, PLAMEN PILARSKI1

1 - Institute of Plant Physiology and Genetics – Bulgarian Academy of Sciences.Acad. G. Bonchev str. Bl.21, Sofia, Bulgaria*Corresponding author: [email protected]

Key words: Coelastrella, cultivation, temperature, light intensity, metabolites

The adaptive capacity of microalgae to varying temperatures is of key importance for their mass outdoor cultivation because daily and seasonal temperature fluctuations are significant. Our aim was studying the influence of temperature and light intensity on the growth and cellular metabolites of a newly isolated Coelastrella sp. strain BGV. The alga was cultivated at wide temperature range (20 - 44ºC) and two light intensities (unilateral - 8000 lx and bilateral illumination – 2 x 8000 lx). Optimal temperature for algal growth was 25ºC and 35ºC at low and high light intensity, respectively. Regardless of the illumination, the extreme temperatures (40, 42 and 44ºC) were stress factor and led to algal growth retardation. Coelastrella sp. BGV required lower light intensity for biomass accumulation (7,7 g.l-1 dry weight at 25ºC) and temperatures above 35ºC for enhanced protein synthesis. Carbohydrates reached their maximum at 30ºC - 50,7% and 45,7% of DW. Lipids were the most affected by the temperature and light intensity metabolites. They varied from 7,3% to 40% of DW at unilateral illumination and from 10,3% to 37% for bilateral illumination. According to gas-chromatographic analysis the main fatty acids were myristic (14:0), palmitic (16:0), stearic (18:0), oleic (18:1), linoleic (18:2), alpha-linolenic (α-18:3) and their proportion clearly varied according to different growth conditions. Carotenoid production increased with the raising of temperature. Coelastrella sp. BGV showed high productivity and preserved good qualitative composition of the biomass in the range 20-35 оС. The strain could be recommended as a promosing for intensive mass outdoor cultivation.

Acknowledgements: This work was financially assured by "Program for career development of young scientists, BAS, 2016".


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M12PROFILING OF PHOTOSYNTHETIC MACHINERY IN LEAVES OF PLANE TREES (PLATANAUS ORIENTALIS) BELONGING TO TWO ECOTYPES DURING HEAT STRESS

ST. DIMITROVA1, B. PAVLOVA1, M. PAUNOV1, K. DANKOV1, V. VELIKOVA2, TS. TSONEV2, D. DONEVA2, M. KOUZMANOVA1, V. GOLTSEV1

1 - Department of Biophysics and Radiology, Faculty of Biology, St. Kl. Ohridski University of Sofia, 8 Dragan Tzankov Blvd., Sofia, 1164, Bulgaria2 - Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Acad. G. Bontchev Str., bl. 21,1113 Sofia, Bulgaria

Plane (Platanus orientalis) is widely used as a park tree in Europe. While it improves the city microclimate it is considered almost extinct in the natural ecosystems, because of climate changes that cause environmental stress such as abnormally high temperatures, drought and excessive light. Thus it is important to study the plant’s tolerance and adaptive potential at different conditions. The best method for establishing the plants response to environmental stress is measuring of the fluorescence, emitted by chlorophyll a molecules. This method gives fast and very detailed information about the state of the photosynthetic machinery, which is a highly sensitive indicator of the plant physiological state. The analyzed effects depend on the age of the investigated leaves thus in order to correctly interpret the results it is important to analyze leaves from different position from the apical bud and assess the difference in the fluorescent signals, measured at each leaf position. In the present study we have analyzed the effects of moderately elevated temperatures on the photosynthetic characteristics of two ecotypes of Platanus orientalis with Bulgarian and Italian origin, and we have compared how these changes are expressed in leaves of different number from apical part of the investigated trees. The Bulgarian ecotype shows higher tolerance and is less affected by the elevated temperatures in contrast to the Italian ecotype that was more sensitive. Younger leaves of the Bulgarian ecotype bear the temperature changes better than older leaves that are not damaged, but have lower photosynthetic productivity. The Italian ecotype is more affected by the moderate heat stress, but the difference between younger and older samples is less pronounced.

Acknowledgment: We are grateful to Bulgarian National Scientific Fund (Project № DFNI B02/8) for financial support.


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M13A COMBINATION OF DISULFIRAM AND COPPER (II) COMPLEX OF MELOXICAM SIGNIFICANTLY REDUCE VIABILITY AND PROLIFERATION OF HUMAN NON-SMALL CELL LUNG CANCER CELLS

LORA DYAKOVA1, TANYA ZHIVKOVA2, GEORGI TOSHEV2,3, DANIELA-CRISTINA CULITA4, GABRIELA MARINESCU4, LUMINITA PATRON4, RADOSTINA ALEXANDROVA2

1 - Institute of Neurobiology, Bulgarian Academy of Sciences, Acad. Georgi Bonchev Str., Block 23, Sofia 1113, Bulgaria2 - Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences, Acad. Georgi Bonchev Str., Block 25, Sofia 1113, Bulgaria3 - Faculty of Medicine, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria4 - Institute of Physical Chemistry “Ilie Murgulescu”, Romanian Academy, Bucharest, RomaniaCorresponding autor: [email protected],

Keywords: non-small cell lung cancer, meloxicam, disulfiram, cytotoxic/cytostatic effect

Lung cancer is the leading cause of cancer deaths in women and men throughout the world. Non–small cell lung cancer accounts for ~85% of all lung cancer cases. The selective non-steroidal anti-inflammatory drug meloxicam and Cu (II) complex were found in our previous investigations to inhibit the in vitro growth of cultured cell lines established from human (cervical carcinoma and glioblastoma multiforme), rat (sarcoma) and chicken (hepatoma) cancers. Disulfiram, used for decades in the treatment of chronic alcoholism, was proved to express also promising antineoplastic properties. The aim of the present study was to evaluate the influence of disulfiram, Cu (II) complex of meloxicam and their combinations on viability and proliferation of human non-small cell lung cancer cells. The permanent cell line A549 reported to express the cyclooxigenase 2, was used as a model system. Disulfiram was applied at concentrations of 03-100 µg/ml; meloxicam and its Cu (II) complex were administered at a concentration range of 10-500 µg/ml. The investigations were performed using methods with different molecular/cellular targets and mechanisms of action such as MTT test, neutral red uptake cytotoxicity assay and double staining with acridine orange and propidium iodide. The experimental data obtained revealed that a combination of Cu(II) complex of Meloxicam with Disulfiram (applied at relatively low toxic concentrations of 6.25 and 12.5 µg/ml) results in significantly higher cytotoxic/cytostatic effects as compared to the single application of these compounds.

Acknowledgements: This study was funded by the Program “Support of Young Scientists at the Bulgarian Academy of Sciences” - Grant ДФНП-74 А-1 and a bilateral project between Bulgarian Academy of Sciences and Romanian Academy.


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M14NOVEL TARDIGRADE HEAT-SOLUBLE AND LEA PROTEINS USED AS CRYOPROTECTANTS AND OSMOTIC STRESS PROTECTANTS IN BACTERIA

SLAVIL PEYKOV1, DESISLAVA POPOVA1, NIKOLAY KRUMOV1, NIKOLAY GADEV1, ILKA TSVETKOVA1, EVGENYIA MIHAYLOVA1, NATALI ATANASOVA1, GERGANA TZANKOVA1, MARIO MARKOV2, DIMITAR CHOLAKOV2, YORDANKA KOSTOVA3 ; BORIS KIROV2

1- So fia university "St. Kliment Ohridski"; 2- Tehnical University;3-Plovdiv University "Paisii Hilendarski"*Corresponding author: [email protected] - Николай Гъдев, iGEM Bulgaria

Keywords: lyophilization, cryoprotective potential, desiccation resistance, osmotic stress, bacteria, survival rate

Tardigrades are a group of microscopic animals that are well known with their abilities to withstand extreme conditions including freezing, dehydration and osmotic stress. Proteins that were identified to be responsible for these properties include CAHS (Cytoplasmic Abundant Heat Soluble), SAHS (Secretory Abundant Heat Soluble), MAHS (Mitochondrial Abundant Heat Soluble) and RvLEAM (Late Embryogenesis Abundant Protein). Such proteins are usually characterized by an intrinsically disordered structure. All of the listed were found to be tardigrade-unique. As a mechanism It was proposed that they prevent protein aggregation inside the cells when put in a state of dehydration. Expressing them in a prokaryotic cell like the model E. coli may increase its survivability when subjected under extreme conditions as freezing and drying. This may benefit processes like lyophilization in which the productivity is lowered due to the low survival rate of the cells. Our project is an attempt to mimic the tardigrades unique survivability in different bacteria species via heterologous expression of protective proteins. We took the four previously mentioned proteins, selected their longest isoforms and optimized these sequences for expression in E.coli. Those synthetic genes were consequently used for the construction of expression cassettes, which were controlled by a tac promotor (repressible by lacI) and inserted in pSB1A3 vectors. We took the generated constructs and electroporated them into Rosetta gammi strain (optimized for protein expression). Expression levels were monitored with SDS-PAGE. The cryoprotective potential of our devise was tested in freeze and thaw experiments followed by cell viability quantification.


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M15TNF-α EXPRESSION OF IN-VITRO CULTURED HUMAN ENDOMETRIAL STROMAL AND EPITHELIAL CELLS

RUMIANA GANEVA1 , DIMITAR PARVANOV1, GUEORGUI STAMENOV1, BALIK DZHAMBAZOV2, TODOR CHAUSHEV1

1 – Nadezhda,Women’s Health Hospital, 3 “Blaga vest” Street, Sofia, Bulgaria2 – Department of Developmental Biology, Plovdiv University “Paisii Hilendarski”, Plovdiv, Bulgaria*Corresponding author: Rumiana Ganeva, [email protected]

Keywords: TNF-alfa, 3D culture, human endometrial cells, implantation window

The human endometrium is a complex tissue composed of various cell components, which interaction determine its main functions in relation to embryo adhesion and implantation. During each menstrual cycle, the human endometrium undergoes a series of well controlled changes in anticipation of the arrival of the blastocyst that are still not completely understood. The actual role of TNF- α in endometrial function is not yet completely understood. Previous analyses have shown the presence of TNF-α and its receptors within both the stromal and epithelial cells of the human endometrium. TNF-alpha has a specific menstrual cycle-dependant expression which is highest during the secretory phase. This cytokine has a local role in a variety of endometrial functions in order to support embryo implantation, including cell differentiation and tissue remodelling.

In the present study, we examined the secretion of TNF-α by primary human endometrial stromal (ESC) and epithelial (EEC) cells cultured in RPMI1640 media, supplemented with 10% FBS. Endometrial epithelial and stromal cells were obtained from mid-secretory stage endometrial samples from 28 healthy women and cultured in vitro in 3D Matrigel (BD Biosciences) for 48 hours. The presence of TNF-α in the culture media was measured using enzyme-linked immunosorbent assay (ELISA). Absorbance was read at 450 nm using a DTX 880 Multimode Detector (Beckman-Coulter).

Cell culture measurements revealed that epithelial cells secrete higher quantities of TNF-α (2.2 pg/ml ± 0.7 pg/ml), compared to stromal cells (0.5 pg/ml ± 0.2 pg/ml). Moreover, co-culture of epithelial and stromal cells showed the highest expression of TNF-alpha (16.5 pg/ml ± 3.2 pg/ml).

These findings suggest that co-culture of these primary cell types has a synergetic effect on the enhancement of TNF-α secretion. Its higher expression in the co-culture proves that the combination of both cell types is far more relevant to in vivo conditions, compared to primary ESC or EEC lines cultured separately. Furthermore culturing ESC and EEC together in the context of 3D culture conditions establishes an adequate architectural and functional in vitro model for studying of certain aspects of human implantation.



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M16FLU SEASON 2015/16 IN BULGARIA: ANTIGENIC AND MOLECULAR CHARACTERISTICS OF INFLUENZA VIRUSES

I. GEORGIEVA*, S. ANGELOVA, N. KORSUN

National Reference Laboratory „Influenza and ARD”, Department of Virology, National Centre of Infectious and Parasitic Diseases, Sofia, Bulgaria*Corresponding author: I. Georgieva, National Laboratory “Influenza and ARD”, Department of Virology, NCIPD, 44A Stoletov Blvd, 1233 Sofia, Bulgaria; E-mail: [email protected]; Tel/Fax: +359 29318132

Key words: Influenza viruses, antigenic, molecular, rRT-PCR

Influenza viruses cause annual epidemics during the winter season posing a significant threat for public health, therefore constant monitoring of genetic and antigenic properties of circulating viruses is very important. From week 40/2015 to week 20/2016 a total of 1127 nasopharyngeal swabs received from different regions of the country were tested. Influenza viruses were rRT-PCR confirmed in 318 (28%) of the cases. Influenza A type viruses have prevailed over the B type, and A(H1N1)pdm09 viruses greatly outnumbered A(H3N2) with 210 (87%) and 31 (13%) detections, respectively. All of the A(H1N1)pdm09 viruses characterized antigenically were similar to the vaccine virus A/California/7/2009. The Bulgarian A(H1N1)pdm09 strains characterized genetically fell into subclade 6B.1 according to sequences of HA and NA genes. Antigenic characterization of influenza A(H3N2) viruses by HI assay was hampered due to variable agglutination of red blood cells. Phylogenetic analysis of the HA genes of A(H3N2) viruses classified Bulgarian strains into 3C.2a subclade. Influenza B viruses detected in Bulgaria during the 15/16 flu season belonged to Victoria lineage, have HA genes that fall into the 1A clade and remain antigenically similar to vaccine virus B/Brisbane/60/2008.

