Bayram Göçmen

Mass spectrometry guided venom profiling and bioactivity screening of the Anatolian Meadow Viper, Vipera anatolica.

This contribution reports on the first characterization of the venom proteome and the bioactivity screening of Vipera anatolica, the Anatolian Meadow Viper. The crude venom as well as an isolated dimeric disintegrin showed remarkable cytotoxic activity against glioblastoma cells. Due to the rare occurrence and the small size of this species only little amount of venom was available, which was profiled by means of a combination of bottom-up and top-down mass spectrometry. From this analysis we identified snake venom metalloproteases, cysteine-rich secretory protein isoforms, a metalloprotease inhibitor, several type A2 phospholipases, disintegrins, a snake venom serine protease, a C-type lectin and a Kunitz-type protease inhibitor. Furthermore, we detected several isoforms of above mentioned proteins as well as previously unknown proteins, indicating an extensive complexity of the venom which would have remained undetected with conventional venomic approaches.

Determination of in vivo toxicity and in vitro cytotoxicity of venom from the Cypriot blunt-nosed viper Macrovipera lebetina lebetina and antivenom production

The venomous Levantine viper, Macrovipera lebetina lebetina is endemic to Cyprus. The objective of this study was to investigate in vitro cytotoxicity, in vivo lethality, and antivenom production followed by a re-immunization schedule in mice against Macrovipera lebetina lebetina venom. The LD50 value was estimated as 7.58 mg/kg within 24 hours by different venom doses administrated intraperitoneally in mice. Freunds complete and incomplete adjuvants were used for first and second immunization of mice in antivenom production. A cell-based assay was performed to determine the effects of Macrovipera lebetina lebetina venom and antivenom neutralizing potency on L929 cell viability. The snake venom toxicity and cytotoxicity were examined and comparison of results showed good correlation, the LD50 value was tenfold higher than the IC50 value. The IC50 value was 0.62 ± 0.18 mL after 48 hours treatment while the calculated value was 1.62 ± 0.25 mL for the culture media totally refreshed after two hours treatment with venom. The in vitro efficacy of antivenom against Macrovipera lebetina lebetina venom was found to be low. This is the first report that describes the in vivo and in vitro toxic effects of Macrovipera lebetina lebetina venom and antivenom production against this species.

Viper venom induced inflammation with Montivipera xanthina (Gray, 1849) and the anti-snake venom activities of Artemisia absinthium L. in rat

The present study was conducted to explore the characterization of Montivipera xanthina crude venom partially by in vitro and in vivo and the anti-snake venom activities of Artemisia absinthium L. in comparison with carrageenan-induced acute inflammation model in rats. The LD50 value was estimated as 8.78 mg/kg within 24 h by different venom doses administrated intraperitoneally in mice. The IC50 value was 0.43 ± 0.18 μg/ml after 48 h treatment while the calculated value was 0.73 ± 0.10 μg/ml for the culture media totally refreshed after 2 h treatment with venom. Wistar rats were treated intraperitoneally with A. absinthium extract, 30 min before venom or carrageenan was injected subplantarly into the left hind paw. Intraperitoneal administration of 25 and 50 mg/kg extract was inhibited venom induced paw swelling at 0.5, 1, 2 and 3 h (p < 0.05) while 12.5, 25 and 50 mg/kg extract treatment was inhibited carrageenan-induced paw swelling at 2, 3, 4 and 5 h (p < 0.05). In conclusion, the in vivo toxicity and inflammatory actions and in vitro cytotoxic actions of crude M. xanthina venom were performed as a first report and inhibition of venom-induced inflammation by methanolic extract of A. absinthium was described.

Ciliated Protozoa in the Rumen of Turkish Domestic Cattle (Bos taurus L.)

