Human gastrointestinal parasites are good indicators for hygienic conditions and health

Human gastrointestinal parasites are good indicators for hygienic conditions and health status of past and present individuals and communities. 100 to 7 200 year-old archeological samples proved this to be a powerful reliable and efficient approach for species determination even in the absence of preserved eggs either as a stand-alone method or as a complement to microscopic research. Introduction Attacks with gastrointestinal helminths certainly are a main public wellness concern. Soil-transmitted helminthiases influence about 2 billion people world-wide [1 2 Quotes claim that the nematode infects almost 1.2 billion people and 800 million. attacks due to ingesting food or soil contaminated with their eggs result in roughly 60 0 deaths per year [1 2 The eggs of the tapeworms (Cestoda) contamination can cause life-threatening cysticercoses. The flukes and approach to the analysis of ancient parasite DNA preserved in archaeological sediments of various origins (such as latrines cesspits human burials etc). The approach is based on multiplex PCR optimized for highly degraded ancient DNA molecules followed by next generation sequencing of the amplicons around the Ion Torrent platform to efficiently and inexpensively genotype large numbers of archaeological samples [13]. We designed PCR primers to target different taxa of human parasites that are commonly found in archeological sites taking into account both the short fragment length of ancient DNA molecules and the genetic diversity of the taxa analyzed. Primers were optimized within four multiplex PCRs to allow the identification of 16 species of human tapeworms roundworms pinworms and flukes in up to 96 samples simultaneously. The results obtained with this approach on a variety of ca. 100 to 7 200 year-old samples from numerous archeological and taphonomic contexts were compared with those obtained with the microscopic approach. This study reveals the power of the genetic approach and reports the detection of parasite DNA even in the absence of corresponding eggs. This new approach GSK1120212 for the genetic identification of parasites in archeological sites enables researchers to trace back parasite lineages and better understand their development. Additionally the higher resolution analyses possible through genotyping can clarify transmission events from populace migrations and contacts between populations the conquest of new environments demographic changes and the domestication of animals during the Neolithic and overall health status of past individuals and populations. Materials and Methods No permits were required for the explained study in particular ground sampling which complied with all relevant regulations. Archeological samples Sediments GSK1120212 from archeological sites have been collected from a wide variety of archeological and taphonomic contexts geographic areas and periods dating from your Iberian Neolithic (ca. 7 200 BP) to the last century (First World War) (observe Table 1). Table 1 List of samples analyzed with results of both genetic (DNA sequences) and microscopic analyses (Eggs). Microscopic analysis Prior to genetic analysis all samples have been analyzed according to standard paleoparasitological protocols at the laboratory of paleoparasitology of the University or college of Franche-Comté [14]. For each sample five grams of sample were rehydrated for one week in a mixed answer of 0.5% tri-sodium phosphate (TSP) and 5% glycerinated water. The samples were then crushed in GSK1120212 a mortar and subjected to an ultrasound bath for one minute before finally being filtered in a column composed of sieves with 315 160 50 and 25 μm meshes. Since eggs varied in size between 30-160 μm in length and 15-90 μm in width [15] residues from the two last sieves (50 and 25 μm) were transferred to PVC tubes. Ten slides (22×22 mm) were prepared from each portion and analyzed under the light Rabbit Polyclonal to FES. microscope amounting to GSK1120212 about 5% of the recovered fraction. Extraction and purification of ancient DNA The pre-PCR GSK1120212 procedures were carried out in the high containment historic DNA analysis service with positive surroundings pressure from the Jacques Monod Institute using tight experimental techniques as defined [16]. Two to ten grams of sediment had been ground to great powder utilizing a Fridge Mill (6750 Spex Certiprep Metuchen NJ). The powder was purified using PowerMax? Garden soil DNA Isolation Package (MO BIO Laboratories Inc. Carlsbad CA) pursuing.