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Herein, we report on the venom proteome of Vipera anatolica senliki,a recently discovered and hitherto unexplored subspecies of the critically endangered Anatolian meadow viper endemic to the Antalya Province of Turkey. Integrative venomics, including venom gland transcriptomics as well as complementary bottom-up and top-down proteomics analyses, were applied to fully characterize the venom of V. a. senliki. Furthermore, the classical top-down venomics approach was extended to elucidate the venom proteome by an alternative in-source decay (ISD) proteomics workflow using the reducing matrix 1,5-diaminonaphthalene. Top-down ISD proteomics allows for disulfide bond counting and effective de novo sequencing-based identification of high-molecular-weight venom constituents, both of which are difficult to achieve by commonly established top-down approaches. Venom gland transcriptome analysis identified 96 toxin transcript annotations from 18 toxin families. Relative quantitative snake venomics revealed snake venom metalloproteinases (42.9%) as the most abundant protein family, followed by several less dominant toxin families. Online mass profiling and top-down venomics provide a detailed insight into the venom proteome of V. a. senlikiand facilitate a comparative analysis of venom variability for the closely related subspecies, Vipera anatolica anatolica.

Snake venom variations are a crucial factor to understand the consequences of snakebite envenoming worldwide, and therefore it is important to know about toxin composition alterations between taxa. Palearctic vipers of the genera Vipera, Montivipera, Macrovipera,and Daboiahave high medical impacts across the Old World. One hotspot for their occurrence and diversity is Türkiye, located on the border between continents, but many of their venoms remain still understudied. Here, we present the venom compositions of seven Turkish viper taxa. By complementary mass spectrometry-based bottom-up and top-down workflows, the venom profiles were investigated on proteomics and peptidomics level. This study includes the first venom descriptions of Vipera berus barani, Vipera darevskii, Montivipera bulgardaghica albizona,and Montivipera xanthina, as well as the first snake venomics profiles of Turkish Macrovipera lebetinus obtusa, and Daboia palaestinae, including an in-depth reanalysis of M. bulgardaghica bulgardaghicavenom. Additionally, we identified the modular consensus sequence pEXW(PZ)1–2P(EI)/(KV)PPLE for bradykinin-potentiating peptides in viper venoms. For better insights into variations and potential impacts of medical significance, the venoms were compared against other Palearctic viper proteomes, including the first genus-wide Montiviperavenom comparison. This will help the risk assessment of snakebite envenoming by these vipers and aid in predicting the venoms’ pathophysiology and clinical treatments.

A new subspecies of the Fazila’s Lycian Salamander Lyciasalamandra fazilaeis described based on material from ten localities in the Köyceğiz, Ortaca and Dalaman area in south-western Turkey. It is distinguished from the nominotypical subspecies by differences in the colouration pattern, morphometry and the mitochondrial molecular marker 16S rRNA. The distribution area of the new subspecies is located mainly in the western part of Dalaman River except for two seemingly intermediate populations (Şerefler and Sarsala-Kapıkargın). New localities for the species are reported.