Grant support: This study was funded by the Bulgarian Ministry of Health and National Influenza Pandemic Preparedness Plan.


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M17STUDYING THE ROLE OF ∆NP73 IN MITOCHONDRIAL FUNCTION

IRINA GEORGIEVA1, MARINA STANTIC2, MARGARETA WILHELM2

1 – Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria2 – Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden*Corresponding author: [email protected], MTC Department, Karolinska Institutet, Stockholm, Sweden

Keywords: breast cancer, TAp73, DNp73, OXPHOS genes, ROS accumulation

p73 is a member of the p53 protein family of transcription factors. The p73 gene has two alternative promoters. The most upstream promoter gives the full-length (TАp73) protein and the second internal promoter leads to the expression of the N-truncated (ΔNp73) isoform. The TAp73 acts as a tumor suppressor where the ΔNp73 isoform blocks the transcriptional activity of the full-length protein and thereby is believed to be an oncogene. High ΔNp73 protein levels have been observed in a number of tumors, including breast cancer where numerous studies also describe that mitochondrial function is impaired. RNA sequencing data from the TCGA database reviled that high levels of ΔNp73 in breast cancer patients correlated with downregulation of a number of key genes, implicated in mitochondrial metabolism. Here, we show that increase in ΔNp73 levels has a role in the deregulation of the mitochondrial function in breast cancer cell lines. Overexpression of ΔNp73 in MCF7 and MDA-MB-231 cell lines leads to decrease in the levels of the NRF1 gene, a transcriptional factor for subunits of the complexes in the respiratory chain. We show decrease expression of OXPHOS genes, found downregulated in breast cancer patient data set with high ΔNp73 levels. The dysfunction of the electron transport chain leads to the production of ROS, which we observed by performing flow cytometry analysis. It would be of clinical relevance to gain a deeper understanding of the metabolic switch, particularly mitochondrial bioenergetics deregulation, observed in the high ΔNp73 breast cancer patient data set.

Acknowledgment: This work was supported by the Amgen Foundation for the Amgen Scholars 2016 Program.


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M18PLANT GROWTH PROMOTING EFFECTS OF PSEUDOMONAS PUTIDA STRAINS

TEODORA GEORGIEVA*, YANA EVSTATIEVA, DILYANA NIKOLOVA, VALENTIN SAVOV

Department Biotechnology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: Biochemical characterization, siderophore, phosphate solubilisation, PGPR; Pseudomonas

The associated with the plant rhizosphere species of bacteria have beneficial effect on plant’s growth and crop yield and collectively called as PGPR. Pseudomonas species are one of the major constituents of rhizobacteria, encourage the plant growth through their diverse mechanisms and act as biocontrol agents for various agriculture plants. The bacterial strains are identified as Pseudomonas putida OR2, OR5 and Pseudomonas stutzeri, based on their complete 16S ribosomal gene sequencing. The last one is Pseudomonas putida BTCC1046 (CCM3423). The degradation of biochemical substrates by studied Pseudomonas strains is estimated by API 20NE (Biome´rieux, France) identification kit. In our study 3 strains Pseudomonas putida and a strain Pseudomonas stutzeri are screened for their plant growth promoting activity i.e. antifungal activity, siderophore production and phosphate solubilization. Studied activity was carried by growth on – CAS media for siderophore production and Pikovska agar plates for phosphate solubilisation. The antifungal activity is estimated on single layer agar method against Aspergillus flavus, Aspergillus niger, Fusarium graminearum, Penicillium claviforme and Rhizopus archizus. Pseudomonas putida OR5 and Pseudomonas putida BTCC1046 showed positive activity for siderophore production – respectively 40mm and 30mm diameter of discoloration area and phosphate solubilisation - respectively 15mm and 13 mm. The third strain P. putida OR2 and P. stutzeri did not confirmed positive PGPR activity. These pseudomonads did not suppress the growth of the tested fungi and more investigation about antifungal activity is needed.

Acknowledgment: This work was supported by the companies ROMB Ltd. and Advanced nutrients.


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M19RELATION BETWEEN MULTIPLE DEFECTS IN HUMAN SPERMATOZOA AND SPONTANEOUS ABORTIONS IN PATIENTS UNDERGOING IVF

VILYANA GEORGIEVA1, DIMITAR PARVANOV1, GUEORGUI STAMENOV1, TODOR CHAUSHEV1

1 – Nadezhda,Women’s Health Hospital, 3 “Blaga vest” Street, Sofia, Bulgaria*Corresponding author: Vilyana Georgieva, [email protected]

Keywords: multiple defects, human spermatozoa, spontaneous abortions, IVF/ICSI

To investigate the effect of the ‘male factor’ and especially the role of multiple defects in the pathogenesis of spontaneous abortion, 107 couples undergoing ICSI were studied for sperm parameters retrospectively. The patients were divided into two groups, depending on their reproductive outcome: (1) 65 couples who achieved a successful pregnancy (live births); (2) 42 couples who experienced spontaneous abortions. Couples with anovulation, tubal disease, endometriosis, polycystic ovarian syndrome and severe male factor infertility were excluded. Only cases with good quality oocytes were included. Evaluation of morphology was performed according to Tygerberg-Kruger strict criteria. Due to the retrospective nature of the study, the approval of our institutional board was not required. Statistical analysis was performed by using statistical program SPSS (version 21.0). Independent- Sample t-test was performed to find out whether any significant mean difference exist between the studied groups. Results were reported as mean ± standard deviation. Significant differences were considered when p-value is less than 0.05.

Significant differences in the percentage of found spermatozoa with multiple defects were observed between the two studied groups of patients. The mean value of multiple defects in the abortion group was higher compared to patient group with live births (40.26 ± 9.78 vs. 29.83 ± 7.24; P=0.047). An increased number of patients with spontaneous abortion in cases with elevated number of multiple sperm defects was observed. We found a dramatic increase of spontaneous abortions above a cut-off level of 40%. Miscarriage rate among the couples with more multiple defects in males than the mentioned cut-off was significantly higher (74%).

Patients with higher frequency of spermatozoa with multiple defects have an increased chance of spontaneous abortion. Thus, it is concluded that multiple defects found in human spermatozoa may serve as an essential morphological biomarker in the abortion risk assessment models.

Acknowledgment: This work was supported by Nadezhda,Women’s Health Hospital, Sofia, Bulgaria.


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M20DOES AGING AFFECTS AFFINITY OF PRYMERY STEMM CELLS AND OSTEOBLASTIC CELL LINE TO EXTRACELLULAR MATRIX PROTEINS?

KAMELIA HRISTOVA-PANUSHEVA1*, MILENA KEREMIDARSKA-MARKOVA1, GEORGE ALTANKOV2,3 AND NATALIA KRASTEVA1

1 Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, ”Acad. G. Bonchev” Str., bl. 21, 1113 Sofia, Bulgaria 2 Institute for Bioengineering of Catalonia, Barcelona, Spain and 3 ICREA*Corresponding author: [email protected]

Keywords: stem cells, senescence, tissue engineering, cell adhesion, cell morphology, matrix proteins

Mesenchymal stem cells (MSCs) have attracted much interest because of their self-renewing potential and multipotency for possible clinical use. Unlike embryonic stem cells adult stem cells are subject of aging processes and the concomitant decline in their function. Age-related changes in MSCs have to be well understood in order to develop clinical techniques and therapeutics based on these cells. In this work we have studied the effect of aging on adhesion behaviour of bone marrow-derived MSC and MG63 osteoblastic cells onto three matrix proteins: fibronectin (FN), vitronectin (VN) and collagen I (Coll I). Cell adhesion was assessed in terms of the degree of cell spreading, number of attached cells and overall cell morphology and the results ascertained a substantial difference among both cell types. Adhesive phenotype of bone marrow MSC was very dynamic in the first week of culture. At the 1st day the cell adhesion was highest on vitronectin, at the 3th day - on Coll I - and at the 7th day - on FN. With further incubation, up to the 21st day, the affinity of MSC to all studied proteins decreased. In contrast, MG63 cells demonstrated a stable cell adhesive phenotype with high affinity to FN and Coll I and low affinity to vitronectin over the whole culture period. These data suggest that adhesion behavior of MSCs to different proteins is affected from aging processes unlike MG63 cells and this age-related changes have to be consider when expanding adult stem cells for clinical applications.

Acknowledgment: This work was supported by Bulgarian National Scientific Research Fund, project №: DFNI_B01_18/2012.


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M21INTERACTION BETWEEN BACTERIA, KERATINOCYTES CELL LINE AND RGO, AG- RGO COLLAGEN NANOCOMPOSITES

NENKO IVANOV1, DRAGOMIRA STOYANOVA1, ANITA NIKOLOVA1, ALEKSANDRA HRISTOVA1, SILVIA YANEVA1, MONIKA BORISOVA1, SIMONA GALABOVA1, TODORKA VLADKOVA3, ILIANA IVANOVA1, ANELIAKOSTADINOVA2

1 – Department of Microbiology, Faculty of Biology, Sofia University “St. KlimentOhridski”, Sofia, Bulgaria2 –Institute of Biophysics and Biomedical Engineering, BAS, Sofia, Bulgaria3- University of Chemical Technology and Metallurgy, 8 “Kl. Ohridski” Blvd, 1756 Sofia, Bulgaria*Corresponding author: [email protected], Institute of Biophysics and Biomedical Engineering, BAS, acad.G.Bonchev 21, 1113 Sofia, Bulgaria

Keywords:collagennanocomposites, reduced graphene oxide, silver-reduced graphene oxide, cytotoxicity, antimicrobial effect

The aim of the present work was to establish the possibility of clinical application of some collagen nanocomposites with reduced graphene oxide and with or without silver nanoparticles. Several microbial strainsC. lusitaniae 74-4, S. enterica 2333 (WDCM 00029, DSMZ 4224), E. coli 3548 (ATCC 10536), P. putida 1090 (ATCC 10536), S. epidermidis 3486 (ATCC 10536) and Bacillus cereus 1095 (ATCC 11778), provided by the National Bank of Microorganisms and Cell Cultures (NBIMCC), Bulgaria, and keratinocyte line HaCaT were tested for the sensitivity to nanocomposites. Diffusion in solid nutrient medium was used to test antibacterial effect. Crystal violet stain was used for cytotoxicity determination and microscopic observation of cell morphology. It was established the stronger effect of the silver nanoparticles, reduced graphene oxide and collagen nanocomposite against tested microorganisms, but no effect on the keratinocytes cell line viability and survival. The obtained results demonstrate the possibility to apply such nanocomposite material in wound dressing, medical implants, catheters, drainages and other clinical applications.

Acknowledgment: This work was supported by COST Action TD1305 are gratefully acknowledged.


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M22PRESERVATION OF ACIDITHIOBACILLUS FERROOXIDANS JCM 3863 BY USING A LYOPHILIZED PROTECTIVE AGENT

DIMANA KABOVA1*, RALITSA VASSILEVA1, TANIA MITOVA1, YANA VASSILEVA1, VYARA MAMATARKOVA1

1 – Department of Biotechnology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected], Department of Biotechnology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, 8 Dragan Tzankov Blvd., 1164 Sofia, Bulgaria

Keywords: Acidithiobacillus ferrooxidans JCM 3863, biotechnology, microbiology, lyophilization

Bacteria Acidithiobacillus ferrooxidans are used in various biotechnological processes. All of these are based on the ability of these bacteria to oxidize ferrous to ferric ions. This process plays an important role in biohydrometallurgy, environmental biotechnology, waste gas treatment in particular, as well as in eliminating solid waste from industries containing heavy and toxic metal. Owing to the importance of At. ferrooxidans in fulfilling these tasks, a reliable method is required for a long-term preservation of valuable strains and in maintaining their effectiveness. Lyophilization shows excellent results in sustaining microorganisms in time. The purpouse of the experiment is to investigate the survival of At. ferrooxidans JCM 3863 after lyophilization. This research describes a process of lyophilization of the strain using new protective substances and the results of several rehydrations of the bacterial strain carried out in chronological intervals of time. After rehydration and cultivation on solid and liquid media the strain shows satisfactory survival.


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M23COMPARATIVE STUDY OF THE EFFECTS OF PLANT GROWTH REGULATORS AND ACTIVATED CHARCOAL ON THE DEVELOPMENTAL PATTERNS OF SIDERITIS SCARDICA IN VITRO CULTURES

PETYA KOLEVA1, INA ANEVA 2, LJUBA EVSTATIEVA2, KALINA DANOVA1*

1 – Institute of Organic Chemistry with Centre of Phytochemistry, BAS, Sofia, Bulgaria2 – Institute of Biodiversity and Ecosystem Research, BAS, Sofia, Bulgaria*Corresponding author: [email protected], Institute of Organic Chemistry with Centre of Phytochemistry, Acad. G. Bonchev 9 str, Sofia 1113, Bulgaria

Keywords: activated charcoal, growth and development, in vitro cultures, plant growth regulators, polyphenolics production, Sideritis scardica Griseb.

