DNA, spectroscopic and geochemical analyses of bone fragments and associated speleothems in Postojna cave, Slovenia

a loose/broken stalagmite containing small fragments of cemented bones were collected from the Postojna Cave to investigate whether deoxyribonucleic acid (DNA) can be determined. The study is complemented by the Fourier-transform infrared spectroscopy - attenuated total reflectance (FTIR-ATR) analysis in order to determine the alteration of the bones and to test whether this analysis can be used as an indicator of possible DNA preservation. In addition, geochemical analyses were conducted in order to determine whether the associated flowstone/stalagmite is suitable for elucidating the timing of bone thanatocoenosis and further palaeoenvironmental analyses. The organic matter (collagen) is poorly preserved. However, we succeeded in amplifying a 94 bp long fragment of the cytochrome b (Cyt b) gene of mitochondrial DNA (mtDNA) in polymerase chain reaction (PCR) for one sample, and in sequencing the amto that of the Cyt b of the cave bear (Ursus deningeri or Ursus spelaeus sensu lato). The uranium-thorium dating of the speleothem covering the bones revealed its thanatocoenosis occurred prior to 55 ka, most likely in the late marine isotope stage 4 or early marine isotope stage 3. High porosity and recrystallisation of the flowstone/stalagmite at this part of the cave prevent high-resolution palaeoclimatic interpretation; however, low-resolution stable isotope geochemistry suggests a steppe-like environment during the subsequent growth of the speleothem.Iz Postojnske jame so bili vzeti delci kosti, prevlečeni s kalcitno skorjo, vključno z odlomljenim stalagmitom, v katerem so zacementirani drobci kosti, da bi se ugotovilo, ali je mogoče določiti deoksiribonukleinsko kislino (DNA). Študijo dopolnjuje analiza s Fourierevo transformacijsko infrardečo spektroskopijo z zmanjšanim popolnim odbojem (FTIR-ATR), ki je bila izvedena, da se ugotovijo biokemične spremembe v kosteh in se preveri, ali je mogoče to analizo uporabiti kot kazalnik morebitne ohranjenosti DNA. Izvedene so bile tudi geokemijske analize, da bi ugotovili, ali je pripadajoča siga primerna za razlago časa kostne tanatocenoze in nadaljnje paleookoljske analize. Organska snov (kolagen) je slabo ohranjena. Pri enem vzorcu nam je z verižno reakcijo s polimerazo (PCR) uspelo pomnožiti 94 bp dolg fragment gena za citokrom b (Cyt b) mitohondrijske DNA (mtDNA) in pomnožene produkte smo uspešno sekvencirali. Dobljeno zaporedje dolžine 70 bp ustreza zaporedju Cytb jamskega medveda (Ursus deningeri ali Ursus spelaeus sensu lato). U-Th datacija sige, ki pokriva kosti, nakazuje, da se je tanatocenoza pojavila pred 55 ka, najverjetneje v fazi MIS 4 ali MIS 3. Visoka poroznost in rekristalizacija sige v tem delu jame preprečujeta paleoklimatsko interpretacijo visoke ločljivosti, geokemija stabilnih izotopov z nizko ločljivostjo pa nakazuje na stepi podobno okolje

Prions and animal transmissible spongiform encephalopathies

Background. Transmissible spongiform encephalopathies (TSEs) or prion diseases are a unique group of neurodegenerative diseases of animals and humans, which always have a fatal outcome and are transmissible among animals of the same or different species. Scope and Approach. The aim of this work is to review some recent data about animal TSEs, with the emphasis on their causative agents and zoonotic potential, and to discuss why the surveillance and control measures over animal TSEs should remain in force. Key Findings and Conclusions. We still have incomplete knowledge of prions and prion diseases. Scrapie has been present for a very long time and controlled with varied success. Bovine spongiform encephalopathy (BSE) emerged unnoticed, and spread within a few years to epidemic proportions, entailing enormous economic consequences and public concerns. Currently, the classical BSE epidemic is under control, but atypical cases do, and probably will, persist in bovine populations. The Chronic Wasting Disease (CWD) of the cervids has been spreading in North America and has recently been detected in Europe. Preventive measures for the control of classical BSE remain in force, including the feed ban and removal of specified risk materials. However, active BSE surveillance has considerably decreased. In the absence of such preventive and control measures, atypical BSE cases in healthy slaughtered bovines might persist in the human food chain, and BSE prions might resurface. Moreover, other prion strains might emerge and spread undetected if the appropriate preventive and surveillance measures were to cease, leaving behind inestimable consequences

Comparison of mtDNA control region among descendant breeds of the extinct Zaupel sheep revealed haplogroup C and D in Central Europe

The consideration of the descendance is indispensable in the preservation of endangered animal breeds. The authors compared mitochondrial DNA (mtDNA) control region sequence in three descendant breeds of the extinct Zaupel sheep, firstly. Their investigation was carried out in order to prove the common origin of Waldschaf (Austria), Bovec sheep (Slovenia) and Cikta (Hungary). A total of 118 biological samples were taken from non-related representatives of the three breeds between 2015 and 2017. A newly designed primer pair was also used to amplify the segment (1180 bp) to be tested. The total number of haplotypes in the whole study population was 49. The majority of which fell into haplogroup B. The significant negative value of the Fu\u27s Fs statistic (Fs statistic = −3.296, p = 0.013) based on haplotype frequencies demonstrated a moderate foreign gene flow. As a novel observation haplogroups C and D appeared in Cikta and Bovec sheep, respectively. The Tajima D-test value in the entire study population was −0.914 (p > 0.10), meaning that the separation of the three descendant breeds did not cause genetic drift, these are collectively in genetic equilibrium. The genetic information confirmed the common origin of the breeds known from the breed history