Abstract Rumen contents obtained from 28 domesticated cattle (Bos taurus L.) slaughtered at abatoirs near Izmir, Turkey were surveyed for ciliate protozoa. Protozoa are known to make an appreciable contribution to ruminal fiber digestion in many different geographical areas; however, little if any information is available on their occurrence in Turkish cattle. As a result of our survey, 13 genera including 52 species were identified. Nine of the species were further divided into 36 forma. The average ciliate density in our cattle (52.44 × 104/ml) was higher than that of Turkish domestic sheep and other domestic cattle reported previously from different geographical areas. Entodinium basoglui, Entodinium williamsi f. williamsi, E. williamsi f. turcicum, E. dalli f. rudidorsospinatum, Entodinium imai, Entodinium oektemae, Eudiplodinium dehorityi, Epidinium graini, Ophryoscolex purkynjei f. bifidobicinctus, and Ophryoscolex purkynjei f. bifidoquadricinctus have previously been reported from Turkey and appear to be endemic. All of the remaining species represent a new host record for domestic Turkish cattle. This study also reports for the second time the presence of Entodinium constrictum in herbivorous mammals, and is the first record of its occurrence in domesticated cattle.

The Rumen Ciliate Fauna of Domestic Sheep (Ovis ammon aires) from the Turkish Republic of Northern Cyprus

Abstract Concentration and composition of ciliate protozoa in the families Ophryoscolecidae and Isotrichidae were determined in rumen contents of domestic sheep (Ovis ammon aries) from Cyprus. A total of five genera of Ophryoscolecidae were identified, Metadinium, Enoploplastron, Polyplastron, Epidinium, and Ophryoscolex, which included six species: Metadinium affine, Enoploplastron triloricatum, Polyplastron multivesiculatum, Epidinium ecaudatum, Epidinium graini, and Ophryoscolex purkynjei. Eight separate forms of Epidinium were identified (E. ecaudatum f. ecaudatum, E. e. f. caudatum, E. e. f. bicaudatum, E. e. f. tricaudatum, E. e. f. quadricaudatum, E. graini f. graini, E. g. f. caudatricoronatum, and E. g. f. caudaquadricoronatum), along with five forms of Ophryoscolex purkynjei (O. p. f. purkynjei, O. p. f. bifidobicinctus, O. p. f. bifidoquadricinctus, O. p. f. bicoronatus, O. p. f. tricoronatus, and O. p. f. quadricoronatus). Three species of Isotrichidae were observed, Isotricha intestinalis, I. prostoma, and Dasytricha ruminantium. This study reports new host records for three forms of Epidinium graini and Ophryoscolex purkynjei f. bifidobicinctus. The rumen fauna in the family Ophryoscolecidae from Cypriote domestic sheep appear to have limited diversity compared to those from Turkish and Far Eastern (Chinese/ Japanese) sheep, while they are more diverse than those found in Western European (Scottish) and North American (Canadian/Alaskan) sheep.

Redescription of Triplumaria selenica Latteur et al., 1970 (Ciliophora, Entodiniomorphida) and its phylogenetic position based on the infraciliary bands and 18SSU rRNA gene sequence

Triplumaria selenicaLatteur, Tuffrau and Wespes, 1970 was redescribed from pyridinated silver carbonate-impregnated specimens. Triplumaria selenica has a slit of the vestibular opening extending posteriorly along the left side of the vestibulum. The wide C-shaped adoral polybrachykinety extends along the ventral side of the vestibular opening. The narrow perivestibular polybrachykinety extends laterally along the dorsal side of the vestibular opening from the right end of the adoral polybrachykinety and forms a loop extending posteriorly along the vestibular slit to join to the left end of the adoral polybrachykinety. The 18SSU rRNA gene of T. selenica as well as those of six other entodiniomorphid species, Raabena bella, Blepharocorys curvigula, Entodinium longinucleatum, Eudiplodinium rostratum, Metadinium medium, and Ostracodinium gracile was sequenced. The neighbor joining and maximum parsimony phylogenetic trees were constructed to discuss the evolution of entodiniomorphs. Our results will support and extend Wolskas hypothesis: the ancestral forms of blepharocorythids have evolved into ophryoscolecids and Cycloposthium species via the ancestor of Triplumaria.