The Balkan endemic Sideritis scardica is traditionally utilized as a pulmonary treatment, as well as anti-flu and wound healing remedy. The low germination rate and collection pressure imposes significant risk on its natural populations. In a broader program for plant cell tissue and organ culture colelction establishment at the Institute of Organic Chemistry with Centre of Phytochemistry, shoot cultures of the species were initiated from surface sterilization of seeds of the plant, collected at peak Shabran in Slavianka mountain. Then, an array of plant growth regulators (PGRs) modifications, comprising of benzyl adenine (BA) and 1-naphtylacetyc acid (NAA) treatments, as well as different activated charcoal (AC) concentrations were applied. Their impact on developmental patterns and polyphenolics production were studied in vitro. As a general observation all treatments increased significantly the survival rate of explants, as well as the multiplication index and polyphenolics productions as compared with the non-treated control. A significant rise of FW accumulation was also recorded (much higher in PGR treatments, as compared with AC). The AC treatment was also observed to stimulate polyphenolics production and increase sub-culture period, thus improving explant quality to a higher extent as compared with PGR. Further investigations are in process to study in details the phytochemical profiles of the different treatments and scale-up the obtained processes to higher volume bioreactor system.

Acknowledgment: This work was supported by grant of the Swiss National Science Foundation and Ministry of Education and Science, Bulgaria, in the Framework in the Bulgarian-Swiss Research Program (BSRP, grant No. IZEBZO_142989; DO2-1153); PK - World Federation of Sciences – Bulgarian National Scholarship Programme for young researchers


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M24CULTIVATION OF SUSPENDED CELLS CULTURES AND BIOFILM OF ACIDITHIOBACILLUS FERROOXIDANS JCM 3863 IN PRESENCE OF COPPER IONS

KRISTINA LAZAROVA1*, VYARA MAMATARKOVA1

1 – Department of Biotechnology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, 1164 Sofia, 8 Dragan Tsankov blvd., Bulgaria*Corresponding author: [email protected], Department of Biotechnology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, 1164 Sofia, 8 Dragan Tsankov blvd., Bulgaria

Keywords: Acidithiobacillus ferrooxidans, biofilm, copper ions

Bacteria Acidithiobacillus ferrooxidans are implemented usually in many biotechnological processes as bioleaching of metals from low grade ores, purification of waste waters and gases, remove of metals from solid wastes. The bioprocess systems with biofilm of Аcidithiobacillus ferrooxidans are capable to oxidize ferrous to ferric ions in higher concentrations. The aim of this work is to investigate in different innicial concentration of ferrous ions their bacterial oxidation in submerged cultures and biofilm of Аcidithiobacillus ferrooxidans JCM 3863 in presence of copper ions ions. Submerged cultures and biofilm have been cultivated in shake flacks at 200 rpm in two media – with 9 and 12 g/l initial concentration of iron ions. The biofilm has been formed on inert carriers - polyethylene cylinders, four carriers in flasks. Biofilm and submerged cultures have been cultivated in five consecutive cycles in each nutrient medium in presence of 5 g/l copper ions to complete oxidation of ferrous to ferric ions. In the end the biofilm thickness has been determined. Mean rates of oxidation were compared for each cycle of cultivation. In consecutive cultivation of suspended cells in media 9K after the second cycle the mean rates decrease significant. In biofilm in media 9K is observed that the mean rates are constant and even increase in the last cycle. In consecutive cultivation of suspended cells in media 12K in the last two cycles is reached almost the rate from the firs cycle after decrease in the medium cycles. In biofilm in media 12K there is gradually increase of mean rates compared to the first cycle. In result of the made research can be made the conclusion that the biofilm is better about industrial biotechnological processes in presence of copper ions and high concentrations of ferrous ions.


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M25ANTIBACTERIAL AND ANTIFUNGAL ACTIVITY OF LACTOBACILLUS PLANTARUM STRAINS ISOLATED FROM BULGARIAN GREEN CHEESE

EMANUELA LUKACH1, DILYANA NIKOLOVA1, YANA EVSTATIEVA1, TEODORA GEORGIEVA1, DESSISLAVA DIMITROVA2, TEODORA IVANOVA2, ROSITSA TROPCHEVA1*

1 – Department of Biotechnology, Faculty of Biology, Sofia University St. Kliment Ohridski, 8, Dragan Tsankov Blvd., 1164, Sofia, Bulgaria.2 - Institute of Biodiversity and Ecosystem Research, Department of Plant and Fungal Diversity and Resources, Acad. Georgi Bonchev Str., bl. 23, 1113, Sofia, Bulgaria.*Corresponding author: [email protected], Department of Biotechnology, Faculty of Biology, Sofia University St. Kliment Ohridski, 8, Dragan Tsankov Blvd., 1164, Sofia, Bulgaria.

Keywords: Lactobacillus plantarum, antibacterial, antifungal activity, Bulgarian green cheese

Lactic acid bacteria are being increasingly promoted worldwide, assuming that they can play an important role in food quality and bio-preservation. The modern consumers evaluate not only the nutrition value of the products, but also their functional properties. Thus, a renovated interest on traditional fermented products has been observed. This study aimed to characterize the microbiota of the Bulgarian Green cheese (Cherni Vit cheese) and to estimate the beneficial properties of the lactic acid microflora of the product. The Green cheese is made from sheep milk and its characteristic green colour is due to the presence of moulds on the cheese’s crust. The isolated six strains were identified as representatives of the species Lactobacillus plantarum by 16S rRNA sequence analysis. In vitro antimicrobial activity of LAB, cultivated in MRS broth and milk was estimated. Tested L. plantarum strains possess a broad spectrum of activity against pathogenic bacteria (Escherichia coli, Staphylococcus aureus, Enterobacter feacalis, Pseudomonas aeruginosa, Pseudomonas putida, Bacillus subtilis). A complete inhibition against moulds from the species Aspergillus flavus, Aspergillus niger, Fusarium graminearum, Trichoderma viride and Penicillium claviforme, was demonstrated from all Lactobacillus plantarum strains. The presence of lactobacilli in the Green cheese adds value to the product’s functional qualities. The broad spectrum of antimicrobial activity of the isolated LAB is a promising advantage, suggesting their potential applications in different food technologies as a bio-preservative agent and a health promoting products against different pathogens.


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M26AN INFORMATION THEORETIC QUANTIFICATION OF CALCIUM SPECIFICITY

SYLVIA MARINOVA 1,2*, TEREZA VAZ MARTINS1, RICHARD MORRIS1

1 – Computational and Systems Biology Department, John Innes Centre, Norwich Research Park, Norwich, United Kingdom 2 – Faculty of Biology, University of Sofia “St. Kliment Ohridski”*Corresponding author: [email protected]

Keywords: calcium, specificity, plants, proteins, kinetics, information theory

Calcium is a ubiquitous signal in most eukaryotic organisms including plants. It is involved in a wide range of cellular processes ranging from cell proliferation to cell death and it plays pivotal role in plants response to environmental changes. A key question is how this single ion is responsible for controlling such a numerous cellular processes. It is well known that calcium signals exhibit characteristic temporal patterns known as calcium signatures which lead to specific downstream responses to different stimuli. To test whether calcium specificity is achieved due to a variation in enzyme kinetics of downstream calcium-sensitive proteins we used information theory as a well-known theoretical framework. Acknowledgment: This work was done as part of the International Undergraduate Summer School 2016 at John Innes Centre, Norwich Research Park


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M27VISUALIZATION OF HBEST-1 PROTEIN BY ATOMIC-FORCE MICROSCOPY

K. MLADENOVA1, S. PETROVA1 T. ANDREEVA2, N. MLADENOV3, V. MOSKOVA-DOUMANOVA1, T. TOPOUZOVA-HRISTOVA1, Y. PETSEVA1, K. BALASHEV4, Z. LALCHEV1, J. DOUMANOV1

1 - Sofia University “St. KlimentOhridski”, Faculty of Biology2 - Bulgarian Academy of Science, Institute of Biophysics and Biomedical Engineering3 - Medical University of Sofia, Faculty of Medicine4 - Sofia University “St. KlimentOhridski”, Faculty of Chemistry and pharmacy

Key words: bestrophin-1, atomic-force microscopy, protein morphology

Human bestrophin-1 (hBest-1) is a transmembrane channel protein, expressed in the basolateral membrane of retinal pigment epithelium (RPE) cells, astyrocytes and glia. It is a multifunctional protein that acts as a Ca2+-activated chloride channel, regulator of voltage-gated Ca2+ channels, a volume regulated Cl− channel, HCO3− channel, mediates glutamate release in astrocytes or gamma aminobutyric acid in glia. Mutations in the protein cause several diseases collectively referred as Bestrophinopathies Two models for the topology of hBest1in the plasma membrane have been proposed based onits amino acid sequence. Both models predict four transmembrane domains of the protein and both N- and C-termini are situated in the cytosol. Our study has been focused on visualizing of hBest1 in the presence of Ca2+ ions, Glu (glutamate) and GABA (gamma aminobutyric acid) on Langmuir-Blodget monolayerfilms by AFM (atomic-force microscopy). Pure hBest1 reveals significant surface activity and forms stable Langmuir monolayers at the air/buffer interface. This provide a possibility to Langmuir-Blodgett transfer of the hBest1 films from aqueous (150 mM NaCl, pH 7) to solid support and subsequent visualization by atomic-force microscopy. AFM images were performed with NanoScopeV system (Bruker Inc.) in tapping mode, immediately after transfer of the protein monolayer to a glass surface. By atomic-force microscopy high resolution we achieved visualization of the hBest1 and indicated that Ca2+ ions cause oligomerization, molecular and macro-organization in protein films.

Acknowledgments: This work was supported by grant № 93/12.04.2016 from Sofia University.


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M28RESPONSE OF DNA METHYLATION MUTANTS OF ARABIDOPSIS THALIANA TO HIGH SALINITY

VELLA NIKOLOVA1*, VALYA VASSILEVA2

1 - Faculty of Biology - Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria2 - Institute of Plant Physiology and Genetics - Bulgarian Academy of Science, Sofia, Bulgaria*Corresponding author: [email protected], Faculty of Biology – Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria

Keywords: DNA methylation, salinity, halotropism, gene expression, root growth, Arabidopsis thaliana

Salinity stress affects almost every aspect of plant growth and development. Plants can alleviate salt stress effects through halotropism, an adaptive growth of plant roots in which they bend away from salt. In this study, the response to high salinity of two Arabidopsis thaliana mutants, met1 and ddm1, which have a global loss of genome CG methylation marks, was examined. The wild-type Col-0 and ddm1 seedlings showed a halotropic response, whereas met1 seedlings exhibited no change in their growth direction and followed the gravity vector. High salinity led to changes in expression of the genes from the Salt sensitive (SOS) pathway, which could be induced by a change in genome DNA methylation status. The reduced methylation in met1 correlated with a higher expression of the tested SOS genes, which was exactly opposite for ddm1 and Col-0. Their methylation status and expression levels did not change. Thus, we demonstrate that proper DNA methylation is essential for the ability of roots to optimize their growth direction in stress environment.

Acknowledgement: This work was partially supported by the Marie Curie Intra European Fellowship to V.V. (Grant No. 326711, FP7-PEOPLE-2012-IEF) and the project BG05M2OP001-2.002-0001 „Studentski praktiki“


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M29HIGHER FREQUENCY OF TAIL DEFECTS IN HUMAN SPERMATOZOA CORRESPOND TO HIGHER LEVELS OF DNA FRAGMENTATION

DIMITAR PARVANOV1, VILYANA GEORGIEVA1,GUEORGUI STAMENOV1, TODOR CHAUSHEV1

1 – Nadezhda,Women’s Health Hospital, 3 “Blaga vest” Street, Sofia, Bulgaria*Corresponding author: Dimitar Parvanov, [email protected]

Keywords: human spermatozoa, tail defects, DNA fragmentation

Numerous studies have demonstrated the presence of increased levels of sperm DNA fragmentation in patients with poor sperm morphology. The majority of studies have shown that specific head abnormalities (long, amorphous and micro heads) are associated with the presence of spermatozoa with immature chromatin and higher DFI (DNA fragmentation index). During spermatogenesis the process of chromatin compactization is considered to be tightly linked to the head morphology. However, it could also be suggested that there are relations between DNA integrity and other morphological defects. A retrospective study was performed during the period 2015 - 2016. A native sperm from 98 patients undergoing ICSI procedure were used for analysis of DNA fragmentation and sperm morphology. Using the Sperm Chromatin Structure Assay (SCSA), the number of DNA fragmented sperm, expressed as DNA fragmentation index (DFI) was evaluated. Sperm morphology was assessed by Kruger strict criteria after staining procedure with Sperm Stain kit (Microptic). Spearman’s rank correlation test was performed to evaluate the correlation between the frequency of the morphological defects and levels of DNA fragmentation. Among all types of morphological defects, tail defects showed the strongest correlation with DNA fragmentation of human spermatozoa. A significant positive correlation was observed between DFI and percentage of cumulative tail defects (r = 0.54; p < 0.001), absent tail (r = 0.46; p < 0.001), short tail (r = 0.37; p = 0.022) and coiled tail (r = 0.34; p = 0.015). The results obtained lead to a better understanding of the reasons for the existing levels of DNA fragmentation in human spermatozoa. Considering the crucial importance of cumulative and particular tail defects, the frequency of their occurrence could serve as a useful indicator of sperm DNA integrity.



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M30COMPARATIVE INVESTIGATION OF THE MODERATE HEAT STRESS REACTION BETWEEN TWO ECOTYPES OF PLANE (PLATANAUS ORIENTALIS) AND OLIVE (OLEA EUROPAEA) TREES

M. PAUNOV1, B. PAVLOVA1, E. CHACHI2, ST. DIMITROVA1, K. DANKOV1, V. VELIKOVA3, TS. TSONEV3, M. KOUZMANOVA1, V. GOLTSEV1

1 - Department of Biophysics and Radiology, Faculty of Biology, St. Kl. Ohridski University of Sofia, 8 Dragan Tzankov Blvd., Sofia, 1164, Bulgaria2 - Department of Physics, Faculty of Technical Sciences, I. Qemali University of Vlora, Albania3 - Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Acad. G. Bontchev Str., bl. 21,1113 Sofia, Bulgaria

Plane (Platanus orientalis) and olive (Olea europaea) both grow in a high temperature region of the east Mediterranean. Both species have economic importance for the countries in the region, being used for ornamental or edible purposes. Temperature heavily impacts photosynthesis, thus the whole productivity of the plants. Moderate temperature changes (in the range of 15°C) occur daily in the nature and induce moderate stress responses in the plants. The photosynthetic electron transport chain is highly sensitive to stress and its reactions can be probed by chlorophyll a fluorescence measurements. Analysis of those measurements can be done by the method called “JIP test”, giving whole range of parameters informative for the photosynthetic performance and characterizing the changes during and after stress. In this study a comparison between plane (Platanus orientalis) and olive (Olea europaea) was conducted to investigate the differences and similarities in the response of their photosynthetic machinery to moderate temperature stress. Two years old plane trees developed from seeds of Bulgarian and Italian areas and three years old olive tree grown in Albania were used. The temperature was raised for 3 days from the daily normal of 28°C step by step to 41°C where it stood for 4 hours before returning to the normal. Consequently a 3 day recovery period was applied. We observed that “Bulgarian” planes responded slightly negative or even positively to the temperature rise while “Italian” plane and olive reacted with a considerable decrease in the photosynthetic performance. We speculate that this observation could be connected to the climate where those plants or their parents grow: Italian and Albanian habitats are in the Mediterranean climate zone while Bulgarian habitat is on the border of Mediterranean with the moderate climate zone. However, while the leaves of both planes showed recovery in their photosynthetic parameters during the recovery period the olive leaves did not.

Acknowledgment: We are grateful to Bulgarian National Scientific Fund (Project № DFNI B02/8) for financial support.


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M31INTRACELLULER SURVIVAL OF PSEUDOMONAS AERUGINOSA PAO1 IN A549 CELLS

TSVETELINA PAUNOVA-KRASTEVA1, VASILKA VELKOVA 2, DAYANA BORISOVA1,

STOYANKA STOITSOVA1

1 – Department of General Microbiology, Institute of Microbiology, BAS, Sofia, Bulgaria2 – Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected], Department of General Microbiology, Institute of Microbiology, BAS, 26 Georgi Bonchev str., Sofia 1113, Bulgaria, Tel. +359 2 979 31 59

Keywords: Pseudomonas aeruginosa, human lung carcinoma A549 cells, GFP

In their interaction with host epithelia, different strains of Pseudomonas aeruginosa manifest different phenotypes: they may remain adherent to the surface of the eukaryotic cells (adherent phenotype), or penetrate and survive intracellularly (invasive phenotype). The aim of the study is to characterize this aspect of the interaction of strain P. aeruginosa PAO1 with human lung carcinoma A549 cells. Initially, we performed a co-cultivation test followed by determination of the number of intracellular bacteria by plating on agar. The result indicated that the bacterium is of the invasive phenotype. Using a three-parental labeling system (including E. coli MT 102 pBAH 8 as donor, and E. coli HB 101 pR K600 as helper) the strain was labeled with green fluorescent protein. Fluorescent microscopy of the GFP-PAO1 strain after co-cultivation with A549 cells confirmed the intracellular survival of the bacteria.

Acknowledgment: Acknowledgement: This work was supported by “Program for career development of young scientists”, BAS, Contract № DFNP-55/27.04.2016


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M32EFFECT OF SNAKE VENOM PHOSPHOLIPASE A2 ON ACTIN CYTOSKELETON REARRANGEMENT OF RETINAL PIGMENT EPITHELIUM CELL LINES

YULIA PETSEVA1, KIRILKA MLADENOVA1, STOYANKA STOITSOVA2, JORDAN DOUMANOV1AND SVETLA PETROVA1

1 - Sofia University “St. Kliment Ohridski”, Faculty of Biology2 - Bulgarian Academy of Sciences, Institute of MicrobiologyCorresponding author: [email protected]

Key words: sPLA2, RPE, actin cytoskeleton, transepithelial resistance

Snake venoms are complex and multifunctional mixtures of proteins, polypeptides and other bioactive molecules, used as important molecular tools to understand in details various biological processes. The main neurotoxin of Vipera ammodytes meriodionalis snake venom–Vipoxin, is a heterodimeric protein composed of a neurotoxic phospholipase A2(PLA2) subunit and a non-toxic and catalytically non-active one, associated in a tight complex. Secreted PLA2 (sPLA2, EC 3.1.1.4) catalyse the hydrolysis of the 2-acyl ester bond of 1,2-diacyl-3-sn-phosphoglycerides in a calcium-dependent manner, releasing as products lysophospholipids and free fatty acids - potent lipid mediators with significant effect on different cell structures and functions. Our study has been focused on cell effects of the purified toxic sPLA2 vipoxin subunit on cell integrity and survival of two human retinal pigment epithelium (RPE) cell lines - RPE-1 and ARPE-19 as RPE cells play a key role in photoreceptor homeostasis and are strongly affected by the action of various pharmacologically active agents. The effects of different concentrations of toxic sPLA2 on actin rearrangement of RPE-1 and ARPE-19 cells were detected using actin fluorescence staining. Measuring of cell transepithelial resistance was applied for assessing the permeability of tight junctions and/or membrane perturbations. Our results showed a considerable rearrangement of actin cytoskeleton and altered morphology of the cells induced by catalytic/pharmacological activity of toxics PLA2 vipoxin subunit.

Acknowledgments This work was supported by grants: № 93 12/04/2016, Sofia University


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M33EX SITU CONSERVATION AND BIOLOGICALY ACTIVE COMPOUDS IN ACHILLEA THRACICA

MARIYA ROGOVA1, DETELIN STEFANOV2, ANTONIYA PARVANOVA1, VENETA KAPCHINA-TOTEVA1

1 – Department of Plant Physiology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria2 – Department of Biophysics, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected], Department of Plant Physiology, Faculty of Biology, Sofia University "St. Kliment Ohridski", 1164, Dragan Tzankov blv. № 8, Sofia, Bulgaria

Keywords: Achillea thracica, ex situ conservation, photosynthesis, biologically active compounds

The genus Achillea (fam. Asteraceae) is presented globally by 130 species, with 19 naturally distributed species in Bulgaria. Achillea thracica Velen., is a Bulgarian endemic medical plant under the protection of the Bulgarian Biodiversity Law (2002) and is included in the Red Data Book of Bulgaria with the national conservation status: critically endangered (CR). The species is in the Bern Convention on the Conservation of the European Wildlife and Natural Habitats as “rare” (1979) and in the IUCN’s European Red List of Vascular Plants as “Data Deficient” (DD). The present work aims to assess the development of plants on their way to successful ex situ conservation. Intact plant material of Achillea thracica Velen. was collected at its natural habitat near Manole village, Plovdiv, Bulgaria. In vitro shoot cultures were induced from ripe dry seeds, collected from in vivo growing wild plant and sterilized with 70% ethanol. Regenerated plants were propagated under aseptic controlled environmental conditions, on basal MS-B5 (Murashige and Skoog medium supplied with Gamburg B5 vitamins) medium containing 3% (w/v) sucrose and 0.7g.l-1 agar without any supplement of growth regulators. 100% of the ex vitro adapted plants were survived. Analyses of photosynthetic activity and the photosynthetic pigments were done. Comparative analysis of phenol and flavonoid compounds and antioxidant activities of extracts of A. thracica plants also were done. The results suggested that the in vitro growth conditions affect the accumulation of the plastid pigments, the secondary metabolites and antioxidant activities.

Acknowledgment: This work was financially supported by the grand 141/2016 Sofia University, Bulgaria.


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M34THE COMPARISON BETWEEN MAGNESIUM DEFICIENCY AND SULPHUR DEFICIENCY FOR EARLY DETECTION OF CHLOROPHYLL FLUORESCENCE IN RADISH PLANTS

IZABELA A. SAMBORSKA1, VASILIJ GOLTSEV2, LESZEK SIECZKO3, HAZEM M. KALAJI1

1 - Department of Plant Physiology, Warsaw University of Life Sciences SGGW, 02-776 Warsaw, Poland2 - Department of Biophysics and Radiobiology, Faculty of Biology, St. Kl. Ohridski University of Sofia, 8 Dragan Tzankov Blvd., Sofia, 1164, Bulgaria3 - Department of Experimental Statistics and Bioinformatics, Faculty of Agriculture and Biology, Warsaw University of Life Sciences WULS- SGGW, 02-776, Warsaw, Poland

Magnesium plays molecular and physiological roles in plants, as it is a component of the chlorophyll molecule, a structural stabilizer for various nucleotides and a cofactor for many processes associated with phosphorylation and dephosphorylation. Sulphur is one of the most difficult mineral deficiency to detect in plants. It takes part in many amino acides metabolism pathways. Sulfur is a vital part of all plant proteins, and certain plant hormones. Sulfur is mobile macronutrient within the plant, therefore deficiency symptoms usually start on the younger leaves and progress over time to the older leaves Chlorophyll fluorescence signals have been used mainly for detection and monitoring of the effect of different stressors on the performance of photosynthesizing organisms. Chlorophyll fluorescence measurments were down on radish plants with Hoagland solution in two sets of conditions, in macronutrient deficient solution (magnesium or sulphur) and full nutrition solution in phytotron. The photosynthetic efficiency of plants is evaluated by the use of an innovative method involving a combination of measurements of chlorophyll fluorescence signals, their detailed analysis, using the JIP test. Our experiments show that magnesium deficiency and sulphur deficiency have other mechanism in radish plants and both of them are difficult to observed by naked eyes. Usually symptoms of magnesium deficiency and sulphur deficiency are visible when plants are in bad conditions and it can be sometimes to late. That is why detailed analysis of chlorophyll fluorescence measurments and JIP test may be helped in diagnosis this two deficiencies.


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M35PROTEASOME INHIBITION CAN MODIFY THE LEVELS OF INFLAMMATORY CYTOKINES IN IRRADIATED BLOOD MONONUCLEAR CELLS

GERGANA SAVOVA*, NEVENA ANEVA, KATYA STANKOVA, RAYNA BOTEVA

Department of Molecular radiobiology, Nacional Centre of Radiobiology and Radiation Protection, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: proteasome, proinflammatory cytokines, NF-kB, ionizing radiation

The proteasome is a multicatalytic enzyme complex located in the cytoplasm and nucleus of all eukaryotic cells. Its primary function is to degrade damaged and misfolded proteins. The proteasome is also actively involved in the cell cycle control, apoptosis, regulation of transcription and radiation response. It has been shown that its inhibition can induce changes in the radiosensitivity among various cell types. In response to different stimuli, including ionizing radiation, cells produce inflammatory cytokines, which can induce inflammation. The current study aims to investigate the effect of proteasome inhibition by the peptide aldehyde MG132 on the levels of the pro-inflammatory cytokines-tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-8 (IL-8) in irradiated human peripheral blood mononuclear cells (PBMCs). Our results showed that MG132 can decrease significantly the levels of the three cytokines measured 24 hours post-irradiation. As the NF-kB is actively involved in cytokine transcription pathway, our results are consistent with inhibition of NF-kB upon proteasome inhibition by MG132, which suggests important role of the proteasome in the regulation of inflammatory response in irradiated PBMCs.