Taxonomic notes on the snakes of Northern Cyprus, with observations on their morphologies and ecologies

A total of 66 specimens belonging to eleven snake species, some of which were collected during the fi eld studies of 17-31 July and 3-25 September 2003, and some others which have been previously collected and all have been deposited into the collection of ZDEU (Zoology Dept. Ege University), were evaluated taxonomically. 25 of the specimens belonged to Typhlopidae, 37 to Colubridae, three to Viperidae and one to Boidae. Th ese specimens were evaluated from the points of view of pholidosis, pattern and coloration and morphological measurements. Some biological and ecological information were also given with the brief geological history of eastern Mediterranean region.

New rumen ciliates from Turkish domestic cattle (Bos taurus L.) I. The presence of Entodinium dalli Dehority, 1974 with a new forma, E dalli f. rudidorsospinatum n. f. and comparisons with Entodinium williamsi n. sp.

Summary In the course of examining rumen contents obtained from 25 domestic cattle ( Bos taurus ) in the vicinity of Izmir, Turkey, two somewhat similar but unusual forms of entodinia were observed. These ciliates were observed in three of the animals and this study reports on their classification into two species, Entodinium dalli Dehority, 1974 and a new species, Entodinium williamsi n. sp. All the protozoa identified as belonging to E. dalli were distinguished as a new forma, E. dalli f. rudidorsospinatum n. f. of the basis of their caudal spines. The species Entodinium williamsi n. sp. was also divided into two formae, E. williamsi f. williamsi n. f. and E. williamsi f. turcicum n. f. The morphological characteristics of these protozoa are described and their relationships to similar entodiniid species are discussed.

Histochemical and Biometric Study of the Gastrointestinal System of Hyla Orientalis (Bedriaga, 1890) (Anura, Hylidae)

This study was carried out to assess the localization of hyaluronic acid (HA) and the distribution of glycoproteins in the gastrointestinal system of adult Hyla orientalis. Histochemical analysis of the gastrointestinal system in H. orientalis showed that mucous content included glycogene and/or oxidable dioles [periodic acid/Schiff (PAS)+], neutral or acid-rich (PAS/AB pH 2.5+), sialic acid residues (KOH/PAS+) and acid sulphate [Aldehyde fuchsin (AF)+] glycoproteins. However the mucus content was not the same in stomach, small and large intestine. The mucus content of stomach included only glycogene and/or oxidable dioles and sialic acid residues. Besides these histochemical methods, the localization of HA was detected using biotinylated hyaluronic acid binding protein labeled with streptavidin-fluorescein isothiocyanate (FITC). In the extracellular matrix of the submucosa, the reaction for HA was evident. Since HA was located in submucosa beneath the epithelial layer of gastrointestinal system, it has a significant role in hydric balance, and essential to provide the gastrointestinal system integrity and functionality. According to biometric results, there were statistical differences between small and large intestine in terms of the amount of material stained positive with PAS/AB, PAS, KOH/PAS and AF/AB. Additionally, number of goblet cells in the small and large intestine was significantly different.

Morphological and serological investigations on Lacerta laevis Gray, 1838 (Sauria: Lacertidae) populations from Anatolia

Abstract A total of 51 specimens of Lacerta laevis (25 ♂, 26 ♀) from the area of Mersin and Hatay (southern Anatolia) were investigated morphologically, and of these 24 specimens (12 ♂, 12 ♀) were analyzed from the viewpoint of their blood-serum proteins. The populations were found to show great intra- and interpopulation variation in their pattern and colour characteristics and in their serological analyses, while their morphometric characteristics were found to be similar. It is thus concluded that, for the present, it is not possible to separate them into different subspecies.