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M36POTENTIAL USE OF PLANT-GROWTH PROMOTING BACTERIA STRAINS FOR SELECTIVE BIOCONTROL

DMITRII SEMENOVSKII, YANA EVSTATIEVA, DILYANA NIKOLOVA, TEODORA GEORGIEVA, VALENTIN SAVOV

Department of Biotechnology Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: Plant-growth-promoting rhizobacteria; Antifungal agent; Biocontrol

Fungal disease causes noticeable expenses in agriculture. Reducing of these expenses is one of the most important problems, and that’s why disease control is needed. It is well known now that some bacteria and fungi are able to aggressively colonise soil and the roots of plants, also they are able to protect the plants roots from infections by soilborn pathogenic fungus. This phenomenon, named biocontrol, is caused by production of selective antifungal compounds, and non-selective acting compounds which affects plants and bacteria by modifying the chemical composition of the soil. In present study we have investigated antifungal activity of several PGPR strains (Bacillus subtilis and Bacillus amyloliquefaciens) isolated from Bulgarian soils. We have investigated this activity in vitro against the list of pathogenic fungus, which is commonly found in Bulgarian soils. Also we executed several tests to check their biochemical activity. We investigated bacteria-plant interaction through the experiment in vivo with plant seeds. It was founded noticeable antagonistic effect against Aspergillus niger, Aspergillus flavus, Fusarium graminearum, Penicillum claviforme, Rhizopus arhisus and Trichoderma viride which are varied between the PGPR strains. In the experiment with plant seeds it was founded noticeable plant growth modulating effect which depends on the used concentration of supernatant of the cultural media. Also there was observed dependence between growth modulating effect and growth phase of the bacteria culture. The investigated strains show great potential for use as biocontrol agents.


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M37EFFECT OF CYTOKININS AND AUXINS ON GROWTH AND ANTIOXIDANT ACTIVITY IN IN VITRO CULTURED EXACUM AFFINE L.

KATERINA SERAFIMOVA1, SYLVIA MARINOVA1, IRINA GEORGIEVA1, ANI KERCHEVA1, DESISLAVA POPOVA1, MIROSLAVA ZHIPONOVA*2, GANKA CHANEVA2, VENETA M. KAPCHINA-TOTEVA*2

1 – Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria2 – Department of Plant Physiology, Faculty of Biology, University of Sofia, 8 Dragan Tsankov Blvd., 1164 Sofia, Bulgaria; 2AgroBioInstitute, 8 Dragan Tsankov Blvd., 1164 Sofia, Bulgaria*Corresponding author: [email protected], Department of Plant Physiology, Faculty of Biology, University of Sofia, 8 Dragan Tsankov Blvd., 1164 Sofia, Bulgaria; 2AgroBioInstitute, 8 Dragan Tsankov Blvd., 1164 Sofia, Bulgaria

Keywords: cytokinins, auxins, Exacum affine, micropropagation, peroxidase

Exacum affine, commonly called Persian violet, is a member of the Gentianaceae family and it is grown for its beautiful flowers as a pot plant. The plant is susceptible to pathogens and the micropropagation approach provides as advantage the rapid obtaining of a large number of disease-free cuttings. In order to improve the growth of E. affine and its further ex vitro adaptation, we tested the effect of growth regulators, such as the plant hormones auxin (naphthaleneacetic acid, NAA) and cytokinin (benzylaminopurine, BAP) on morphometric parameters of in vitro grown plants, as well as on their antioxidant potential (non-enzymatic and enzymatic). Our data indicated that NAA can be applied to promote shoot elongation and root formation, while BAP leads to increase in the shoot number. The promotion of growth by both hormones correlates with lack of oxidative stress as indicated by the measurement of total antioxidant activity, total content of phenolic compounds and the peroxidase (POD) enzyme activity. Based on the data, we can conclude that the studied plant hormones can be used to modulate the growth of E. affine. However, the period of treatment and the applied concentrations should be further optimized.


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M38PREIMPLANTATION GENETIC DIAGNOSIS FOR MOSAIC FRAGILE CHROMOSOME 16

SLAVYANA YANEVA STAYKOVA1,2, RADA STANEVA1,2, BLAGA RUKOVA1,2, KRISTINA NIKOLOVA2, DRAGA TONCHEVA1, GUEORGUI STAMENOV2, SAVINA HADJIDEKOVA1,2

1Department of Medical Genetics, Faculty of Medicine, Medical University - Sofia2Nadezhda Women’s Health Hospital*Corresponding author: Slavyana Yaneva Staykova, [email protected]

Keywords: Whole-genome array-based comparative genomic hybridization (array CGH), preimplantation genetic diagnosis, miscarriage, spontaneous abortion, mosaic fragile chromosome 16.

About 10% to 15% of all clinically recognized pregnancies end in spontaneous abortion, most of them occurring during the first trimester. The cause for the vast majority of them are chromosomal abnormalities - de novo aneuploidy or polyploidy, de novo or inherited unbalanced chromosomal rearrangements. A little over 50% of missed abortions, as well as 7% of perinatal mortality cases and a plethora of other reproductive failures are reported to be due to chromosomal abnormalities. The microarray-based comparative genomic hybridization method, developed in the late 90s, provides high resolution and sensitivity and thus is currently used as a golden standard for pre-implantation genetic diagnosis of chromosomal aberrations. Array CGH allows identification of new microstructural aberrations and detection of chromosomal imbalances in seemingly balanced rearrangements. The application of PGA (preimplantation genetic analysis) in families with chromosomal aberrations reduces the risk of miscarriage, perinatal mortality or birth of children with chromosomal disorders. We present a couple with reproductive failures that suffered three clinical miscarriages in the first trimester and two ectopic pregnancies. The woman also described irregular menstrual cycle in the past two years. A hysterosalpingography was performed and complete occlusion of the fallopian tubes was established. The female patient was diagnosed with tubar factor infertility, chronic salpingitis and oophoritis. The following two years, the couple underwent ten unsuccessful IVF treatments. The female patient was additionally suffering from chronic ulcerative colitis, hypothyroidism, iron deficiency anemia and essential hypertension. Karyotyping was assigned to both partners. The cytogenetic analysis of the female patient revealed a fragile site on chromosome 16 in 10% of the analyzed cells – mos 46,XX,fra(16)(q22)[10]/46,XX[90]. This chromosomal finding was confirmed by retesting six months after the initial investigation. Such chromosomal abnormalities increase the risk for formation of unbalanced embryos. In order to select balanced/normal embryos and avoid future reproductive failures the couple underwent preimplantation genetic analysis. Five-day embryos were bopised and five to seven trophectoderm cells were obtained for genetic analysis. Cell DNA was amplified by SurePlex protocol (Illumina). Whole-genome array-based comparative genomic hybridization (array CGH) was performed using the BlueGnome 24sure + arrays, version 1.0 (Illumina) according to manufacturer's protocol. Arrays were scanned at 532 nm (Cy3) and 635 nm (Cy5) using Agilent’s High-Resolution C Scanner G2505C. The images were analyzed by BlueFuse


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Multi software, version 4.2. (Illumina). Eight embryos collected in the course of three IVF cycles were analyzed, two of which had balanced/normal karyotype and six were unbalanced. After frozen embryo transfer of both normal embryos, a singleton pregnancy was achieved that resulted in the birth of a healthy child. This case shows the advantages of whole genome microarray analysis as PGA in couples, where one or both partners are carriers of chromosomal aberrations. PGA decreases the incidence of recurrent pregnancy loss, increases the chance of balanced embryo implantation and the birth of a healthy child.

Acknowledgment: This work was supported by Nadezhda,Women’s Health Hospital, Sofia, Bulgaria.

M39ANTIOXIDANT RESPONSE IN CONDITIONS OF SALT STRESS OF THREE CEREAL CROPS OF THE GENUS TRITICUM

DENITSA TEOFANOVA1*, LYUBEN ZAGORCHEV1, KRISTIYANA MARINOVA1, IVANELA ALBANOVA1, MARIELA ODJAKOVA1

1 – Department of Biochemistry, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected], Department of Biochemistry, Faculty of Biology, Sofia University “St. Kliment Ohridski”, 8 Dragan Tsankov blvd., 1164, Sofia, Bulgaria

Keywords: ancient cereals, antioxidant enzymes, chlorophyll, hydrogen peroxide, L-proline

Salinity is among the most significant abiotic stresses, causing substantial yield losses in agriculture worldwide. Therefore, the characterization of the relative salt tolerance of major crop plants and identification of cultivars, suitable for cultivation in salinized arable lands is essential for the contemporary agricultural practice. Several alternative, referred as ancient wheat relatives, fall into the recent focus of research because of their supposedly better nutritional quality and resistance to unfavourable environmental conditions. Two ancient crops, kamut (Triticum turanicum) and spelt (Triticum spelt) were compared to their close relative, common wheat (Triticum aestivum) in their salt stress response in the present study. Various biochemical markers such as hydrogen peroxide, L-proline, chlorophyll a and b and antioxidant enzymes, namely glutathione reductase, superoxide dismutase, catalase, peroxidase and ascorbate peroxidase were compared at 0.1 and 0.3 M NaCl treatment after 14 days. All of them were differentially affected by the two salt concentrations, indicating relative adaptation to the lower salt stress and severe inhibition of metabolic functions at the higher salt. The response was similar in the three crop species, showing minor variations and suggesting that kamut and spelt were less affected by 0.3 M NaCl, compared to wheat.

Acknowledgment: This work was supported by fellowship award from L’Oréal-UNESCO for Women in Science’s national program, Bulgaria.


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M40ANTIFUNGAL POTENTIAL OF SOME COLLAGEN-BASED NANOCOMPOSITES AGAINST CANDIDA LUSITANIAE

DRAGOMIRA STOYANOVA1*, ILIANA IVANOVA1, TODORKA VLADKOVA2

1-Department of General and Industrial Microbiology, Sofia University, “St.Kliment Ohridski”, Bulgaria2-Polymer department, University of Chemical Technology and Metallurgy. Sofia. Bulgaria;*Corresponding author: [email protected]

Key words: pathogenic fungi, collagen matrix, nanoparticles, zone of inhibition

In this article the potential and a possible mechanism of action of different nanomaterials on pathogenic fungi, significant for medical practice is summarized. Furthermore, the antifungal action of different collagen-based nanocomposites against Candida lusitaniae is evaluated and compared. Collagen-based biomaterials with antimicrobial activity are attractive candidates for wound dressing, tissue engineering, components of implantable devices, etc. One of the easiest and most effective ways among the large variety of known approaches to add antimicrobial activity of biomaterials is development of composites including antimicrobial agents. In this study RGO, Ag/RGO, Ag/SiO/RGO, ZnTiO3, ZnTiO

3/SiO2/RGO and Ag/ZnO/ZnTiO3

respectively were entrapped in a porous collagen matrix by sol-gel cryogen drying to preserve the native biological activity of the collagen. No chemical interactions were expected in the Collagen/ZnTiO3 (or the other compounds) nanocomposites under these conditions. The nanocomposites were prepared in different ratios of the compounds. The formed sterile zones around the disc samples (diameter of 9.0 mm; thickness of 3 mm) were measured in mm (±0.5). The representative of eukaryotic organisms Candida lusitaniaе demonstrated high sensitivity with large sterile areas around some of the tested materials.

Acknowledgments: This study was conducted with the assistance of the Department of General and Industrial Microbiology in Faculty of Biology at Sofia University “St. Kliment Ohridski”. Bulgarian Scientific Fund is gratefully acknowledged for the financial support of this investigation (Grant DKOST 01/14/16.08.2016.)


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M41ANALYSIS OF THE PHILOGENETIC RELATEDNESS OF BALKAN TROUT SPECIMENS (SALMO TRURRA M. FARIO) FROM DIFFERENT RIVERS ON THE TERRITORY OF SOUTH-WEST BULGARIA

JOE DAVIS VELCHEV1, ROSSICA MARINOVA2, MARIA DIMITROVA1, GALERIDA RAIKOVA2, SVETOSLAV G. DIMOV1*

1 – Department of Genetics, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria2 – Department of General and Applied Hydrobiology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected], Sofia University “St. Kliment Ohridski”, Faculty of Biology, Department of Genetics, 1164 Sofia, 8 Dragan Tzankov Blvd, Bulgaria

Keywords: Salmo trutta m. fario; molecular phylogeny

To accomplish partial characterisation of the population of brown trout (Salmo trutta m. fario) in the Bulgarian mountain “Rila”, seventy individuals were analysed. The first part of the analysis included two direct PCRs using two pairs of primers amplifying the D-loop region of the mitochondrial genome, as well as a pair of primers that target the “housekeeping” gene for the lactate dehydrogenase enzyme. The resulting products of the PCRs were separately subjected to RFLP analyses with two different restriction enzymes. Twenty-six polymorphic profiles were obtained, each one of them comprising from 1 to 9 individuals. UPGMA analyses were used to construct a phylogenetic trees based on the D-loop region sequencing. The results were plotted to the rivers from which the samples were gathered. We established a clear tendency of grouping of some of the polymorphic profiles by the river from which the individuals were sampled, although some genetically identical individuals regarding the examined characteristics were also found in different rivers. In most of the rivers there were individuals with more than one genotype. On the territory of National Park “Rila” we discovered individuals that carry non-autochthonous traits, which is a clear testimony that the rivers have been restocked with salmons that have Atlantic traits. After looking into the individuals from rivers which are closely positioned to the ones that had been restocked we confirmed our apprehensions that there had been a genetic interchange between native and non-native individuals, which might result in the disappearance of the Balkan line of salmons.

Acknowledgment: This work was supported by the “Balkanka” Association’s project “Encouragement of Collaboration between NGOs and State Structures to Restore Native Trout in Rila Mountain”


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M42COMPARATIVE ANALYSIS OF CYTOTOXICITY OF CHLOROFORM EXTRACTS FROM LAMIUM ALBUM L. AND MORPHOLOGICAL ALTERATIONS IN MAMMALIAN CELLS

RALITSA VELEVA1, VESELINA MOSKOVA-DOUMANOVA 1, DANIELA DRAGOLOVA2, JORDAN DOUMANOV3, TANYA TOPOUZOVA-HRISTOVA1*

1 – Department of Cytology, Histology and Embryology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria2 – Department of Plant Physiology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria 3 – Department of Biochemistry, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected], Faculty of Biology, SU “St. Kliment Ohridski”, Sofia, Bulgaria

Keywords: Lamium album L., cytotoxicity, chloroform extracts

Lamium album is plant commonly used in traditional medicine because of its anti-inflammatory, astringent, antiseptic and antispasmodic activities. There are differences in the composition of secondary metabolites in plants grown in their natural habitats (in vivo), micropropagated plants (in vitro) and after their subsequent cultivation in a natural environment (ex vitro). This amendment leads to alteration in their biological activity. In this study we aim to compare the activity of chloroform Soxhlet extracts from Lamium album L. cultivated diversely. We have used A549 human cancer cell line and normal kidney epithelial cells MDCKII. We have performed cell cytotoxicity tests (crystal violet assay, MTT assay) and membrane permeability test (trypan blue exclusion assay) as well as morphological analysis of changes induced by the extracts. Upon treatment with extract ex vitro has been observed a large percentage of cells with damaged membranes at 24 hours. After 48 hours the total number of cells has been reduced, but the cells with damaged membranes have a relatively small rate. For comparison, 48 hours after treatment with in vitro extract, 100% damaged A549 cells has been observed.


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E1FUNGICIDE AMENDMENT OF AGRICULTURAL SOILS SELECTS ANTIBIOTIC RESISTANT BACTERIA

MICHAELA ALEXOVA1,2*, SILVENA BOTEVA1, ANELIA KENAROVA1, GALINA RADEVA2

1 – Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria2 – Roumen Tsanev Institute of Molecular Biology, Bulgarian Academy of Sciences, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: antibiotic, resistance, fungicide, bacteria, microcosms

The spread of resistant bacteria and antibiotic resistance genes in all environments led to human and animal health consequences. The aim of this study was to investigate the dose-response relationship between azoxystrobin applied to soils with different textures and the antibiotic resistance of indigenous bacteria. The occurrence of antibiotic resistance was followed in microcosms with increasing fungicide’s concentrations during three months of soil incubation at steady laboratory conditions. The antibiotic resistance of soil cell extracts against seven antibiotics belonging to aminoglycosides, tetracyclines, amphenicols, and penicillins was tested. Increased antibiotic resistance was dependent on the fungicide concentration, time of soil incubation and the antibiotic chemical structure. The time of transformations from sensitive to resistant bacterial strains was antibiotic dependent- short for ampicillin and tetracycline and long for spectinomycin and kanamycin. The higher fungicide concentrations induced higher antibiotic resistance in bacteria. More evident was the induced antibiotic resistance in sandy than in clay soils. Based on the results, it was concluded that fungicide pollution can be interpreted as a selective factor on antibiotic resistance of soil bacteria.


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E2FUNCTIONAL CHARACTERISTICS OF PHOTOSYNTHETIC APPARATUS IN HALOPHYTE PLANT, ERYNGIUM MARITIMUM L.

LILIA ANGELOVA1*, ALBENA IVANOVA1, SVETLANA MOMCHILOVA², ALBENA MOMCHILOVA3, LILIANA MASLENKOVA3

1 –Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Sofia 1113, Bulgaria2 – Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences3 – Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Sofia 1113, Bulgaria*Corresponding author: [email protected], Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Acad. Georgi Bonchev Str., Block 21, Sofia 1113, Bulgaria

Keywords: Eryngium maritimum, oxygen evolution, Photosystem II activity, thermoluminescence

Eryngium maritimum L. (Apiaceae) is one of the most salt-tolerant coastal dune plant, being therefore excellent model to study salt resistance and tolerance mechanisms. Halophytic strategies to salinity goes from simple morphological adjustments and production of specific osmolytes to the induction of specific mechanisms for efficient energy dissipation in order to prevent the accumulation of excessive energy within the chloroplasts and to avoid the production of ROS. In the present study, the flash-induced thermoluminescence (TL) glow curve parameters and the kinetics of oxygen induction curves of intact leaves and in isolated chloroplasts have been used in order to characterize functional peculiarities of Photosystem II (PSII) in this halophytic plant. TL emission patterns of dark-adapted isolated chloroplasts differs from that of the intact leaves, i.e. B-band (S2QB

- charge pair) peak position occurred at much lower temperatures thus suggesting some solute effects. Illumination of the samples with a series of single turnover flashes resulted in damping of oscillations in B-band intensity as a result of changes in PSII S2QB

- and S3QB- -state cycling. The observed decrease of

the ratio between fast and slow components of oxygen induction curve suggests some changes in the proportion of functionally active PSII alfa and PSII beta centers.

Acknowledgements: This work was partly completed in the frames of bilateral project between Ukrainian Academy of Sciences and Bulgarian Academy of Sciences (2014-2018).


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E3AN OLD BEECH FOREST ON KADIEVI MEADOWS, NORTH PIRIN MT – UNIQUE NATURE WEALTH

ASEN ASENOV¹*, MARIYANA LYUBENOVA²

1 - Department of Botany, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria2 - Department of Ecology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected], Department of Botany, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia 1164, 8 “Dragan Tsankov” Blvd, Bulgaria

Keywords: old beech forest, Pirin Mt, protection

Kadievi Meadows are situated in the foothills of Debeli Hill, North Pirin Mt (2914 m), South-Western Bulgaria. Debeli Hill is the South-Eastern part of North Pirin, connecting it with Middle and Southern Pirin. This part of Pirin is comparatively unknown and rearly visited. The base rock is silicate, the climate is transitional continental and mountain. The Kamenishka river and several streams flow through the meadows, which are used for agriculture and pasture. The highest peak of Debeli Hill is Thcengelchal (Krivets, 2709 m). Four basic cirque with about 20 glacier lakes are grouped along the Debeli Hill. The old park and gallery forest is situated in the Kadievi Meadows locality, on about 1300 m.a.s.l. The groups of very old trees are scattered through the meadows and along the rivers. An old trees have a diameter to 3 m and age over 150-200 years. The beech forest is preserved on the steep slopes. According to Directive 92/43 EEC, they are classificated as 9130 habitat - Asperulo-fagetum. The meadows are classificated as 6520 habitat - Mountain Hay Meadows. That beech forest can accept as national natural heritage, because of the age of the trees. In the last 25 years part of the forest and trees in Kadiwei Meadows were cut. The real danger exists to the other ones. This habitat can be lost. Beech forest of Kadievi Meadows needs attention and protection as soon as it is possible.


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E4ON THE PATTERN OF CORRELATION BETWEEN THE FECUNDITY, LENGTH, WEIGHT AND AGE OF MUGIL CEPHALUS LINNAEUS FROM THE AREAS OF BULGARIAN BLACK SEA COAST

RADOSLAVA BEKOVA1, GALERIDA RAIKOVA-PETROVA2

1 - Department of Marine Geology and Archaeology, Institute of Oceanology, Bulgarian Academy of Science, Varna, Bulgaria2 – Department of General and Applied Hydrobiology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected], 40 First May str., P.O Box 152, 9000 Varna, Bulgaria

Keywords: Mugil cephalus, Black sea coast, fecundity

The paper describes the correlation between the fecundity, length, weight and age of Mugil cephalus Linnaeus from the areas of Bulgarian Black Sea coast. Absolute fecundity varied from a minimum of 423 267 for age 2 (L - 16.9 cm, EW - 51g) to a maximum of 6 002 345 oocytes for age 8 (L - 43.5 cm, EW - 1101 g). The mean fecundity was 305 972 eggs. Relationship between absolute fecundity and weight and fecundity and length was best described by the following equations: F=492043+4216.6W, r=0.9675 and F =-5000000 + 2359954L, r=0.9614. Fecundity-age relationship was best described by the exponential equation: F=77053e1.8797t, r = 0.90. The average weighted relative fecundity was 501. A more accurate indicator to describe the relative fecundity is the coefficient b from the equation witch F=a+bW. For flathead mullet the coefficient b is 4216.6.


90

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E5ASSESSMENT OF ECOSYSTEM SERVICES OF HEATHLAND AND SHRUB ECOSYSTEMS ON THE TERRITORY OF BULGARIA

S. BOTEVA1, R. TZONEV, V. BOGOEV1, K. DIMITROV1, TS. DIMITROVA3, A. KENAROVA1, TS. KOMITOVA3, G. NACHEV2, M. NIKOLOV2, K. PACHEDJIEVA1, N. PARLEVA1, D. SIMEONOVSKA1, V. STEFANOV1, I. TRAYKOV1, R. TSEKOVA1

1 - Sofia University “St. Kliment Ohridski”, Faculty of Biology, Bulgaria, 1164 Sofia, 2 - CARTGEO LTD, Bulgaria, Sofia, 1000,"Vranya" str. 32, floor 3; 3Epsilon Bulgaria. LTD, “Khan Asparuh” 45

Key words: heathland and shrubs ecosystems, ecosystem services, MAES, EU Biodiversity Strategy

The research was carried out during the implementation of the project “Mapping and assessment of heathland and shrub ecosystem (HSE) services in Bulgaria (outside NATURA 2000)”, funded by the Ministry of Environment and Water (MoEW) under the Financial mechanism of the European Economic Area 2009-2014. The wellbeing of humans is integrally linked to the wellbeing of the wildlife, and if the current rate of loss of biological resources is continued, the results will be catastrophic for humankind. The aim of the study is to translate the loss of biodiversity into tangible and quantifiable factors which can be used to promote the development of flexible and effective conservation strategies. This study is relevant to the EU initiative for mapping of the ecosystem services (MAES), which is conducted under action 5 of the EU Biodiversity Strategy. The assessment of ecosystem services follows the methodology of MAES adapted by the MoEW (http://bg03.moew.government.bg/bg). Totally 64 776 plots occupied by HSEs were evaluated to assess the ecosystem services (provisioning, regulating and habitat maintaining, and cultural) provided by them. The areas of HSEs range from 0.25 ha to more than 1041 ha, covering 2.33% of the territory of Bulgaria. Very different is the classification of HSEs depending on the services provided by them. More than 48% of the HSEs are classified as ecosystems, which poorly provide material and energy outputs (provisioning services). The same is the state of cultural services – more than 96% of the HSEs provide poorly to moderately the touristic and religious services. Much better is the state of the regulating (quality of air and soil) services, where around 92% of HSEs provide moderate (62%) to good (30%) state of these services. The SHEs’ service to support the genetic diversity is classified as “very poor”. An IP index (index of ecosystem performance) was calculated for ecosystem conditions and ecosystem services for every plot. Most (89%) of the HSEs are ranked as ecosystems in a good conditions, providing ecosystem services (provisioning, regulating and habitat maintaining, and cultural) in a moderate state. This mismatch indicates that the HSE potential to provide ecosystem services, especially provisioning, is still underestimated by general public and policymakers. The results of our multi-scale study can help to increase awareness of the value of HSE services among stakeholders, improving their knowledge of the functioning of the socio-ecological HSE system, and imposing development of new management strategies for HSE protection.


91

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E6PESTICIDE CONTROL OF ACETYLCHOLINESTERASE ACTIVITY IN SOME INVERTEBRATE ANIMALS AND THE ROLE OF NITRIC OXIDE AS AN ANTIDOTE IN AN ECOTOXICOLOGY ENVIRONMENT

ELITZA T. DENCHEVA1*, RADOY I. IVANOV2

1 - Department of Zoology and anthropology, 2 - Department of Physiology of Animals and Human, Faculty of Biology, Sofia University “St. Kliment Ohridski”,Sofia, Bulgaria*Corresponding author: [email protected], Faculty of Biology, Sofia University “St. Kliment Ohridski”, 8, Dragan Tzankov Blvd., 1164,Sofia, Bulgaria, tel.:+359 2 816 72 35

Keywords: Acetylcholinesterase, invertebrates, L-Arginine, nitric oxide, pesticides, neurotoxicology, antidotes Our previous studies have shown that some donors of nitric oxide (NO) greatly stimulate cholinesterase activity in the nervous tissue of vertebrate animals. The focus of the current study is put on the effect of L-Arginine on the activity of acetylcholinesterase (AChE) in some insect species (Leptinotarsa decemliniata – growth instars and adult forms; Vespulla germanica; Apis mellifera; Gryllus campestris and others). Substantial differences have been established in the base AChE-activity in different species as well as in the larval and adult forms of Leptinotarsa decemliniata. There is a correlative relationship between the activity of this enzyme and the biological, physiological and biochemical characteristics of the respective species.L-Arginine in a concentration of 1.0 to 50 mM and more exercises concentration-dependent stimulating effect on AChE. This effect is dependent on the base level of enzyme activity. The levels of concentration-dependent inhibition of enzyme activity under the influence of carbamate and organophosphate pesticides have also been established. The main result of the study is that L-Arginin and/or NO, L-Citrullin and others, depending on the applied concentration, decrease or eliminate the inhibitory effect of anticholinesterase pesticides and can be used as antidotes against specific poisonings and in an eco-toxic environment.

Acknowledgment: This work was supported by Contract for scientific research № 182/3. 04. 2016 of Sofia University "St. Kliment Ohridski"


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E7DETERMINATION OF TOTAL ANTIOXIDANT CAPACITY IN TWO LYCIUM SPECIES GROWN EX VITRO AT HIGH SALINITY IN HYDROPONIC

VELMIRA DIMITROVA1,2*, TEODORA GEORGIEVA2, MARIA GENEVA3, YULIANA MARKOVSKA1

1 – Department of Plant Physiology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria2 – Biotree LTD, Sofia, 7 Bansko shose str.,Sofia, Bulgaria3 - Department of Plant Soil Interactions, Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Acad. G. Bonchev str., Bl.21, Sofia, Bulgaria*Corresponding author: [email protected], Biotree LTD, Sofia, 7 Bansko shose str.,Sofia, Bulgaria

Keywords: Lycium barbarum, Lycium chinense, K, Na distribution, leaf antioxidant capacity

The effect of salt stress on distribution of alkaline metals in the roots and shoots of Lycium barbarum and Lycium chinense grew ex vitro in hydroponic at three levels of salinity, 50 mM, 100 mM and 200 mM NaCl were investigated. The content of Na and K gradually decreased in the roots and rose in the shoots of Lycium chinense with increasing salinity level as compared to control. The content of Na increased in the roots, but slightly decreased in the leaves of Lycium barbarum. The ratios K/Na were higher in the shoots in comparison with roots of both plants. They were reduced more in the roots of L. barbarum and in the shoots of L. chinense during NaCl treatment. Measured total antioxidant capacity by FRAP assay and DPPH radical method changed by the same manner in the leaves of L. chinense and was enhanced after treatment with 50 mM NaCl, while it gradually decreased in the leaves of L. barbarum. The results were discussed from the view of mechanisms of action of alkaline metals on the generation of reactive oxidant species (ROS) in different organs and the participation of water – and lipid soluble antioxidants in the defense of plants.


93

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E8PHOTOINHIBITION OF PHOTOSYSTEM II RELATED REACTIONS IN THYLAKOID MEMBRANES OF ARABIDOPSIS THALIANA, WT AND MUTANT LUT2, AT ROOM TEMPERATURE

KONSTANTIN DOBREV, GABRIELA PETROVA, DANIELA STANOEVA, MAYA VELICHKOVA, ANTOANETA POPOVA

Bulgarian Academy of Sciences, Sofia, Bulgaria, Institute of Biophysics and Biomedical Engineering, BAS, Acad. Georgi Bonchev Str., Block 21, Sofia 1113Corresponding author: [email protected]

Keywords: photosystem II, photoinhibition, lutein, Arabidopsis thaliana

Carotenoids are intrinsic component of photosynthetic pigment-protein complexes and are involved in performing multiple important functions. Lutein is the most abundant carotenoid in higher plants. The role of lack of lutein in high light induced alterations in structural organization and functional activity of the main pigment-protein complexes was evaluated using isolated thylakoid membranes of Аrabidopsis thaliana, wt and mutant lut2, deficient in lutein. Alterations in photochemical activity of photosystem I and photosystem II were determined by a Clark-type electrode in the presence of exogenous electron donors and acceptors. Activity of oxygen-evolving complex and of the grana and stroma situated photosystem II complexes was evaluated by determination of flash oxygen yields and initial oxygen burst under constant light without donors and acceptors. High light induced alterations in energy transfer and interaction between the main pigment protein complexes was determined by analyzing low-temperature (77K) fluorescence emission and excitation spectra. PAM fluorescence was applied for evaluation of maximal quantum efficiency of PSII (Fv/Fm). Results obtained are discussed in respect to the role of lutein for the organization and sensitivity of photosynthetic apparatus towards high light intensity treatment.

Acknowledgment: This work was supported by Program for career development of young scientists, BAS, under project ДФНП-134/12.05.2016


94

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E9CHANGES OF SUPEROXIDE DISMUTASE ACTIVITY IN THE COMMON BEECH AS A RESPONSE TO HERBIVORY

SIMONA GEORGIEVA1*, PETYA DIMITROVA-MATEVA2, SVETOSLAV ANEV3, NIKOLINA TZVEJKOVA3, MIROSLAVA ZHIPONOVA4, DETELINA PETROVA4, GANKA CHANEVA4

1 – B.Sc. student, University of Forestry, Sofia, Bulgaria2 – South-West State Enterprise - Blagoevgrad, Ministry of Agriculture and Food, Bulgaria3 - Department of Dendrology, University of Forestry, Sofia, Bulgaria4 – Department of Plant Physiology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: superoxide dismutase, Fagus sylvatica, Orchestes fagi

The plant enzymatic antioxidant response seems to be rapidly activated by insect herbivory. Common beech (Fagus sylvatica L.) is one of the most industrially important and highly abundant tree species in central Europe, which is often exposed to insects‘ attacks. Beech weevil (Orchestes fagi L.), in particular, being the most significant pest in beech forests in Bulgaria, causes serious damages and could considerably reduce the yield of wood.The aim of the study was to assess the changes in superoxide dismutase (SOD) activity in the beech trees located at two different altitudes - 680 m asl and 1400 m asl, at the Petrohan Training and Experimental Forest Range, Western Balkan Mountains. We observed that the trees at lower altitude were highly affected by O. fagi infestation. The highest SOD activity in their leaves was measured in the beginning of the vegetation period when the growing larvae had activated the defense systems in the leaf tissues. During the season, SOD activity significantly decreased – it was almost 65% lower in August.

SOD in the leaves of the trees grown at 1400 m asl changed in a different way. The highest enzyme activity was registered in July, which in turn decreased in August.

In general, the lowest values of SOD activities in beech were registered at the end of the vegetation period, especially at lower altitude and that effect could be considered as a result of an overproduction of reactive oxygen species.

Acknowledgment: This work was supported by grant 93/2015 of the University of Forestry, Sofia, Bulgaria


95

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E10COMPARISON OF PHOTOINHIBITION KINETICS OF THYALKOID MEMBARNES ISOLATED FROM CONTROL AND HIGH TEMPERATURE ACCILMATED TOMATO PLANTS

AYGYUN FAIK, MILENA GERGANOVA*, GABRIELA PETROVA, MAYA VELITCHKOVA

Institute of Biophysics and biomedical Engineering, Bulgarian Academy of Sciences,Acad. G. Bonchev str. Bl. 21, Sofia 1113, Bulgaria*Corresponding author: [email protected], Institute of Biophysics and biomedical Engineering, Bulgarian Academy of Sciences,Acad. G. Bonchev str. Bl. 21, Sofia 1113, Bulgaria

Keywords: 77K fluorescence, photoinhibition, photosystem 1, photosystem 2, thylakoid membranes, Solanum licopersicum,

Photoinhibition of thylakoid membranes isolated from control and acclimated to higher temperature tomato plants (Solanum lycopersicum) was investigated. The extent of high light-induced damage of both photosystems was evaluated on the basis of photochemical activity of photosystems in presence of artificial electron acceptor and donors. By using a Clark type oxygen electrode the oxygen evolution (photosystem 2 activity) and oxygen uptake (photosystem 1) were measured. In order to check the alternations in energy distribution and energy interaction between pigment-protein complexes 77K fluorescence emission and excitation spectra were analysed. Data showed that as a results of illumination with white light 1200 µmol.m-2.s-1 the ratio F735/F685 that corresponds to energy distribution between both photosystems increases more considerably in thylakoid membranes from plants acclimated to high temperature for 6 days. The results are discussed in term of the role of acclimation of plants to high temperature for tolerance of photosynthetic apparatus high light stress and the role of pigment protein complexes reorganizations in response to stress.

Acknowledgment: This study is supported by the project №136/12.05


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E11PHOTOINHIBITION IN INDUCED DAMAGES OF PHOTOSYNTHETIC APPARATUS OF TOMATO PLANTS (SOLANUM LYCOERSICUM CV M82)

MILENA GERGANOVA*1, AYGYUN FAIK1, EVA SARVARI2, MAYA VELITCHKOVA1

1 – Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Acad. G. Bonchev str. Bl. 21, 1113 Sofia, BULGARIA2 – Department of Plant Physiology and Molecular Plant Biology, Institute of Biology, Eötvös Loránd University, Pázmány P. sétány 1/C, H-1117 Budapest, Hungary*Corresponding author: [email protected], Institute of Biophysics and biomedical Engineering, Bulgarian Academy of Sciences,Acad. G. Bonchev str. Bl. 21, Sofia 1113, Bulgaria

Keywords: photoinhibition, PAM fluorometry, BN-PAGE electrophoresis, pigment-protein complexes

Climate change intensified the occurrence and severity of abiotic stresses that seriously affect the growth and development of plants. The direct impact of abiotic stress on the activity of photosynthesis in higher plants is disruption of all photosynthesis components such as photosystem I and II, electron transport and ATP generating system. High temperature and high light intensities are the environmental factors affecting many plant cultures. In present study the kinetics of photoinhibition of detached leaves from control and high temperature acclimated tomato were followed in order to reveal how the high temperature-induced alterations of photosynthetic apparatus influenced the response of plants to high light stress. PAM fluorometry was applied for evaluation of photosystem II efficiency and non-photochemical quenching capacity. BN-PAGE was used for analysis of organization of the pigment-protein complexes before and after acclimation. Data presented showed that during acclimation to heat the changes of organization and composition of pigment-protein complexes occurr, that alter the response of leaves to high light stress.

Acknowledgment: This study is supported by the project №136/12.05.2016 оf Program for career development of young scientists, BAS


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E12HEAVY METAL CONCENTRATION IN SEVERAL WETLANDS SITUATED ON THREE BULGARIAN DANUBE TRIBUTARIES

BORISLAVA GYOSHEVA 1*, RADOSTINA HRISTOVA 1, VLADIMIR VALCHEV1

1 – Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences, Sofia, Bulgaria*Corresponding author: [email protected], Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences, 2 Gagarin Street, 1113 Sofia, Bulgaria

Keywords: heavy metals, Danube River wetlands, macrophytes

The heavy metal contents of different aquatic ecosystem components were studied in several wetlands along the Danube River. The studied wetlands included abandoned meanders and sampling stations in the downstream parts of three Danube tributaries. Concentrations of the elements Cu, Pb, Zn, and Cd were determined in water, sediment and plant samples. The study was carried out in April and August 2016. As expected, sediments contained the highest amounts of heavy metals. Water samples contained the lowest concentrations for the investigated elements. Notably, the plant samples from the main macrophyte species were the richest in zinc, followed by copper, lead, and cadmium.

Acknowledgment: This work was supported by grant “Program for career development of young scientists, BAS”.


98

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E13ANALYSIS OF THE EFFICIENCY OF MICROBIAL FUEL CELLS BASED ON SULFATE-REDUCTION PROCESS, INTEGRATED IN ANAEROBIC WETLANDS

ROSEN IVANOV, SVETLANA BRATKOVA, ANATOLIY ANGELOV

Department of Engineering Geoecology, Faculty of Geoexploratio, University of Mining and Geology “St. Ivan Rilski”, Sofia, Bulgaria

Keywords: microbial fuel cells, microbial sulfate-reduction, biofilms

Microbial fuel cells (MFCs) are systems in which microorganisms, using various energy sources (mainly organic compounds, but also inorganic substrates such as hydrogen sulfide or ferrous or ammonium ions) convert this energy directly into electricity. This process is accomplished as the electrons that are detached from the donor at its oxidation are transferred to an insoluble anode of the fuel cell instead to the relevant natural acceptors (oxygen, sulfates, ferric ions, nitrates). In MFCs based on microbial sulfate-reduction process, the electricity generation is related with the reduction of sulfates to hydrogen sulfide, which plays the role of a mediator and on the anodic surface it is oxidized to elemental sulfur. In most cases, at the continuous operational mode of the microbial fuel cell, the elemental sulfur is accumulated on the surface of the electrode or in the area around it, as that leads to a deterioration of the electrochemical characteristics of the cell. In this work three MFCs based on the process of microbial sulfate-reduction were tested for a period of over 18 months, as they were integrated in anaerobic wetlands for mining wastewater treatment. There was established the influence of the formed microbial biofilm consisting of diverse microflora and products from microbial metabolism and from chemical and electrochemical reactions (insoluble sulfides and elemental sulfur) on the efficiency of various components of MFCs.


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E14ACTINOBACTERIA AS A COMPONENT OF MICROBIAL COMMUNITY FORMED ON ROCK PAINTINGS OF MAGURA CAVE, BULGARIA

MILENA MITOVA, MARIA IVANOVA, MIHAIL ILIEV

Department of Microbiology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected], Milena M. Mitova Faculty of Biology, Sofia University, 8 Dragan Tzankov Blvd., 1164 Sofia, Bulgaria

Keywords: biodeterioration, conservation, bat guano, Actinobacteria, Magura cave, rock paintings

The Magura Cave is located in north-western Bulgaria. The cave contains an impressive display of prehistoric paintings made by guano feces from cave dwelling bats. Many different types of microorganisms may grow on such substrates under favorable environmental conditions and have a biodeterioration effect on the paintings. Among numerous biological agents, Actinobacteria play a critically important role in stone deterioration. They cause various types of damage to stone as a result of biofilm formation, physical penetration into the substrate, chemical reaction with the substrate by pigments. he aim of the present study was to investigate the Actinobacteria diversity from samples derived from Gallery with paintings, Magura cave. For this purpose species were identified by their phenotypic and biochemical characteristics. The molecular approach, including the amplification of 16S rDNA was also used. The results showed that the strains are closed to the species of Streptomyces. A better understanding of the microbial diversity present on art objects is important for the development of correct conservation and restoration strategies of the unique rock paintings.Acknowledgment: The study is supported by National Science Fund of Ministry of Education and Science, Bulgaria (Project № ДДВУ/73/2010) and the project 165/04/2014 of Sofia University Science Fund.


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E15ANALYSIS OF VIABILITY THROUGH TETRAZOLIUM TEST OF SEEDS OF TWO ALYSSUM SPECIES OF IMPORTANCE FOR PHYTOMINING FROM SERPENTINES IN ALBANIA

DONALTINA VILA1, KALINA VILA1, DOLJA PAVLOVA2, AIDA BANI1

1 – Agricultural University of Tirana, Faculty of Agronomy and Environment, Agro-Environmental Department, Koder-Kamez, Tirana, Albania2 – University of Sofia, Faculty of Biology, Department of Botany, blvd. Dragan Tzankov 8, 1164 Sofia, Bulgaria*Corresponding author: [email protected], University of Sofia, Faculty of Biology, Department of Botany, blvd. Dragan Tzankov 8, 1164 Sofia, Bulgaria

Keywords: Ni hyperaccumulators, seeds, serpentines, tetrazolium test, viability

The serpentine soils in Albania cover ca. 10% of the territory of the country. These soils are used for phytomining activities as an alternative to local agriculture. The plants suitable for such green technologies are Ni hyperaccumulators, naturally distributed, with good seed production, high success rates of germination, establishment and growth. In order to maximize the yields of the selected ‘metal crop’ experiments on seed viability pattern are of importance. The aims of this study were to evaluate the relation between the viability of the seeds of two metalophyte Alyssum species – A. markgrafii and A. murale with their seed germinability. Two populations of the serpentine endemic and obligate Ni hyperaccumulator A. markgrafii and two populations of the facultative Ni hyperaccumulator A. murale growing in Albania were selected. Tetrazolium test (TZ) for seed viability and an alternative quick method for seed’s germinability were applied. A positive tetrazolium result was obtained if seeds stained light pink as well as red. The germinability of both Alyssum species was significantly low. In general, A. murale demonstrated higher germinability compared to A. markgrafii. The results of the performed test revealed that potential viability is in relation with actual germination percentage calculated.

Acknowledgement: This study was realized within the ERASMUS+ contract between the Faculty of Biology at Sofia University and the Agricultural University of Tirana.


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E16EVALUATION OF SOME WEIHGT INDICES AND SPERM COUNT AFTER HYPOXIC EXPOSURE OF RATS AND MICE

EKATERINA PAVLOVA*, EMILIA PETROVA, YORDANKA GLUHCHEVA, DONIKA DIMOVA, NINA ATANASSOVA

Department of Experimental Morphology, Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences, Sofia, Bulgaria*Corresponding author: [email protected], 1300 Sofia, Acad. G. Bontchev Str, bl. 25, room 330, Bulgaria

Keywords: sodium nitrite, hemic hypoxia, testis, sperm count

Sodium nitrite (NaNO2) is one of the principal food preservatives and colorants in the food industry. Besides the variety of industrial applications, NaNO2 has also some beneficial effects in human and veterinary medicine. However, the over dosage of NaNO2 leads to the formation of methemoglobin in the blood, which does not bind oxygen strongly, thus causing hemic hypoxia. Most mammals have little tolerance to hypoxia and their response involves the activation of regulatory mechanisms at systemic, tissue, and cellular levels. In the testis changes in hormonal profile and vascularisation have been reported. The aim of the present study was to evaluate the early effect of acute hemic hypoxia on some weight indices and sperm count of rats and mice. Adult male rats and mice were intraperitoneally injected with a single toxic dose of NaNO2, 50 mg.kg-1 b.w. and 120 mg.kg-1 b.w., respectively. Treated animals were sacrificed at different time intervals (1h, 5h, 24h and 48h) following the administration. Testes and epididymides were sampled and weighed. Spermatozoa were isolated from both vasa deferentia and counted. In all experimental groups we observed an increase of testis weight/body weight index in both species which is probably due to retention of fluid within the seminiferous tubules. We found statistically significant reduction of the rat sperm count at the 5th and 48th hour after acute hypoxia. Our results in mice showed similar tendency in all investigated periods, although not statistically significant. Future studies would elucidate if hypoxia-induced quantitative changes are associated with histopathological findings in both species.

Acknowledgment: This work is supported by a grant No DMU 03/18 for Young scientists from the Bulgarian National Science Fund.


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E17RELATIONSHIP BETWEEN GROWTH INDEX, SPECIFIC LEAF AREA AND LEAF WEIGHT RATIO OF DECIDUOUS ECOSYSTEMS

SIMONA PETEVA1*, MARIYANA LYUBENOVA1, HRISTINA KALCHEVA2

1 – Department of Ecology and Environmental Protection, Faculty of Biology, Sofia, Bulgaria2 – Institute of Biodiversity and Ecosystem Research, Sofia, Bulgaria* Corresponding author: [email protected]

Keywords: growth index, specific leaf area, leaf weight ratio, meta-analysis, Turkey oak, Hungarian oak, Bulgaria

Forest ecosystems cover about 30 % of the Earth's surface and provide important ecosystem goods and services. Forest traits: specific leaf area (SLA), leaf weight ratio (LWR) and growth index (It) are among the key indicators of forest ecosystems functioning. The investigation deals with meta-analysis (including ANOVA and RDA analysis) for 20 Bulgarian locations of Quercus cerris L. and Q. frainetto Ten. forests. The results are verified with the data from TRY database. For all analysis as a response, variables are chosen selected indexes - SLA, LWR and It and as explanatory variables - listed characteristics of locations: latitude, longitude, altitude, precipitation and temperature. The positive correlation of growth index and SLA and negative - with altitude for all locations are obtained. The growth index correlated negatively with LWR and positively with longitude only for Bulgarian locations. The established dependences is indispensable for higher precision of the ongoing analyzes, filling the gaps of data and creating a new generation of vegetation models.


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E18ALLOMETRIC BIOMASS AND CARBON STOCK ESTIMATION IN SHRUB VEGETATION FOR ASSESSMENT OF ECOSYSTEM SERVICES

FRANCHESKA PETROVA*, VERONIKA DENOVA, KALINA PACHEDJIEVA1, IVAN TRAIKOV1, VLADIMIR STEFANOV1, KRASTIO DIMITROV, IVANA IVANOVA

1 – Department of Ecology and Environmental Protection, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected]

Keywords: shrubs, carbon, allometric equations, ecosystem services

The present study was inspired by the increasing interest in shrub ecosystems in the context of providing important ecosystem services. Apart from the material goods that ecosystems produce they have also important functions such as contributing to climate regulation, reducing air and water pollution, improving human health and well-being, etc. The present study aim to estimate carbon storage of shrubs and shrub ecosystems as a regulating ecosystem service. Biomass is calculated by allometric biomass equations (Blujdaea et. al., 2015) and using the collar diameter height of 10 cm as a main variable. The shrubs from five sample plots (100 m2 each) in shrub ecosystems around Sofia city were measured by digital calipers. Dominant species are Rosa canina, Prunus spinosa, P. cerasifera, Crataegus monogyna. Ecologically these communities are not climax vegetation for the temperate zone and belong to the vegetation class Rhamno-Prunetea. The results for total biomass in shrub species (aboveground dry biomass and belowground biomass) and carbon in shrubs are presented in g/m2. It was established that a shrub ecosystem with 10-15% cover of mixed shrub species accumulates between 150-600 g/m2 dry organic compound depending on the species composition of shtubs and an average amount of 111 g/m2 C. The proportion of herbaceous vegetation (grasses and non-woody plants) is not assessed in this study. Our results are lower but similar to the average of 600 g/m2 aboveground biomass of semi-arid areas in Potugal with 65% cover of shrubs and low trees (Castro et. al., 2009). The average amount of carbon is about 1/3 of the C stock in 5-year black locust plantations in Romania (Ciuvat, 2014). Here, we have illustrated the potential benefits of using data, gathered during summer educational practice in Ecology and Environmental Protection, for the accounting of shrub vegetation carbon stores, for mapping, and managing shrub ecosystems for carbon emission reduction.


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E19ECOLOGICAL POTENTIAL OF MODEL RESERVOIRS IN BULGARIA

MONIKA SUBEVA1, LUBOMIR KENDEROV2,VESELA EVTIMOVA1

1-Institute of Biodiversity and Ecosystem Research, BAS, 1 Tsar Osvoboditel blvd., Sofia 1000, Bulgaria2-Department of General and Applied Hydrobiology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, Bulgaria*Corresponding author: [email protected], Institute of Biodiversity and Ecosystem Research, BAS, 1 Tsar Osvoboditel blvd., Sofia 1000, Bulgaria

Keywords: Water Framework Directive (WFD); physical, chemical and biological parameters; reservoirs, Bulgaria

The Water Framework Directive (WFD) aims to achieve good ecological status (potential for heavily modified water bodies) of surface waters. Ecological status is characterised by the quality, structure and functioning of aquatic ecosystems. It is quantified through chemical and biological quality elements (BEK) for lake ecosystems (natural and artificial), such as phytoplankton, macrophytes, phytobenthos, benthic macroinvertebrate and fish communities. The structure of the zooplankton community may provide valuable information about the state of standing water bodies. Regardless of its great significance in pelagic food web it is missing from the list of biological quality elements in Annex 5 of WFD. Our aim was to study the physical and chemical parameters of the water column, together with biological parameters. Here we present some preliminary results for eight Bulgarian reservoirs: Bebresh, Gorni Dabnik, Telish, Sopot, Sinqta Reka, Konush, Chetiridesette Izvora and Pchelina. Samples were collected from several stations and different horizons in individual seasons – spring and summer 2016.

Acknowledgment: This work was supported by "Updating the typology and qualification system for the assessment of surface water bodies category river, lake and transitional waters during the first RBMP" -2015-2016.


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E20CHANGES IN PHOTOSYNTHETIC ACTIVITY AND FATTY ACID COMPOSITION OF PETASITES HYBRIDUS LEAVES UNDER THE INFLUENCE OF ENVIRONMENTAL FACTORS AT VARIOUS PLANT HABITATS

VESELA YORDANOVA1*, ALBENA IVANOVA2, ALBENA MOMCHILOVA1, SVETLANA MOMCHILOVA3, LILIANA MASLENKOVA1

1 – Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Sofia 1113, Bulgaria2 – Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Sofia 1113, Bulgaria3 - Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, Sofia 1113, Bulgaria*Corresponding author: [email protected], Institute of Biophysics and Biomedical Engineering, Acad. Georgi Bonchev Str., Block 21, Sofia 1113, Bulgaria

Keywords: afterglow band, B-band, fatty acids, Petasites hybridus, Photosystem II, thermoluminescence

Common butterbur (Petasites hybridus (L.) Gaertn., Mey. et Scherb.), Asteraceae family, is a perennial shrub growing most luxuriantly in humid thickets, along the banks of streams, ditches, ravines and rivers, preferably in partial shade, but if there is appropriate amount of water, it can grow in full sun. The rhizomes (Petasitidis rhizoma) and the leaves (Petasitidis folium) are used for years in traditional and modern medicine as a drug. Butterbur extracts are effective in prevention of migrene, astma and allergic rhinits. Main biologically active compounds are secondary metabolites classified as sesquiterpene esters of petasin and furanopetasin. The production of secondary metabolites by plants growing in natural populations is conditioned by environmental factors. Properties of photosynthetic apparatus may contribute to a great extent to plant habitat separation and adaptation to environmental factors, but currently there are no comprehensive studies on the effects of abiotic and phenological factors on growth, photosynthetic activity and accumulation of sesquiterpenes in Petasites. In the present study flash-induced thermoluminescence emission, as indicator of energy balance and functioning of PS2 reaction centers, and changes in fatty acid composition have been used to investigate the adaptive changes in dependence of growth conditions in different Petasites habitats. Petasites leaves were collected from shady and humid places after the flowering period in May – June. Changes in TL B-band parameters, corresponding to S2/3QB-charge pair stabilization, as well as in AG/B ratio, as an indicator of the assimilatory potential are considered to reflect some modification in leaf metabolism as a result of fluctuations in environmental factors at natural habitats. The obtained results may contribute to understanding the physiological background of the variation in secondary metabolites pattern of Petasites.

Acknowledgment: This work was supported by grant № DFNP-131/12.05.2016 financed by Bulgarian Academy of Sciences.

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